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The regulation of the cell cycle is essential for maintaining normal cellular function, especially in the development of diseases such as lung cancer. The cell cycle consists of four major phases (G1, S, G2 and M phases), which are characterized by a series of precise molecular events to ensure proper cell proliferation and division. In lung cancer cells, cell cycle dysregulation can lead to disordered proliferation and increased invasiveness of cancer cells. G2 and S-phase expressed 1 (GTSE1) is a regulatory protein found in the cytoplasm of the cell, which plays a key role in the cell cycle distribution of a wide range of cancer cells and is involved in life processes such as cell proliferation and apoptosis. GTSE1 affects cell cycle progression by interacting with cyclin-dependent kinase inhibitor 1A (p21) and maintaining the stability of p21, which in turn inhibits the activity of cyclin-dependent kinase 1/2 (CDK1/2). In addition, GTSE1 is also involved in the regulation of tumor protein 53 (p53) signaling pathway. With the assistance of mouse double minute 2 homolog (MDM2), GTSE1 is able to transport p53 from the nucleus to the cytoplasm and promote its ubiquitination and degradation, thus affecting cell cycle and cell deathrelated signaling pathways. This paper reviews the expression of GTSE1 in lung cancer cells and its effects on lung cancer, as well as its potential mechanisms involved in cell cycle regulation. [ABSTRACT FROM AUTHOR]

Background and purpose: Lung adenocarcinoma has the characteristics of difficult early detection, rapid tumor progression and low surgical resection rate. Although studies on immunotherapy alone and immunotherapy combined with chemotherapy have shown initial success in improving prognosis and overcoming drug resistance, the majority of lung adenocarcinoma patients still receive limited benefits. Therefore, there is an urgent need to identify novel biomarkers with relatively high sensitivity and specificity to improve the prognosis of lung adenocarcinoma. Cell division cycle protein 20 (CDC20) is involved in the occurrence and development of various tumors, but its biological role and mechanism in lung adenocarcinoma remain unclear. The aim of this study was to investigate the expression of CDC20 in lung adenocarcinoma and its predictive value for the prognosis of patients with lung adenocarcinoma, and to further explore the effects of CDC20 on the proliferation and invasion capabilities of lung adenocarcinoma cells. Methods: Utilizing immunohistochemistry (IHC) to detect the expression of CDC20 in lung adenocarcinoma tissues, we analyzed its correlation with poor prognosis in combination with bioinformatics and clinicopathological parameters. Kaplan-Meier survival curves were employed to illustrate the impact of CDC20 on postoperative survival rates in lung adenocarcinoma patients. COX multivariate regression analysis was conducted to identify independent prognostic factors influencing postoperative survival rates. Additionally, receiver operating characteristic (ROC) curves were applied to assess the diagnostic value of CDC20 expression in lung adenocarcinoma patients. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were used to measure the expression levels of CDC20 in normal human lung epithelial cells (BEAS-2B) and human lung adenocarcinoma cells (A549 and H1299). In cellular experiments, CDC20 was knocked down in lung adenocarcinoma cells, which were divided into three groups: si-NC (control group), si-CDC20#1 (knockdown group 1) and si-CDC20#2 (knockdown group 2). Cell counting kit-8 (CCK-8), colony formation, transwell and wound healing assays were conducted to assess cell proliferation, migration and invasion capabilities. Functional enrichment analysis using Geng Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways was conducted to explore the biological roles of CDC20 in lung adenocarcinoma. Finally, gene set enrichment analysis (GSEA) was employed to investigate potential regulatory pathways of CDC20 in lung adenocarcinoma. This study was approved by the Ethics Committee of Hebei Chest Hospital (Number: 2022051). Results: The results of both bioinformatics analysis and IHC demonstrated a significantly high expression of CDC20 in lung adenocarcinoma tissues (P 0.05). Both bioinformatics analysis and clinical parameter evaluation revealed a correlation between high CDC20 expression and poor patient prognosis. Kaplan-Meier survival analysis and COX regression analysis consistently indicated a significant negative correlation between CDC20 expression and postoperative survival rates in patients (P0.05). Additionally, the expression levels of CDC20 were higher in human lung adenocarcinoma cell lines A549 and H1299 compared with BEAS-2B (P0.05). Knockdown of CDC20 effectively inhibited the proliferation, migration and invasion of lung adenocarcinoma cells (P0.05). The results of GO, KEGG pathways and GSEA consistently pointed to a relationship between CDC20 and cell cycle regulation. Conclusion: CDC20 is highly expressed in lung adenocarcinoma. High expression of CDC20 is an independent risk factor for poor prognosis of lung adenocarcinoma patient. CDC20 can promote the proliferation, migration and invasion of lung adenocarcinoma cells. [ABSTRACT FROM AUTHOR]

Background and purpose: Esophageal carcinoma (ESCA) is one of the malignant tumors with high mortality rate, and the underlying mechanism of its development is largely unknown. CDC20 plays an important role in tumorigenesis, and its dysregulated expression is closely related to tumor occurrence and development. The expression of CDC20 is increased in a variety of tumors, and knocking down CDC20 can inhibit tumor cell proliferation. NLRP3 is the main component of the inflammasome, and inflammasome is also closely related to tumor occurrence and development. Here, our study aimed to investigate whether CDC20 promotes the proliferation of ESCA cells through NLRP3 and its regulatory mechanism. Methods: The expression levels of CDC20 and NLRP3 genes in ESCA patients were analyzed using The Cancer Genome Atlas (TCGA) detabase and GTEx public database. We collected clinical and pathological data and tissues from 80 ESCA patients at the First Affiliated Hospital of Xinxiang Medical College, and detected the protein expression of NLRP3 in ESCA patients through immunohistochemistry staining. This study was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical College (Number: EC-021-137). We studied the effects of knocking down CDC20 and NLRP3 gene on the proliferation ability of esophageal squamous cell carcinoma cells EC9706 and KYSE150 using short hairpin RNA (shRNA) technology. Co-immunoprecipitation (Co-IP), proteasome inhibitors and ubiquitination experiments were used to detect whether CDC20 interacts with NLRP3, and to elucidate whether CDC20 regulates NLRP3 expression through the ubiquitination pathway. This study was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical College (Number: EC-021-137). Results: The TCGA database analysis showed that the expression levels of CDC20 and NLRP3 mRNA were significantly higher in the cancer tissues of ESCA patients than in the adjacent tissues. The immunohistochemistry results further showed that compared with adjacent tissues, the protein expression levels of CDC20 and NLRP3 were increased in ESCA tissues. Knocking down CDC20 and NLRP3 genes inhibited the proliferation of ESCA cells. Co-IP, proteasome inhibitors and ubiquitination experiments confirmed that CDC20 interacted with NLRP3 through its leucine-rich repeat (LRR), and CDC20 stabilized its expression by promoting NLRP3 ubiquitination. Conclusion: CDC20 and NLRP3 are upregulated in ESCA tissues, and CDC20 stabilizes their expression through ubiquitination of NLRP3, promoting ESCA cell proliferation. This suggests that CDC20 and NLRP3 may be potential diagnostic targets for ESCA. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies have found that qi deficiency and blood stasis syndrome is the main syndrome among various TCM syndromes of cervical spondylotic myelopathy. However, there is no report on proteomic markers as early diagnosis indicators for the transformation of developmental cervical spinal stenosis with qi deficiency and blood stasis syndrome to cervical spondylotic myelopathy. OBJECTIVE: To explore serum proteomics difference between developmental cervical spinal stenosis and cervical spondylotic myelopathy and to find and identify the potential serum biomarkers between them. METHODS: Serum samples of nine patients with cervical spondylotic myelopathy of qi deficiency and blood stasis syndrome (experimental group) and nine patients with developmental cervical spinal stenosis of qi deficiency and blood stasis syndrome (control group) were collected. The proteomic analysis was carried out by Tandem Mass Tag combined with liquid chromatography tandem mass spectrometry, so as to find and identify differentially expressed proteins. RESULTS AND CONCLUSION: A total of 1027 significantly differential proteins were initially screened by TMT technology and 89 significantly differential proteins were finally identified (P < 0.05). Compared with the control group, there were 45 up-regulated proteins in the experimental group, such as α-actinin-4, α-actinin-1, cell division control protein 42 homolog, integrin-linked protein kinase and B-actin. Conversely, there were 44 down-regulated proteins in the experimental group compared with the control group, such as fibronectin, fibrinogen γ chain, fibrinogen α chain, fibrinogen β chain. Gene ontology enrichment analysis indicated that these differential proteins were involved in signal receptor binding, kinase binding, protein kinase activity, integrin binding, actin filament binding and other molecular functions. Based on the Kyoto Encyclopedia of Genes and Genomes pathway analysis, 20 common differential signal/metabolic pathways were identified, including Rap1 signaling pathway, adherens junction, tight junction, platelet activation, and regulation of actin cytoskeleton. Proteinprotein interaction analysis showed that ILK, FGA, FGB, FGG, FN1, Cdc42, ACTN1, ACTN4 and ACTB were located at the nodes of protein-protein interaction network and were closely related to bone formation and destruction system, nervous system, coagulation system, cellular inflammation and other systems. To conclude, the serum differentially expressed proteins between developmental cervical spinal stenosis and cervical spondylotic myelopathy can be successfully screened by Tandem Mass Tag combined with liquid chromatography tandem mass spectrometry. ILK, FN1, CDC42 and ACTN 4 are identified as specific markers for the transformation of developmental cervical spinal stenosis with qi deficiency and blood stasis syndrome into cervical spondylotic myelopathy. These findings provide a basis for further clarifying the transformation mechanism. [ABSTRACT FROM AUTHOR]

Background and objective Radiation therapy is one of the most common treatments for non-small cell lung cancer (NSCLC). However, the insensitivity of some tumor cells to radiation is one of the major reasons for the poor efficacy of radiotherapy and the poor prognosis of patients, and exploring the underlying mechanisms behind radioresistance is the key to solving this clinical challenge. This study aimed to identify the molecules associated with radioresistance in lung adenocarcinoma (LUAD), identified thyroid hormone receptor interactor 13 (TRIP13) as the main target initially, and explored whether TRIP13 is related to radioresistance in LUAD and the specific mechanism, with the aim of providing theoretical basis and potential targets for the combination therapy of LUAD patients receiving radiotherapy in the clinic. Methods Three data-sets, GSE18842, GSE19188 and GSE33532, were selected from the Gene Expression Omnibus (GEO) database and screened for differentially expressed genes (/log FC/>1.5, P<0.05) in each of the three datasets using the R 4.1.3 software, and then Venn diagram was used to find out the differentially expressed genes common to the three datasets. The screened differential genes were then subjected to protein-protein interaction (PPI) analysis and module analysis with the help of STRING online tool and Cytoscape software, and survival prognosis analysis was performed for each gene with the help of Kaplan-Meier Plotter database, and the TRIP13 gene was identified as the main molecule for subsequent studies. Subsequently, the human LUAD cell line H292 was irradiated with multiple X-rays using a sub-lethal dose irradiation method to construct a radioresistant cell line, H292DR. The radioresistance of H292DR cells was verified using cell counting kit-8 (CCK-8) assay and clone formation assay. The expression levels of TRIP13 in H292 and H292DR cells were measured by Western blot. Small interfering RNA (siRNA) was used to silence the expression of TRIP13 in H292DR cells and Western blot assay was performed. The clone formation ability and migration ability of H292DR cells were observed after TRIP13 silencing, followed by the detection of changes in the expression levels of proteins closely related to homologous recombination, such as ataxia telangiectasia mutated (ATM) protein. Results Screening of multiple GEO datasets, validation of external datasets and survival analysis revealed that TRIP13 was highly expressed in LUAD and was associated with poor prognosis in LUAD patients who had received radiation therapy. And the results of gene set enrichment analysis (GSEA) of TRIP13 suggested that TRIP13 might be closely associated with LUAD radioresistance by promoting homologous recombination repair after radiation therapy. Experimentally, TRIP13 expression was found to be upregulated in H292DR, and silencing of TRIP13 was able to increase the sensitivity of H292DR cells to radiation. Conclusion TRIP13 is associated with poor prognosis in LUAD patients treated with radiation, possibly by promoting a homologous recombination repair pathway to mediate resistance of LUAD cells to radiation. [ABSTRACT FROM AUTHOR]

Objective To identify the key genes associated with the early diagnosis and poor prognosis of hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) by using the bioinformatics methods, and to elucidate the underlying molecular mechanism of occurence and development of HBV-HCC. Methods Gene Expression Omnibus (GEO) Database was used to retrieval “hepatitis B induced HCC”； the Gene Dataset GSE121248 was downloaded, the differentially expressed genes (DEGs) were screened by the “limma” Data Package in R Software, and the DEGs were enriched by using the “clusterProfiler” Data Package for Gene Ontology (GO) functional analysis and the Kyoto Genes and Genome Encyclopedia (KEGG) signaling pathway enrichment analysis； STRING Database and Cytoscape Software were used to establish the protein-protein interaction (PPI) network and the key genes were screened out.Gene Expression Profiling Interactive Analysis (GEPIA), Kaplan Meier-Plotter，and Human Protein Atlas (HPA) Databases were used to verify the key genes and the expression levels of proteins；the infiltration of the immune cells was analyzed based on the “CIBERSORT” Data Package. Results A total of 574 DEGs were identified，including 173 up-regulated genes and 401 down-regulated genes. The GO functional enrichment analysis results showed that DEGs were mainly enriched in the biological processes such as small molecule metabolism, signal transduction, immune response, inflammatory response，and so on； the KEGG signaling pathway enrichment analysis results showed that the DEGs were mainly enriched in retinol metabolism, cytochrome P450 metabolic pathway of exogenous drugs, and chemical carcinogenesis，and so on. The PPI network results showed that cell division cycle 20 (CDC20) ，cyclin dependent kinase 1 (CDK1), cyclin A2 (CCNA2), pindle checkpoint protein (BUB1B), topoisomerase Ⅱ α (TOP2A), discs large homolog associated related protein 5 (DLGAP5), abnormal spindle-like microcephaly associated protein (ASPM), centrosomal protein 55 (CEP55), kinesin superfamily 11 (KIF11) ，and kinesin superfamily 20A (KIF20A) were the key genes. The GEPIA Database analysis results showed that these above 10 key genes were highly expressed in the HCC patients. The Kaplan Meier survival curve showed that the overall survivals of the HCC patients with high expression of key genes were shorter than those of the HCC patients with low expression of key genes. Conclusion The genes related to cell cycle and viral oncogenesis (CDC20, CDK1, CCNA2, and BUB1B) are closely associated with the occurence and development and poor prognosis of the HBV-HCC patients, which may become the diagnostic markers and new targets for the treatment. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To detect the expression levels of serum and glucocorticoid-induced protein kinase 3 (SGK3) in one-cell stage fertilized eggs at different cell cycles of the mice and its subcellular localization, and to preliminarily clarify the regulatory effect of SGK3 on the early development of fertilized eggs of the mammal. Methods: The one-cell stage fertilized eggs of the mice were collected by superovulation technique and in vitro culture of fertilized eggs, the expression levels of SGK3 mRNA and protein in the one-cell stage fertilized eggs at different cell cycles were detected by real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting methods, respectively; the subcellular localization of SGK3 in the one-cell stage fertilized eggs at different cell cycles of the mice was observed by immunofluorescence assay. Results: The SGK3 mRNA and protein were expressed in the one-cell stage fertilized eggs at different cell cycles of the mice; compared with G1, S and M phases, the expression levels of SGK3 mRNA and protein in one-cell phase fertilized eggs at G2 phase were increased (P<0. 01). The immunofluorescence results showed that SGK3 (red fluorescence) was localized in cytoplasm of the one-cell phase fertilized eggs at G1 and S phases, and some SGK3 entered into the nucleus at G2 phase, and the SGK3 was widely distributed throughout the cells at M phase. Conclusion: The SGK3 in the one-cell stage fertilized eggs at different cell cycles of the mice dynamically expresses, and SGK3 is shuttered between the nucleus and cytoplasm during the process of fertilized egg cell division, and the altered localization of SGK3 may drive the G2/M transition and mitotic progression in one-cell stage fertilized eggs of the mice. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of hydroxyurea (HU) combined with irradiation on the cycle cycle and apoptosis of the A549 cells after silencing α -thalassemia/mental retardation syndrome X stain-associated protein (ATRX), and to clarify the molecular mechanism. Methods: The A549 cell model (shATRX-A549) stable silencing ATRX was constructed, the infection situation of the cells was observed under fluorescence microscope, the expression of ATRX protein in the ATRX cells was detected by Western blotting method to verify the cell model, and the shNC-A549 cells were chosen as negative control. The experiment were divided into control group, HU group, radiation group (given 8 Gy X-ray radiation), and hydroxyurea+radiation group (given HU+8 Gy X-ray radiation). The percentages of cells at different cell cycle and apoptotic rates of the cells were detected by flow cytometry, and the expressions of mRNA in the cells after silencing ATRX were detected by RNA-sequencing (RNA-seq), and the expression amonts of the cell division cyclin 25B (CDC25B), Cyclin B1, and cyclin dependent kinase 1 (CDK1) proteins in the cells in various groups were detected by Western blotting method. Results: The shNC-A549 cells and shATRX-A549 cells expressed green fluorescence protein (GFP) under fluorescence microscope; the Western blotting results showed that compared with shNC-A549 cells, the expression amount of ATRX protein in the shATRX-A549 cells was decreased. The flow cytometry results showed that compared with control group, the percentage of the shNC-A549 cells at G0/G1 phase in HU group was increased (P<0. 05), the percentages of the shNC-A549 cells at S phase and G2/M phase were significantly decreased (P<0. 05 or P<0. 01), and the percentages of the shNC-A549 cells at G0/G1 phase and S phase in radiation group were significantly decreased (P<0. 01), while the percentage of the shNC-A549 cells at G2/M phage was significantly increased (P<0. 01), the percentage of the shNC-A549 cells at G0/G1 phase in HU+radiation group was significantly decreased (P<0. 01), and the percentages of the cells at S phase and G2/M phase were significantly increased (P<0. 01). Compared with control group, the percentage of the shATRX-A549 cells at G0/G1 phase in HU group was increased (P<0. 05), and the percentage of the shATRX-A549 cells at G2/M phase was decreased (P<0. 01); the percentage of the shATRX-A549 cells at G2/M phase in radiation group was significantly increased (P<0. 01), but the percentages of the shNC-A549 cells at G0/G1 phase and S phase were significantly decreased (P<0. 01), while the percentage of the shATRX-A549 cells at G0/G1 phase in HU+radiation group was decreased (P<0. 01), and the percentages of the shNC-A549 cells at S phase and G2/M phase were significantly increased (P<0. 05). Compared with the shNC-A549 cells, the percentage of the shATRX-A549 cells at G0/G1 phase in radiation group was increased (P<0. 05), the percentage of the shATRX-A549 cells at S phase in HU+radiation group was increased (P<0. 05). Compared with control group, the apoptotic rates of the shNC-A549 cells and shATRX-A549 cells in HU group, radiation group, and HU+radiation group were significantly increased (P<0. 05 or P<0. 01). Compared with shNC-A549 cells, the apoptotic rates of the shATRX-A549 cells in HU group and HU+radiation group were increased （P<0. 05). The differential expressions of mRNAs after silencing ATRX involved c-Myc, Esp1, Cdc20, Plk1, CycA/B, Cip1, and PCNA. The Western blotting results showed that compared with control group, the expression amounts of CDC25B, Cyclin B1, and CDK1 proteins in the shNC-A549 cells and shATRX-A549 cells in HU group, radation group and HU+radiation group were decreased; compared with the shNC-A549 cells, the expressions amounts of Cyclin B1 protein in the shATRX-A549 cells in control and HU groups were decreased, while the expression amounts of CDC25B, cyclin B1, and CDK1 proteins in the shATRX-A549 cells in radiation group and HU+radiation group were increased. Conclusion: HU and radiation can cause the cell cycle arrest and the apoptosis of the A549 cells after silencing ATRX, and its mechanism is related to the CDC25B/Cyclin B/CDK1 pathway. INSET: G0 G1 G2.. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Modern Laboratory Medicine is the property of Journal of Modern Laboratory Medicine Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Spindle assembly checkpoint (SAC) is a protective mechanism for cells to undergo accurate mitosis. SAC prevented chromosome segregation when kinetochores were not, or incorrectly attached to microtubules in the anaphase of mitosis, thus avoiding aneuploid chromosomes in daughter cells. Aneuploidy and altered expression of SAC component proteins are common in different cancers, including lung cancer. Therefore, SAC is a potential new target for lung cancer therapy. Five small molecule inhibitors of monopolar spindle 1 (MPS1), an upstream component protein of SAC, have entered clinical trials. This article introduces the biological functions of SAC, summarizes the abnormal expression of SAC component proteins in various cancers and the research progress of MPS1 inhibitors, and expects to provide a reference for the future development of lung cancer therapeutic strategies targeting SAC components. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To explore the expression and mechanism of cell division cycle associated protein 8 (CDCA8) in the occurrence of prostate cancer (PCa). Methods The difference of CDCA8 mRNA expression levels between normal prostate tissue and PCa tissue was analyzed by bioinformatics method. The disease-free survival (DFS) associated with CDCA8 expression in PCa patients was analyzed using RNA sequencing data from cancer Genome Atlas (TCGA) database. Immunohistochemical method was used to detect the expression of CDCA8 in 56 pairs of PCa tissues and adjacent tissues after radical prostatectomy. Patients were divided into the CDCA8 high expression group (n=31) and the CDCA8 low expression group (n=25) according to staining results. The potential correlation between clinical characteristics and CDCA8 expression level was further analyzed. The CDCA8 gene was silenced by siRNA, and the expression level of CDCA8 mRNA was detected by qPCR. The proliferation of PCa cells was detected by clonal formation assay, and the changes of CDCA8, Ki67, proliferating cell nuclear antigen (PCNA) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway proteins were detected by Western blot assay. Results In TCGA database, CDCA8 mRNA level was significantly higher in PCa tissues than that in normal tissues, and patients with high expression of CDCA8 mRNA had a worse prognosis (P＜0.01). The staining index of expression level of CDCA8 protein was significantly higher in PCa tissue than that in paracancer tissue, and the expression level was positively correlated with total prostate-specific antigen (tPSA), Gleason score, T stage and surgical margin (P＜0.01). The proportion of patients with tPSA≥10 μg/L, T3 stage and positive postoperative resection margin was significantly higher in the high CDCA8 expression group than that in the low CDCA8 expression group (P＜0.05). After CDCA8 gene was silenced, the proliferation of PCa cells was decreased, and the protein expression levels of Ki67, PCNA, p-PI3K and p-AKT were down-regulated (P＜0.01). Conclusion CDCA8 promotes the occurrence of prostate cancer by regulating proliferation, and its mechanism may be related to PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To investigate the function and influence of long non-coding RAN n336928 (LncRNA n336928, n336928) on Gastro-Entero-pancreatic neuroendocrine tumors (GEP-NETs) from the basic-experimental level. Methods: The expression of n336928 in human normal pancreatic ductal epithelial cell lines (hTERT-HPNE) and GEP-Nets cell lines (QGP-1, STC-1) was detected by real-time quantitative fluorescence polymerase chain reaction (qRT-PCR). LncRNA n336928 was knocked down by small interference using transfection reagent Lipo3000 as a carrier. Cell proliferation activity was observed by CCK8 assay, colony assay and EdU assay, cell migration ability was observed by transwell assay, flow cytometry assay was used to detect whether lessen expression of n336928 was related to cell cycle arrest and cell apoptosis, and the expression of related proteins were detected by western blotting. Results: The mRNA expression of n336928 in QGP-1 and STC-1 was significantly higher than that of hTERT-HPNE (P<0.05). After inhibiting the expression of n336928 in QGP-1 and STC-1, the proliferation and migration of the cells were significantly decreased (P<0.05). After transfection of QGP-1 cells with si-n336928, the cell cycle was mainly arrested at G1/S phase, and cell apoptosis was significantly increased. Compared with the control group, cycle-related protein cyclinD1 was significantly decreased in knockout group, while apoptosis-related protein caspase3 was significantly increased (P<0.05). To furtherly investigate the relevant mechanism, RNA immunoprecipitation reaction (RIP) was used to verify that n336928 could bind to EZH2 and jointly regulate the downstream gene P21. Conclusion: LncRNA n336928 can be involved in the development of GEP-NETs by binding to EZH2 and epigeneticly regulation of downstream gene P21. [ABSTRACT FROM AUTHOR]

Objective: To investigate whether sodium butyrate (NaB) and sorafenib synergistically induces ferroptosis to suppress proliferation of hepatocellular carcinoma cells and the possible underlying mechanisms., Methods: CCK8 assay and colony formation assay were used to assess the effects of NaB and sorafenib, alone or in combination, on proliferation of HepG2 cells, and ferroptosis of the treated cells was detected with GSH assay and C11-BODIPY 581/591 fluorescent probe. TCGA database was used to analyze differential YAP gene expression between liver cancer and normal tissues. The effects of NaB and sorafenib on YAP and p-YAP expressions in HepG2 cells were invesitigated using Western blotting., Results: NaB (2 mmol/L) significantly reduced the IC 50 of sorafenib in HepG2 cells, and combination index analysis confirmed the synergy between sorafenib and NaB. The ferroptosis inhibitor Fer-1 and the YAP activator (XMU) obviously reversed the growthinhibitory effects of the combined treatment with NaB and sorafenib in HepG2 cells. The combined treatment with NaB and sorafenib, as compared with the two agents used alone, significantly inhibited colony formation of HepG2 cells, further enhanced cellular shrinkage and dispersion, and decreased intracellular GSH and lipid ROS levels, and these effects were reversed by Fer-1 and XMU. TCGA analysis revealed a higher YAP mRNA expression in liver cancer tissues than in normal liver tissues. NaB combined with sorafenib produced significantly stronger effects than the individual agents for downregulating YAP protein expression and upregulating YAP phosphorylation level in HepG2 cells., Conclusion: NaB combined with sorafenib synergistically inhibit hepatocellular carcinoma cell proliferation possibly by inducing ferroptosis via inhibiting YAP expression.

Objectives: This work aimed to study the correlation between FOXN3-SIN3A complex expression and non-syndromic oral clefts (NSOC) in Xinjiang., Methods: In this study, 60 patients with NSOC attending the People's Hospital of Xinjiang Uygur Autonomous Region were recruited into the case group, including 30 cleft lip with or without cleft palate (NSCL/P), 30 cleft palate only (CPO), and 30 healthy children in the control group. The expression levels of FOXN3, SIN3A, and NEAT1 in peripheral blood of each group were detected by high-throughput second-generation sequencing technology and quantitative reverse transcription polymerase chain reaction (RT-qPCR). Receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to analyze the diagnostic efficiency of NSOC., Results: The comparison of the NSOC and control groups showed that FOXN3, SIN3A, and NEAT1 genes increased compared with the control group. The differences were all statistically significant ( P <0.05). The AUCs of FOXN3, SIN3A, and NEAT1 in the NSCL/P group were 0.933 [95%CI=(0.864, 1.000)], 0.822 [(95%CI=(0.713, 0.932)], and 1.000[95%CI= (1.000, 1.000)], respectively. The AUCs of FOX-N3, SIN3A, and NEAT1 in the CPO group were 0.891 [95%CI=(0.806, 0.976)], 0.688 [95%CI=(0.552, 0.824)], and 1.000 [95%CI=(1.000, 1.000)], respectively., Conclusions: The results showed a correlation between the rising gene expression of FOXN3, SIN3A, and NEAT1 in peripheral blood and the occurrence of NSOC in Xinjiang. This work provides a theoretical basis for further study of the FOXN3-SIN3A complex as biomarkers to facilitate the early screening, disease prediction, and early prevention of NSOC.

To investigate the effect of Inhibitors of protein methyltransferase G9a BIX-01294 on the cell cycle, apoptosis and transplanted tumor. SMMC-7721, BEL-7402, HL-7702 cell lines were subcultured and divided into blank control group and BIX-01294 treatment group with different concentrations (1 滋M, 5 滋M 10 滋M 20 滋M). Western blot was used to detect the expression level of G9a and apoptosis protein CC3, C-PARP, Bax, Bcl-2. The proliferation of SMMC-7721 and BEL-7402 cells treated with BIX-01294 at different concentrations for 24, 48, 72 and 96 hours was detected by Tetramethylazozolium salt (MTT) colorimetry; the cell cycle distribution of hepatoma cells treated with different concentrations of BIX-01294 for 96 hours were examined by flow cytometry; the tumor volume, weight, the protein level of H3K9me2 were measured 21 days after tumor transplantation experiment in nude mice. The expression level of G9a in SMMC-7721 and BEL-7402 cell were higher than HL-7702 cell(P<0.05). Various concentrations of BIX-01294 could inhibit SMMC-7721 cell and BEL-7402 cell proliferation in a time-dependent and dose-dependent manner (P<0.05). There is a significant difference in cell cycle distribution with increased G0 /G1 phage cell percentage, reduced S and G2 /M phage cell percentage at 96h by BIX-01294 treated (P<0.05). The expression levels of CC3, Bax, C-PARP were significantly increased, and the Bcl-2 level were significantly reduced at 96 h by 5 滋M BIX-01294 treated (P<0.05). Compared with the blank control group, the tumor volume and weight of the bix-01294 treated group decreased, and the expression level of H3K9me2 in tumor tissue decreased(P<0.05). BIX-01294 leads to cell cycle arrest and apoptosis of SMMC-7721 and BEL-7402 cells, and has a significant inhibitory effect on tumor growth. It may be through inhibiting the expression of G9a to reduce the expression of H3K9me2 to inhibit tumor growth [ABSTRACT FROM AUTHOR]

Background and objective Cullin1 is a representative member of the Cullin family, and it plays an important role in the ubiquitination of cell cycle, transcription and signal transduction related proteins. Cullin1 is closely related to the occurrence and development of a variety of malignant tumors. The aim of this study is to investigate the effects of Cullin1 on biological function of lung adenocarcinoma A549 and H1395 Cells. Methods The expression of Cullin1 mRNA was detected by quantitative Real-time polymerase chain reaction in lung adenocarcinoma cells (A549, H358, H1395, H1650) and human normal lung epithelial cells BEAS-2B, siRNA technology was used to interfere with lung adenocarcinoma cells with relatively high expression of Cullin1 mRNA; cell proliferation, cell cycle distribution, early cell apoptosis, invasion and migration ability were detected by methyl thiazolyl tetrazolium assay (MTT), flow cytometry and Transwell experiment; Western blot was used to detect the expression levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), Cyclin D1, Cyclin E2, p21 and p27. Results Compared with the BEAS-2B cell, Cullin1 mRNA was highly expressed in lung adenocarcinoma cells, especially in lung adenocarcinoma A549 and H1395 cells (P<0.05). The proliferation ability of lung adenocarcinoma cells was inhibited after interference with Cullin1, and the number of cells in G1 phase increased, the number of cells in S phase decreased, and the early apoptosis rate of lung adenocarcinoma cells is significantly increased (P<0.05); The invasion and migration ability of lung adenocarcinoma cells decreased (P<0.05). After interference with Cullin1, the protein expression of MMP-9, MMP-2, CyclinD1 and CyclinE2 decreased (P<0.05), while the expression of TIMP-1, p21 and p27 protein increased (P<0.05). Conclusion Interference with Cullin1 inhibits the proliferation, invasion and migration of lung adenocarcinoma A549 and H1395 cells, Cullin1 plays a role in promoting cancer in lung adenocarcinoma. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to clarify the relationship between arachidonic acid metabolism, PTEN and its downstream pathways in COX-2 overexpressing colorectal cancer. Methods: By using an over-expressed COX-2 of colorectal cancer cell line CT26 and ApcMin/+mouse adenoma model, combined with arachidonic acid, COX-2 inhibitor NS-398 treatment and COX-2 gene knockout intervention, flow cytometry was used to detect apoptosis, cell cycle and the generation of reactive oxygen species; trans well and colony formation assay were used to test the migration and proliferation capacity of cells; western blot and immunohistochemistry were used to detect the expression of PTEN and its downstream related proteins. Results: In CT26 colorectal cancer, arachidonic acid is metabolized by COX-2 enzymes to produce reactive oxygen species and inactivate PTEN, thereby activating PI3K-AKT protein to promote cell migration and proliferation, and inhibit apoptosis; while COX-2 inhibitor NS-398 prevents the malignant biological behavior of arachidonic acid in CT26 colorectal cancer cells. Moreover, in the COX-2 gene knockout ApcMin/+ mouse adenomas tissue, NS-398 treatment reduced the level of oxidative stress, increased the expression of PTEN, inhibited the phosphorylation of PI3K-AKT, further inhibited the growth of adenomas, and improved the survival rate of mice. Conclusion: Arachidonic acid could produce active oxygen to down-regulate PTEN activity and activate PI3K-AKT to promote colorectal cancer growth through COX-2 enzyme; COX-2 inhibitors can indirectly promote PTEN expression and inhibit colorectal cancer growth, and can act as a potential target for CRC treatment. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of miR-10a inhibition of Tiam1 expression on apoptosis and invasion of gastric cancer cells. Methods: Obtain gastric epithelial tissue cells and gastric cancer tissue cells. QPCR and Western blot experiments were used to detect miR-10a expression and Tiam1 mRNA and protein expression levels in both cells. As well as, the expression of miR-10a and the expression of Tiam1 protein in gastric cancer cell S746T and normal gastric mucosal cells RGM-1 and NGEC were detected. By transfecting miR-10a mimic and miR-10a inhibitor into HS746T cells, flow cytometry was used to detect the cell cycle and apoptosis of HS746T, TranswellTM assay was used to detect the invasion ability of HS746T cells, and qPCR and Western blot were used to detect the apoptosis-related protein caspase3, Caspase9, Bax, and cycle-associated protein P21 expression levels; luciferase activity analysis experiments detected Tiam1 as the target of miR-10a. The constructed Tiam1-highly expressed Tiam1-pcDNA3.1 plasmid and the PX458 plasmid knocked out of the Tiam1 gene were transfected into HS746T cells, respectively, and the apoptosis and invasion ability of HS746T cells were detected by flow cytometry and TranswellTM experiments. Results: Compared with gastric epithelial tissue cells, miR-10a expression was reduced in early gastric cancer clinical tissue cells, and expression of Tiam1 mRNA and protein was increased; The expression of miR-10a is closely related to tumor metastasis in patients with early gastric cancer, and has nothing to do with age, gender and tumor stage; Compared with normal gastric mucosal cells RGM-1 and NGEC, miR-10a expression was reduced in gastric cancer cell HS746T, while Tiam1 protein expression was increased; miR-10a can inhibit the invasion of HS746T cells, promote apoptosis, and stagnate them in G0 / G1 phase; miR-10a targets the 3 'untranslated region (3'UTR) of the Tiam1 gene and reduces Tiam1 protein expression; Tiam1 can inhibit the apoptosis of HS746T cells and promote the invasion of HS746T cells. Conclusion: miR-10a inhibits the proliferation and invasion of HS746T cells and promotes the apoptosis of HS746T cells by targeting the 3' UTR of Tiam1 gene. [ABSTRACT FROM AUTHOR]

Background: Radiation therapy is one of the most common treatments for non-small cell lung cancer (NSCLC). However, the insensitivity of some tumor cells to radiation is one of the major reasons for the poor efficacy of radiotherapy and the poor prognosis of patients, and exploring the underlying mechanisms behind radioresistance is the key to solving this clinical challenge. This study aimed to identify the molecules associated with radioresistance in lung adenocarcinoma (LUAD), identified thyroid hormone receptor interactor 13 (TRIP13) as the main target initially, and explored whether TRIP13 is related to radioresistance in LUAD and the specific mechanism, with the aim of providing theoretical basis and potential targets for the combination therapy of LUAD patients receiving radiotherapy in the clinic., Methods: Three datasets, GSE18842, GSE19188 and GSE33532, were selected from the Gene Expression Omnibus (GEO) database and screened for differentially expressed genes (|log FC|>1.5, P<0.05) in each of the three datasets using the R 4.1.3 software, and then Venn diagram was used to find out the differentially expressed genes common to the three datasets. The screened differential genes were then subjected to protein-protein interaction (PPI) analysis and module analysis with the help of STRING online tool and Cytoscape software, and survival prognosis analysis was performed for each gene with the help of Kaplan-Meier Plotter database, and the TRIP13 gene was identified as the main molecule for subsequent studies. Subsequently, the human LUAD cell line H292 was irradiated with multiple X-rays using a sub-lethal dose irradiation method to construct a radioresistant cell line, H292DR. The radioresistance of H292DR cells was verified using cell counting kit-8 (CCK-8) assay and clone formation assay. The expression levels of TRIP13 in H292 and H292DR cells were measured by Western blot. Small interfering RNA (siRNA) was used to silence the expression of TRIP13 in H292DR cells and Western blot assay was performed. The clone formation ability and migration ability of H292DR cells were observed after TRIP13 silencing, followed by the detection of changes in the expression levels of proteins closely related to homologous recombination, such as ataxia telangiectasia mutated (ATM) protein., Results: Screening of multiple GEO datasets, validation of external datasets and survival analysis revealed that TRIP13 was highly expressed in LUAD and was associated with poor prognosis in LUAD patients who had received radiation therapy. And the results of gene set enrichment analysis (GSEA) of TRIP13 suggested that TRIP13 might be closely associated with LUAD radioresistance by promoting homologous recombination repair after radiation therapy. Experimentally, TRIP13 expression was found to be upregulated in H292DR, and silencing of TRIP13 was able to increase the sensitivity of H292DR cells to radiation., Conclusions: TRIP13 is associated with poor prognosis in LUAD patients treated with radiation, possibly by promoting a homologous recombination repair pathway to mediate resistance of LUAD cells to radiation.

Objective: To explore the role of epigenetic signal molecule bromodomain protein 4 (BRD4) in mediating the therapeutic effect of Liuwei Dihuang (LWDH) Pills on postmenopausal osteoporosis (PMOP) and fatigue., Methods: Thirty rat models of PMOP induced by bilateral ovariectomy were randomized equally into two groups for treatment with normal saline (model group) or LWDH Pills (385.7 mg/kg), with another 15 sham-operated rats as the sham operation group. After 12 weeks of treatment, femoral samples were taken to determine the bone density and BRD4 protein expression. The weight-bearing exhaustive swimming time of the rat models was recorded, and serum levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were measured using enzyme-linked immunosorbent assay. In cultured primary osteoblasts the changes in the expressions of BRD4, MAPK and NF-κB proteins were detected by immunofluorescence staining following treatment with LWDH Pills., Results: The rat models of PMOP showed significantly up-regulated expression of BRD4 protein in the femoral tissue ( P < 0.05), which was obviously lowered by treatment with LWDH Pills. The rats treated with LWDH Pills also showed significant improvement of fatigue. Immunofluorescent staining of the osteoblasts showed that treatment with LWDH Pills significantly decreased the protein expressions of BRD4, MAPK and NF-κB. Analysis of the GSE56116 dataset revealed that that patients with kidney- yin deficiency had significantly higher BRD4 expression than those in the kidney- yang -deficiency group and non-kidney-deficiency group ( P < 0.05). The upregulation of BRD4 expression involved multiple signaling pathways including neural ligand receptor response, cytoskeleton rearrangement, cytokine interaction, and granulocyte colony-stimulating factor chemotaxis pathways., Conclusion: LWDH can alleviate PMOP and fatigue by decreasing BRD4 signaling pathway, suggesting that potential of BRD4 as a promising therapeutic target for PMOP.

Objective: To investigate the expression of WTAP, a m 6 A methylase, in a mouse model of renal ischemia-reperfusion (I/R) injury and the effect of WTAP knockdown on biological behavior of renal tubular epithelial cells exposed to I/R injury., Methods: Sixteen C57BL/6 mice with renal I/R injury or sham operation ( n =8) were examined for blood urea nitrogen (BUN) and creatinine (Scr) levels to assess renal function, and renal pathologies were observed with HE staining. The expressions of WTAP and FOXO1 proteins in the kidneys of the mice were detected using immunohistochemistry. Human renal tubular epithelial cells (HK-2) were transfected with si-WTAP or si-NC followed by hypoxia-reoxygenation (H/R) exposure, Protein and mRNA expression were assessed by Western blot and qRT-PCR, and changes and changes in cell viability and apoptosis were assessed using CCK8 assay and TUNEL staining, respectively; LDH release level and caspase-3 activity of the cells were measured using commercial assay kits. FOXO1 m 6 A modification sites were predicted using SRAMP website (http://www.cuilab.cn/sramp/), and the interaction between WTAP and FOXO1 mRNA was analyzed with RIP experiment; the level of FOXO1 modified by m 6 A was detected by MeRIP-qPCR., Results: Compared with sham-operated mice, the mice with renal I/R injury showed significantly increased Scr and BUN levels ( P < 0.001) and renal expressions of WTAP mRNA and protein ( P < 0.001). In cultured HK-2 cells, H/R exposure significantly decreased the cell viability ( P < 0.001) and increased cellular LDH release ( P < 0.001) and expressions of WTAP mRNA and protein ( P < 0.001). WTAP knockdown obviously reduced the cell damage induced by I/R injury and significantly decreased the mRNA and protein levels of FOXO1 in the cells ( P < 0.001). RIP experiment confirmed WTAP binding to FOXO1 mRNA, and inhibition of WTAP expression significantly reduced FOXO1 m 6 A level in HK-2 cells ( P < 0.001)., Conclusion: WTAP expression is up-regulated in the kidneys of mice with renal I/R injury and in HK-2 cells with H/R exposure. Inhibition of WTAP alleviates H/R-induced apoptotic damage in HK-2 cells possibly by inhibiting FOXO1 expression.

Objective: To investigate the effect of olaparib on melanoma cells and its mechanism. Methods: CCK8 was used to detect the viability of melanoma cells after being treated with olaparib at different concentrations. Western blot was used to detect expression of proteins related with apoptosis and cell cycle in melanoma cells treated with olaparib. Results: Compared with the control group, 5 滋M olaparib can inhibit the viability of melanoma cells (85.53± 2.593)%. And the inhibitory effect of olaparib on the viability of melanoma cells was stronger as its concentration doubling. The viability of melanoma cells were changed to (68.88 ± 1.484)%, (47.21± 1.759)%, (33.04± 1.261)%, (28.17± 1.731)% after the treatment of olaparib at 10 滋M, 20 滋M, 40 滋M, 80 滋M concentrations respectively. Olaparib treatment can promote the expression of apoptosis-related cleaved-PARP1 and inhibit the expression of cell cycle-related cyclin D1 in melanoma cells. Conclusion: Olaparib play a role in inhibiting the viability of melanoma cells by promoting the apoptosis and inhibiting the expression of cell cycle protein. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect and potential mechanism of silencing TCAB1 on the proliferation of human aortic smooth muscle cells(HASMC). Methods: Three si RNA sequences(siTCAB1-331, si TCAB1-619, si TCAB-749) targetingTCAB1 and one negative sequence(NC) were designed, synthesized and then transfected into HASMC with Lipofectamine-TM2000. HASMC were divided three groups: interference group, blank control group, negative control group. Cell transfection was observed under fluorescence microscope. The m RNA and protein expression levels of TCAB1 were detected by RT-q PCR and Western blotting, the best target was selected from the three interfering targets. The effective si RNA(si TCAB1-749) was further transfected into HASMC with LipofectamineTM2000. The changes of cell cycle and CyclinD1 expression of HASMC cells were assessed using flow cytometry,RT-qPCR and Western blotting, respectively. The proliferation of HASMC was assessed using MTS assay after transfecting si TCAB1-749 at 24 h, 48 h,72 h. Results: RT-qPCR and Western blot indicated that the si TCAB1-749 was the best target in the three interfering targets. MTS revealed that the number of cell growth in si TCAB1-749 group was significantly lower than BC group and NCgroup at 24 h, 48 h, 72 h(P<0.05); the percentages of S phases in the cell cycle of si TCAB1-749 group was decreased and G1 phases was increased at 48 h(P < 0.05). In addition, the expression of cyclinD1 was decreased in si TCAB1-749 group(P<0.05). Conclusion:Silencing TCAB1 can inhibit the proliferation of HAVSMC, and its mechanism may be related to the inhibition of Cell cycle protein D1. [ABSTRACT FROM AUTHOR]

Objectives: To explore the possible mechanism of Shao 's five-needle therapy pretreatment on relieving airway inflammatory response in asthmatic rats., Methods: Forty SPF-grade SD rats were randomly divided into a blank group, a model group, an acupuncture group, and a medication group, with 10 rats in each group. Except the blank group, asthma model was established by aerosol inhalation of ovalbumin in the other 3 groups. The rats in the acupuncture group were treated with acupuncture at "Dazhui" (GV 14) and bilateral "Feishu" (BL 13) and "Fengmen" (BL 12), with each session lasting for 20 min. Acupuncture was given before each motivating, once daily for 7 consecutive days. The rats in the medication group were treated with intraperitoneal injection of dexamethasone sodium phosphate solution before each motivating, once daily for 7 days. General situation of the rats was observed in each group; ELISA method was used to detect the levels of inflammatory cytokines interleukin (IL)-1β and IL-18 in serum; immunofluorescence staining method was performed to assess the expression of reactive oxygen species (ROS) in lung tissues; Western blot method was used to measure the protein expression of thioredoxin interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and Caspase-1 in lung tissues., Results: The rats in the blank group exhibited normal behavior, while those in the model group showed signs of respiratory distress, ear scratching, cheek rubbing, and dysphoria. Compared with the model group, the rats in the acupuncture group and the medication group showed stable respiration and relatively agile responses. Compared with those in the blank group, the serum levels of IL-18 and IL-1β were elevated ( P <0.01), the expression intensity of ROS was increased, and the protein expressions of TXNIP, NLRP3, ASC and Caspase-1 in lung tissues were increased ( P <0.01) in the model group. Compared with those in the model group, the serum levels of IL-18 and IL-1β were reduced ( P <0.01), the expression intensity of ROS was lowered, and the protein expressions of TXNIP, NLRP3, ASC and Caspase-1 in lung tissues were reduced (P <0.01) in the acupuncture group and the medication group. Compared with the medication group, the protein expression of ASC in lung tissue was reduced in the acupuncture group ( P <0.05)., Conclusions: Pretreatment of Shao 's five-needle therapy could alleviate airway inflammatory response in asthmatic rats by reducing ROS levels and decreasing the aggregation and activation of pathway-related proteins in the ROS/TXNIP/NLRP3 pathway, ultimately leading to decreased secretion of IL-1β and IL-18. This mechanism may contribute to the effectiveness of Shao 's five-needle therapy in preventing and treating asthma.

Objective: To investigate the effects of methionine restriction on proliferation, cell cycle and apoptosis of human acute leukemia cells., Methods: Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of methionine restriction on HL-60 and Jurkat cells proliferation. The effect of methionine restriction on cell cycle of HL-60 and Jurkat cells was examined by PI staining. Annexin V-FITC / PI double staining was applied to detect apoptosis of HL-60 and Jurkat cells following methionine restriction. The expression of cell cycle-related proteins cyclin B1, CDC2 and apoptosis-related protein Bcl-2 was evaluated by Western blot assay., Results: Methionine restriction significantly inhibited the proliferation of HL-60 and Jurkat cells in a time-dependent manner (HL-60: r =0.7773, Jurkat: r =0.8725), arrested the cells at G2/M phase ( P < 0.001), and significantly induced apoptosis of HL-60 and Jurkat cells (HL-60: P < 0.001; Jurkat: P < 0.05). Furthermore, Western blot analysis demonstrated that methionine restriction significantly reduced the proteins expression of Cyclin B1 ( P < 0.05), CDC2 ( P < 0.01) and Bcl-2 ( P < 0.001) in HL-60 and Jurkat cells., Conclusion: Acute leukemia cells HL-60 and Jurkat exhibit methionine dependence. Methionine restriction can significantly inhibit the proliferation, promote cell cycle arrest and induce apoptosis of HL-60 and Jurkat cells, which suggests that methionine restriction may be a potential therapeutic strategy for acute leukemia.

Recent studies have shown that the occurrence and development of common liver diseases, including non-alcoholic fatty liver disease, cirrhosis, and liver cancer, are related to liver aging(LA). Therefore, to explore the effect and mechanism of Dahuang Zhechong Pills(DHZCP), a traditional classic prescription in improving LA with multiple targets, the present study randomly divided 24 rats into a normal group, a model group, a DHZCP group, and a vitamin E(VE) group, with six rats in each group. The LA model was induced by continuous intraperitoneal injection of D-galactose(D-gal) in rats. For the LA model rats, the general situation was evaluated by aging phenotype and body weight(BW). LA was assessed by the pathological characteristics of hepatocyte senescence, hepatic function indexes, the staining characteristics of phosphorylated histone family 2A variant(γ-H2AX), and the expression levels of cell cycle arrest proteins(P21, P53, P16) and senescence-associated secretory phenotype(SASP) in the liver. The activation of the reactive oxygen species(ROS)-mediated phosphatidylinositol-3 kinase(PI3K)/protein kinase B(Akt)/forkhead box protein O4(FoxO4) signaling pathway was estimated by hepatic ROS expression feature and the protein expression levels of the key signaling molecules in the PI3K/Akt/FoxO4 signaling pathway. The results showed that after the treatment with DHZCP or VE for 12 weeks, for the DHZCP and VE groups, the characterized aging phenotype, BW, pathological characteristics of hepatocyte senescence, hepatic function indexes, relative expression of ROS in the liver, protein expression levels of key signaling molecules including p-PI3K, p-Akt, and FoxO4 in the liver, staining characteristics of γ-H2AX, and the protein expression levels of P16, P21, P53, interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in the liver were improved, and the effects of DHZCP and VE were similar. Based on the D-gal-induced LA model in rats, this study demonstrates that DHZCP can ameliorate LA with multiple targets in vivo, and its effects and mechanism are related to regulating the activation of the ROS-mediated PI3K/Akt/FoxO4 signaling pathway in the liver. These findings are expected to provide new pharmacological evidence for the treatment of DHZCP in aging-related liver diseases.

Objective: To investigate the effects of mTOR inhibitors everolimus (EVE) and gemcitabine (GEM) on the proliferation, apoptosis and cell cycle of diffuse large B-cell lymphoma (DLBCL) cell line U2932, and further explore the molecular mechanisms, so as to provide new ideas and experimental basis for the clinical treatment of DLBCL., Methods: The effect of EVE and GEM on the proliferation of U2932 cells was detected by CCK-8 assay, the IC 50 of the two drugs was calculated, and the combination index ( CI =) of the two drugs was calculated by CompuSyn software. The effect of EVE and GEM on apoptosis of U2932 cells was detected by flow cytometry with AnnexinV-FITC/PI staining. Flow cytometry with propidium iodide (PI) staining was used to detect the effect of EVE and GEM on the cell cycle of U2932 cells. Western blot assay was used to detect the effects of EVE and GEM on the channel proteins p-mTOR and p-4EBP1, the anti-apoptotic proteins MCL-1 and Survivin, and the cell cycle protein Cyclin D1., Results: Both EVE and GEM could significantly inhitbit the proliferation of U2932 cells in a time- and dose-dependent manner ( r =0.465, 0.848; 0.555, 0.796). According to the calculation of CompuSyn software, EVE combined with GEM inhibited the proliferation of U2932 cells at 24, 48 and 72 h with CI =<1, which had a synergistic effect. After treated U2932 cells with 10 nmol/L EVE, 250 nmol/L GEM alone and in combination for 48 h, both EVE and GEM induced apoptosis, and the difference was statistically significant compared with the control group ( P <0.05). The apoptosis rate was significantly enhanced after EVE in combination with GEM compared with single-agent ( P <0.05). Both EVE and GEM alone and in combination significantly increased the proportion of cells in G 1 phase compared with the control group ( P <0.05). The proportion of cells in G 1 phase was significantly increased when the two drugs were combined ( P <0.05). The expression of p-mTOR and effector protein p-4EBP1 was significantly downregulated in the EVE combined with GEM group, the expression of anti-apoptotic proteins MCL-1, Survivin and cell cycle protein cyclin D1 was downregulated too ( P <0.05)., Conclusion: EVE combined with GEM can synergistically inhibit the proliferation of U2932 cells, and the mechanism may be that they can synergistically induce apoptosis by downregulating the expression of MCL-1 and Survivin proteins and block the cell cycle progression by downregulating the expression of Cyclin D1.

Objective: To investigate the in vitro inhibitory activity of a novel class Ⅰ and Ⅱb selective histone deacetylase (HDAC) inhibitor, purinostat mesylate (PM) , in diffuse large B-cell lymphoma and its mechanism. Methods: The 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide method was used to detect the effect of PM on cell proliferation. The effects of PM on cell cycle and apoptosis were detected by flow cytometry. The acetylation levels of HDAC substrate, cell cycle protein, apoptosis-related protein, and oncogene protein expression were detected by Western blot. Results: PM significantly inhibited the proliferation of lymphoma SUDHL-4 and SUDHL-6 cells and increased the acetylation levels of HDAC substrates H3, H4, and α-tubulin. In cell cycle experiments, PM induced G(0)/G(1) phase arrest in SUDHL-4 and SUDHL-6 cells. Western blot experiment showed that PM could significantly downregulate the expression of cyclin-dependent kinases Cdk2, Cdk4, Cdk6, cyclin D1, and cyclin E and upregulate the expression of CDK inhibitor protein p21. In the apoptosis experiment, PM could induce the apoptosis of SUDHL-4 and SUDHL-6 cells. Western blot experiment demonstrated that PM promoted endogenous apoptosis by activating caspase-3 kinase and affecting antiapoptotic protein Bcl-2. In addition, PM could downregulate the expression of oncogene marker proteins MYC, IKZF1, and IKZF3. Conclusion: PM has an efficient biological activity in vitro for diffuse large B-cell lymphoma, including double-hit lymphoma, and provides valuable experimental evidence for PM in clinical treatment.

Objective To investigate how mutation of nuclear autoantigenic sperm protein (NASP) gene affects mouse liver fibrosis induced by concanavalinA (ConA) and its mechanism. Methods The wild-type B6 (B6-WT) mice were used as a control group, and the NASP mutant B6 (B6-NASP M ) mice as an experimental group. The mice were injected with ConA via tail vein once a week for 8 weeks to establish the model of liver fibrosis. The histopathological changes were observed by HE staining, the collagen fiber deposition by Masson staining, smooth muscle alpha-actin (α-SMA) expression in liver tissue by immunohistochemical staining. The levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by microplate assay. The serum tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) were measured by ELISA. The mRNA expressions of type I collagen (Col1) and Col3 in liver tissue were determined by real-time quantitative PCR, and the changes of T lymphocyte subsets in liver tissue were detected by flow cytometry. Results Compared with the B6-WT group, B6-NASP M group had disordered liver structure and no significant changes in mRNA expression of Col 1 and Col3, but the collagen fiber hyperplasia and α-SMA expression in liver tissue were more obvious, and the levels of serum ALT and TNF-α were significantly increased. In addition, the proportion and number of CD4 + CD44 hi T lymphocyte subsets in liver tissue were markedly decreased. Conclusion Mutation of NASP gene aggravates mouse liver fibrosis by increasing the release of TNF-α, changing the proportion of T lymphocyte subsets in liver tissue and promoting the activation of hepatic stellate cells.

Objective: To analyze the clinical characteristics and pathogenic gene in a Chinese pedigree affected with mitochondrial DNA depletion syndrome 8A (MTDPS8A)., Methods: Whole exome sequencing was carried out for the patient. Sanger sequencing was used to verify the results, and PolyPhen-2 and PROVEAN software were used to predict the impact of amino acid changes on the function of the protein., Results: The patient, a two-month-old female, was admitted to the hospital for poor milk intake and poor mental response. Her clinical manifestations included feeding difficulty, shortness of breath and low muscle tone. Auxiliary laboratory test indicated that the infant was underdeveloped with abnormal liver, kidney, and heart functions accompanied by hyperlacticacidemia. She responded poorly to treatment and eventually died. Sequencing revealed that the child has carried compound heterozygous missense variants of the RRM2B gene, namely c.16delA (p.R6Gfs*22) and c.175G>C (p.A59P), which were respectively inherited from her father and mother, and both were newly discovered pathologic variants., Conclusion: The c.16delA and c.175G>C compound heterozygous variants of the RRM2B gene probably underlay the pathogenesis of MTDPS8A. Above finding has strengthened the understanding of the clinical feature and genetic etiology of this disease and expanded the mutation spectrum of the RRM2B gene.

Objective To investigate the effect of lncRNA CRNDE on proliferation, apoptosis, and cell cycle of U937 cells and its mechanism. Methods The expression level of CRNDE in bone marrow cells of AML patients was analyzed by GEPIA database; the mRNA expression levels of miR-136-5p, CRNDE, and minichromosome maintenance 5(MCM5) in AML cell lines were detected by quantitative real-time PCR (qRT-PCR). The lentiviral vector with CRNDE knocked down was constructed and transfected into U937 cells which were randomized into CRNDE knockdown group (sh-CRNDE group) and negative control group (sh-NC group); miR-136-5p mimic and miR-136-5p inhibitor were transfected respectively to overexpress and knock down miR-136-5p in U937 cells which were randomized into miR-136-5p-mimic group, NC-mimic group, miR-136-5p-inhibitor group, and NC-inhibitor group. The effect of CRNDE and miR-136-5p on proliferation was detected by CCK-8 assay and cell counting assay, and the effect of them on cell cycle and apoptosis was detected by flow cytometry. The mRNA expressions of miR-136-5p, CRNDE, and MCM5 were detected by qRT-PCR, and the protein expressions of MCM5, Bcl2, cyclin D1, and cyclin A2 were detected by Western blotting. Results CRNDE was highly expressed in the bone marrow and cell lines of AML patients. Knockdown of CRNDE upregulated miR-136-5p, inhibited the MCM5 mRNA and protein expressions and the cell proliferation, promoted the cell apoptosis, and blocked the cell cycle in G1 phase. Overexpression of miR-136-5p also inhibited the expression of MCM5 at both mRNA and protein levels, while knockdown of miR-136-5p reversed those effects. Conclusion CRNDE promotes the proliferation and inhibits the apoptosis of U937 cells by downregulating miR-136-5p and upregulating MCM5.

Objective: To investigate the relationship between hypoxia inducible factor 1 (HIF1α) and Wilms' tumor 1associating protein (WTAP) expression level in t(8;21) acute myeloid leukemia cells., Methods: The t(8;21) acute myeloid leukemia cell lines, including SKNO-1 and Kasumi-1 were treated by Echinomycin for 24 h, RT-qPCR and Western blot were used to detect the expression levels of WTAP mRNA and the protein. The CoCl 2 was used to induce the hypoxia of the cells for 24 h, the expression levels of HIF1α, WTAP protein were detected by Western blot., Results: The expression level of WTAP mRNA and the protein in the echinomycin treated group was significantly lower than those in the control group (P<0.01). The expression level of WTAP protein in the CoCl 2 treated group was significantly higher than those in the control group (P<0.05)., Conclusion: The inhibition of HIF1-α could down-regulates the expression of WTAP, while the up-regulation of HIF1α could up-regulates the expression of WTAP, which shows that there is a positive correlation of HIF1α and WTAP expression. This result suggesting that HIF1α may be involves in the expression regulation of WTAP gene.

Objective: To explore the regulatory effect of TRIP13 on the proliferation and apoptosis of B-cell lymphoma cells and its possible molecular mechanism by knocking down/overexpressing TRIP13 on the cell lines Granta-519 and JVM-2., Methods: Lentiviral transfection technology was used to construct Granta-519 and JVM-2 cells with knocked down or overexpressed TRIP13 and their control cells. The efficiency of transfection was determined by fluorescence microscopy. The efficiency of knockdown and overexpression was evaluated by real-time quantitative PCR and Western blot. The proliferation was detected by CCK-8 assay. The apoptosis was detected by the Annexin V-APC single staining. The cell cycle was detected by the PI staining. The expression levels of P53, MDM4, and BCL-2 were evaluated by Western blot., Results: After TRIP13 was knocked down, the proliferation ability of Granta-519 and JVM-2 cells was significantly reduced, and the apoptosis rate significantly increased. After TRIP13 was overexpressed, the proliferation ability of Granta-519 and JVM-2 cells was significantly enhanced, and the apoptosis was significantly reduced. After TRIP13 was knocked down, Granta-519 cells had obvious G 1 phase arrest, and JVM-2 cells had obvious G 1 and G 2 /M phase arrest. After TRIP13 was knocked down in Granta-519 cells, the expression of BCL-2 protein decreased, while MDM4 protein increased. After TRIP13 was overexpressed, the expression of MDM4 protein decreased. After TRIP13 was overexpressed in JVM-2 cells, the expression of BCL-2 protein increased., Conclusion: TRIP13 promotes the proliferation of B-cell lymphoma cells, inhibits their apoptosis, and affects their proliferation and apoptosis by participating in the regulation of the cell cycle. TRIP13 promotes the expression of BCL-2 proteins and inhibits the expression of MDM4 protein in B-cell lymphoma cells.

Objective: To investigate the expression of cell division cycle protein 37 (Cdc37) in multiple myeloma (MM) and its effect on MM cell proliferation., Methods: The expression of Cdc37 mRNA in CD138 + cells derived from 63 newly diagnosed MM patients and 8 healthy people were detected by real-time quantitative PCR (RT-qPCR). Cdc37 was down-regulated by lentivirus in MM cell line NCI-H929. CCK-8 assay and soft agar clone formation assay were conducted to explore the role of Cdc37 on MM proliferation in vitro. To further verify the effect of Cdc37 on MM cell proliferation in vivo, NOD/SCID mice subcutaneous tumorigenesis model was established. Flow cytometry was carried out to explore the role of Cdc37 on cell cycle. Cell cycle associated proteins and NF-κB pathway were detected by Western blot (WB)., Results: Cdc37 was highly expressed in newly diagnosed CD138 + cells compared with healthy people. After Cdc37 suppression by shRNA lentivirus infection in NCI-H929 cells, the proliferation of MM cells were decreased in vitro and in vivo. Compared with the control group, the ratio of cells arrested in G 0 /G 1 phase significantly increased in NCI-H929-Cdc37 shRNA cells, the expression of cyclin D1 decreased, while the expression of p21 and p53 was significantly up-regulated. Meanwhile, the activation of NF-κB signaling pathway was hampered in NCI-H929-Cdc37 shRNA cells., Conclusion: Cdc37 is highly expressed in newly diagnosed MM patients. Inhibition of Cdc37 results in decreased proliferation activity and G 0 /G 1 arrest in NCI-H929 cells. The possible mechanism may be to inhibit the activation of NF-κB signaling pathway.

Objective: To explore the role of small nuclear noncoding RNA 7SK in embryonic stem cell (ESCs) proliferation and the value of 7SK as a target for early diagnosis and treatment for primordial dwarfism (PD)., Methods: ESC line R1 was transfected with the CRISPR/Cas9 system, and sequencing of the PCR product and glycerol gradient analysis were performed to identify novel 7SK deletion mutations. A lentivirus system was used to knock down cyclin-dependent kinase 9 (CDK9) in clones with 7SK deletion mutations, and the effect of CDK9 knockdown on the protein level of cell division cycle 6 (CDC6) was analyzed with Western blotting., Results: We identified a novel deletion mutation of 7SK at 128-179 nt in the ESCs, which resulted in deficiency of cell proliferation. 7SK truncation at 128-179 nt significantly reduced the protein expressions of La-related protein 7 (LARP7) and CDC6., Conclusions: 7SK truncation at 128-179 nt can significantly impair proliferation of ESCs by downregulating CDC6. 7SK is a key regulator of proliferation and mediates the growth of ESCs through a mechanism dependent on CDK9 activity, suggesting the value of 7SK truncation at 128-179 nt as a potential target for early diagnosis and treatment of PD.

Bromodomain-containing protein 4 (BRD4) is one of the most important members in the bromodomain and extra terminal domain(BET) family, it plays an important role in cellular physiology in human body, such as cell cycles, cell proliferation, and immune response. Recent studies have shown that BRD4 is associated with occurrence and development of acute myeloblastic leukemia, multiple myeloma and lymphoma. The mechanisms of BRD4 in hematologic malignancies including the regulation of c-Myc expression, and participation of the composition of super-enhancer, etc. At present, many kinds of inhibitors have been developed to target inhibit BRD4 for therapy in hematologic malignancies, and some of BRD4 inhibitors have entered phase Ⅱ clinical trials, which suggested that BRD4 inhibitors are expected to become new therapeutic agents for hematologic malignancies. In this review, the research advance of BRD4 and BRD4 inhibitors in hematologic malignancies was summarized briefly.

FAM60A,a cell cycle protein,is a subunit of the SIN3 transcription regulator family member A/histone deacetylase(SIN3-HDAC)complex and plays an important role in cell cycle regulation,cell morphology change,cell proliferation,differentiation and migration,early embryogenesis and so on.Studies in recent years have shown that FAM60A plays a role in the occurrence and development of tumors including human osteosarcoma,esophageal cancer,gastric cancer,lung cancer and liver cancer,providing a new research direction for tumor diagnosis and treatment.Based on the research results in recent years at home and abroad,this paper discussed the effects of FAM60A on cellular functions.

Objective: To investigate the expression of WTAP gene in acute myeloid leukemia (AML) and its clinical significance., Methods: 74 acute myeloid leukemia patients with non-M3 type and 19 normal donors were selected, and real-time quantitative polymerase chain reaction was used to detect the mRNA expression level of WTAP gene in their bone marrow cells. The relationship between the mRNA expression level of WTAP gene and the clinical characteristics was analyzed., Results: The relative mRNA expression of WTAP gene in the non-M3 AML group was significantly higher than that in the healthy control group, and the difference showed statistically significant (P<0.01). There showed no statistically significant difference in WTAP gene expression among each subtypes (all P>0.05) according to the classification of FAB. The mRNA expression level of WTAP gene in FLT3-ITD mutated AML patients was higher than that in FLT3-ITD unmutated group (P=0.016), and the mRNA expression level of WTAP gene in AML patients with CEBPα mutation was lower than that in CEBPα unmutated group (P=0.016). The expression level of WTAP mRNA was positively correlated with WT1 expression (r=0.6866, P<0.01). There was no relationship between WTAP mRNA expression level and other clinical parameters, such as age, gender, white blood cell count, hemoglobin level, platelet count, bone marrow original proportion of immature cells, chromosome karyotype, and NPM1, DNMT3A, ASXL1, NRAS, TET2 genes mutation status (P>0.05). The expression level of WTAP mRNA showed no obvious effect on the complete remission of patients after first treatment. The different expression level of WTAP gene at initial diagnosis showed also no effect on the overall survival time of patients., Conclusion: The expression level of WTAP gene is increasing in new diagnosed non-M3 acute myeloid leukemia. There is a positive correlation between the expression level of WTAP gene and the expression level of WT1 fusion gene. WTAP mRNA always shows higher expression in patients with FLT3-ITD mutation than that in patients without FLT3-ITD mutation, and shows lower expression in patients with CEBPα mutation than that in unmutated group.

Objective: To investigate the effect of centrosomal protein Cep63 on the apoptosis of papillary thyroid carcinoma (PTC) cell lines TPC-1 and underlying mechanism. Methods: With collected PTC tissues and adjacent tissues, Cep63 expression was detected by RT-qPCR and its relationship with clinicopathological factors was analyzed. The experiment included negative control group (NC), low expression group (Cep63(-)) and overexpression group (Cep63(+)), and wild-type TPC-1 cells were transfected with Cep63 lentivirus. The efficiency of Cep63 was detected by western blot (WB) and qRT-PCR. Cell proliferation ability was detected by plate cloning experiment and MTT assay. Cell apoptotic rate was detected by flow cytometry, and expression levels of apoptosis-related proteins were detected by immunohistochemistry and WB. The t-test was used to compare the differences in the means between the two groups, the one-way analysis of variance was used to compare multiple groups, and the chi-square test was used to analyze the association between gene expression levels and pathological factors. Results: Compared with NC group, cell proliferation ability was significantly decreased in Cep63(-) group (3.18±0.07 vs. 2.14±0.09, t =8.54, P <0.01) and significantly increased in Cep63(+) group (3.18±0.07 vs. 3.58±0.10, t =3.21, P <0.05). Apoptotic rates in NC, Cep63 (-) and Cep63 (+) groups were respectively 3.03%±0.24%, 8.66%±0.44% and 1.17%±0.44%, and the flow cytometry showed that the low expression of Cep63 s ignificantly increased the apoptosis TPC-1 cells ( F =157.7, P <0.001). Bcl-2 protein expression levels of NC, Cep63 (-) and Cep63 (+) groups were respectively 1.07±0.03, 0.49±0.01 and 1.99±0.09, and BAX protein expression levels of three groups were respectively 0.64±0.02, 1.06±0.01 and 0.21±0.03. WB showed that the expression level of Bcl-2 decreased ( F =183.2, P <0.001), while the expression level of BAX was significantly up-regulated ( F =283.7, P <0.001). Conclusion: Cep63 may regulate the apoptotic process of TPC-1 cells through Bcl-2/BAX pathway and Cep63 may be a potential oncogene of PTC.

Objective: To investigate the expressions of HIF-1α, BRD4, Beclin1, LC3B and p62 in breast cancer tissues and their clinicopathological significance, and to study alterations of their expression in breast cancer cells under hypoxic microenvironment. Methods: Immunohistochemistry was used to detect HIF-1α, BRD4, Beclin1, LC3B and p62 protein expressions in 125 breast cancer tissues and 50 para-cancer normal breast tissues, and their correlation with clinicopathologic characteristics were analyzed. The expression of these proteins were also measured after 24 hours of hypoxia stimulation was detected in different breast cancer cell lines and normal breast epithelial cells. Results: The expression of HIF-1α, BRD4, Beclin1 and LC3B proteins in breast cancer tissues were significantly higher than in para-cancer normal breast tissues ( P <0.05). There was a positive association between histologic grade, the expression of HIF-1α, BRD4, Beclin1 and LC3B ( P <0.05). High expressions of HIF-1a and Beclin1 were often correlated with lymph node metastasis and lymphatic invasion ( P <0.05). Increased HIF-1α, BRD4, Beclin1 and LC3B expression was associated with ER or PR negativity, but only HIF-1α was associated with HER2 positivity ( P <0.05). HIF-1α, BRD4, Beclin1, and LC3B were positively correlated with each other in breast cancer tissues ( P <0.01). After 24 hours of hypoxic stimulation, the expression of HIF-1α, BRD4, Beclin1 and LC3B was up-regulated in breast cancer cells. Conclusions: Hypoxia induces autophagy in breast cancer tissues. HIF-1α is positively correlated with BRD4, suggesting that BRD4 is involved in the regulation of autophagy by hypoxic microenvironment in breast cancer. High expression of HIF-1α, BRD4 and autophagy may play an important role in the development of breast cancer.

Objective: The microRNA (miRNA) prognostic model can predict the prognosis of patients with oral squamous cell carcinoma (OSCC) on the basis of bioinformatics. Moreover, it can accurately group OSCC patients to improve targeted treatment., Methods: We downloaded the miRNA and mRNA expression profile and clinical data of OSCC from The Cancer Genome Atlas (TCGA). The risk score model of miRNA was screened and established by univariate and multivariate Cox regression models. The performance of this prognostic model was tested by receiver operating characteristic (ROC) curves and area under the curve (AUC). The target genes of six miRNAs were predicted and intersected with differential mRNA for enrichment analysis by Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway and gene ontology (GO) enrichment analysis. A protein protein interaction network (PPI) was constructed to screen hub genes., Results: By using univariate and multivariate Cox regression analyses, the prognostic risk model was obtained. The AUC of the ROC curve for predicting 5-year survival in the training group, test group, and whole cohort were 0.757, 0.673, and 0.724, respectively. Furthermore, univariate Cox regression and multivariate Cox regression considering other clinical factors showed that the six-miRNAs signature could serve as an independent prognostic factor (P<0.001). The top 10 hub genes in the PPI network screened by intersecting target genes include CCNB1, EGF, KIF23, MCM10, ITGAV, MELK, PLK4, ADCY2, CENPF, and TRIP13. EGF and ADCY2 were associated with survival prognosis (P<0.05)., Conclusions: The six-miRNAs signature could efficiently function as a novel and independent prognostic model for OSCC patients, which may be a new method to guide the accurate targeting treatment of OSCC.

Purpose: To investigate the expression of metalloproteinase-14 (MMP-14), growth inhibitory factor 4 (ING4) and hypoxia-inducible factor-1α (HIF-1α) in human oral squamous cell carcinoma (OSCC) and their clinical significance, to provide references for clinical diagnosis and treatment of OSCC., Methods: Sixty patients with OSCC admitted to our hospital from July 2014 to July 2019 were enrolled in this study. The tissue specimen of 60 patients were prepared into paraffin sections as the experimental group, while 20 cases of normal oral mucosal tissue were paraffin-embedded as the control group. Immunohistochemical S-P method was used to analyze the positive expression levels of MMP-14, ING4 and HIF-1α in oral mucosa of the two groups. The relationship between the 3 indicators and the different clinicopathological features of OSCC patients was analyzed, the correlation of the expression level between 3 protein was also calculated. SPSS 23.0 software package was used for statistical analysis of the data., Results: The positive expression rate of ING4 in OSCC patients was 41.7% (25/60), which was significantly lower than 70.0% (14/20) in the control group (P<0.05). The positive expression rates of MMP-14 and HIF-1α were 68.3% (41/60), 71.7% (43/60), respectively, significantly higher than 20.0% (4/20) and 15.0% (3/20) of the control group. The difference between the 2 groups was statistically significant (P<0.01). The positive expression rate of the 3 proteins was significantly correlated with TNM stage, differentiation and cervical lymph node metastasis (P<0.05). In OSCC tissues, there was a significant negative correlation between ING4 and HIF-1α or MMP-14 expression (r=-0.39, -0.51, P<0.01), while HIF-1α and MMP-14 expression was significantly positively correlated with the clinical features (r= 0.45, P<0.01)., Conclusions: MMP-14, ING4 and HIF-1α are closely related to the progression of OSCC. Down-regulation of ING4 can promote up-regulation of MMP-14 and HIF-1α, and ultimately promote formation, growth, invasion and metastasis of OSCC. The expression levels of the 3 proteins can be used as important indicators for judging the clinical features and prognosis of patients with OSCC.

The purpose of the present study was to investigate the effects of forkhead box O4 (FOXO4) on the senescence of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). The hUC-MSCs were induced to senescence by natural passage, and FOXO4 expression was inhibited by lentiviral shRNA transfection. The hallmark of cell senescence was analyzed by β-galactosidase staining, and the cell viability was assayed by CCK-8 method. Flow cytometry was used to investigate the apoptosis of hUC-MSCs. The expression levels of Bcl-2, Bax, FOXO4, interleukin 6 (IL-6) and cleaved Caspase-3 were detected by qPCR and Western blot. Immunofluorescence staining was used to detect FOXO4 expression. The amount of IL-6 secreted by hUC-MSCs was detected by ELISA. The results showed that, compared with the passage 1, senescent hUC-MSCs showed up-regulated expression levels of Bax and FOXO4, down-regulated expression levels of Bcl-2 and cleaved Caspase-3, and increased IL-6 mRNA expression and secretion. FOXO4 inhibition in senescent hUC-MSCs promoted cell apoptosis, reduced cell viability, and inhibited the mRNA expression and secretion of IL-6. These results suggest that FOXO4 maintains viability and function of senescent hUC-MSCs by repressing their apoptosis response, thus accelerating senescence of the whole cell colony.

Objective: To explore the role of cell division cycle protein 37 (Cdc37) mediating bortezomib (BTZ) resistance in multiple myeloma (MM) via the regulation of autophagy activity to provide a novel strategy for MM therapy. Methods: The expressions of Cdc37 and LC3b were investigated in BTZ-resistant MM cell line ANBL-6.BR using quantitative real-time PCR (qRT-PCR) and western blot (WB) analysis. Cdc37 was upregulated in ANBL-6.BR cells owing to lentivirus transfection. The LC3b expression was detected with WB, and BTZ-induced apoptosis was explored using flow cytometry. Cdc37 was then down-regulated by shRNA in the MM cell line NCI-H929. Sensitivity of BTZ was evaluated using CCK-8 analysis. WB analysis was performed to check the expression of the AKT/mTOR pathway and autophagy-associated proteins. The sensitivity of NCI-H929 cells to BTZ in the presence of autophagy inhibitor chloroquine (CQ) was analyzed using flow cytometry. Results: Cdc37 was down-regulated, while autophagy-associated gene LC3b was upregulated in BTZ-resistant cell line ANBL-6.BR. Up-regulated Cdc37 in ANBL-6.BR cells could inhibit LC3b expression and increase the sensitivity of MM to BTZ. Suppressing Cdc37 expression in MM cell line NCI-H929 induced BTZ resistance and autophagy activation, while CQ could rescue BTZ resistance caused by Cdc37 inhibition. Conclusion: Cdc37 may participate in BTZ resistance in MM via the regulation of autophagy activity.

Objective To analyze the expression of splicing factors in gastric cancer using bioinformatics methods and investigate the effect of aberrantly expressed serine/arginine-rich splicing factor(SRSF10)on the phenotype of gastric cancer cells. Methods The RNA-seq data of gastric cancer and paracancerous tissues were downloaded from The Cancer Genome Atlas(TCGA)cancer database,and bioinformatics analysis was performed to obtain the splicing factors differentially expressed in gastric cancer.The splicing factor SRSF10 was selected to investigate its effect on the development of gastric cancer.RNA interference technology was used to construct SRSF10 knockdown gastric cancer cells.MTS,Transwell,and cell scratches were used to study the effect of SRSF10 knockdown on gastric cancer cell phenotype. Results A total of 48 splicing factors were identified in gastric cancer by a series of bioinformatics techniques,of which 35 were up-regulated and 13 were down-regulated.The splicing factor SRSF10,which was up-regulated,was selected for further study.It was found that the gastric cancer cells after SRSF10 knockdown proliferated more slowly and had lower migration ability than normal gastric cancer cells. Conclusions Multiple splicing factors are found in gastric cancer and may play an important role in the development of gastric cancer.The splicing factor SRSF10 may contribute to the pathogenesis of gastric cancer.

Objective: This study aimed to explore the target relationship between miR-204-5p and bromodomain-containing protein (BRD) 4, as well as their effects on cell proliferation, migration, and invasion in tongue squamous cell carcinoma SCC25., Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to detect miR-204-5p and BRD4 expression levels in tongue squamous cell carcinoma and different cell lines. TargetScan and dual luciferase reporter assay were used to confirm the target relationship between miR-204-5p and BRD4. The effects of miR-204-5p on SCC25 cell proliferation were examined by cell counting kit (CCK) 8 assay, whereas those on SCC25 cell migration and invasion were determined by Transwell assay. RT-qPCR and Western blot were used to detect the effects of miR-204-5p mimics and inhibitors on BRD4 expression. Transwell and CCK8 assays were used to detect the effects of miR-204-5p on proliferation, migration, and invasion through BRD4 regulation., Results: miR-204-5p was significantly downregulated in the tissues and cells of squamous cell carcinoma, and BRD4 showed the opposite result. The increase in miR-204-5p expression can inhibit the proliferation, migration, and invasion of SCC25 cells. TargetScan and luciferase test confirmed that miR-204-5p and BRD4 had a negative regulatory relationship with BRD4, respectively. Moreover, miR-204-5p mimics can inhibit BRD4 expression, and miR-204-5p inhibitors can promote BRD4 expression upregulation. When miR-204-5p and BRD4 were overexpressed in SCC25 cells, BRD4 can make up for the inhibitory effect of miR-204-5p on SCC25 cells., Conclusions: miR-204-5p could inhibit proliferation, migration and invasion in tongue squamous cell carcinoma SCC25 cells by targeting BRD4 gene.

Objective: To examine the expression of vasohibin-1, metastasis-associated in colon cancer-1 (MACC1) and KAI1 proteins in serous ovarian cancer and their clinical significance.
 Methods: In 124 specimens of serous ovarian cancer (serous ovarian cancer group) and 30 specimens of ovarian serous cystadenoma (ovarian serous cystadenoma group), the expression of vasohibin-1, MACC1 and KAI1 protiens were detected by immunohistochemistry ElivisionTM method.
 Results: In the serous ovarian cancer group, the positive rates of vasohibin-1 and MACC1 proteins were 48.4% and 58.1%, respectively, which were both higher than those in the ovarian serous cystadenoma group (10.0% and 13.3%, respectively); while the positive rate of KAI1 protein in the serous ovarian cancer group was 33.9%, which was lower than that in the ovarian serous cystadenoma group (86.7%), there were significant differences between the 2 groups (all P<0.05). In the serous ovarian cancer group, the expression of the 3 proteins were closely related to the pathological grade, Federation International of Gynecology and Obstetrics (FIGO) stage and pelvic lymph node metastasis (all P<0.05). The KAI1 protein was negatively correlated with the levels of vasohibin-1 and MACC1 (r=-0.500, -0.600, respectively, both P<0.01); while there was a positive correlation between the vasohibin-1 and the MACC1 (r=0.518, P<0.01). Kaplan-Meier survival analysis showed that the over-expression of vasohibin-1, MACC1 and the low-expression of KAI1 protein were related to the survival rates (all P<0.05). Multi-factor analysis showed that the expression of vasohibin-1, KAI1 protein and the FIGO stage were independent prognosis factors for radical operation of serous ovarian cancer (RR=2.185, 3.893, 0.413; 95% CI=1.263-3.779, 2.190-6.921, 0.251-0.681; all P<0.05).
 Conclusion: The up-regulation of vasohibin-1, MACC1 and down-regulation of KAI1 in serous ovarian cancer are related to the tumor differentiation, clinical stage, metastasis and prognosis. Combined detection of these indexes is useful in predicting the progression and prognosis of serous ovarian cancer.