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Based on diffusion kinetics and cell signal transduction kinetics,we studied the cell cycle arrest characteristics of small drug molecules. The theoretical model takes into account the dynamic characteristics of the transport of small drug molecules across the cell membrane and the blocking effect of drug molecules entering the cell on the cell cycle. We found that the drug molecules crossing the inner layer of the cell membrane and entering the cells will largely determine the blocking effect of drug molecules on related targeted genes and proteins.The transport characteristics of cell membrane to drug molecules are the key factors that affect drug molecules to block cell cycle. In addition,the degree of degradation of drug molecules will change the interaction time between drug molecules and targeted genes and proteins,thereby changing the inhibitory effect on the growth and proliferation of related cells. By the sensitivity analysis of the parameters in the model,we confirmed the inhibitory effect of many factors in the process of drug molecules passing through the cell membrane and entering the cell on the cell cycle. The theoretical results in this paper are consistent with the simulation and experimental observation,which further reveals the physical mechanism of drug small molecules blocking cell cycle,and can provide necessary references and new solutions for designing precise therapeutic drugs. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Prevention & Treatment For Stomatological Diseases is the property of Journal of Prevention & Treatment For Stomatological Diseases Editorial Office 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 discuss the effect of cell division cycle protein 20 (CDC20) on the proliferation and cell cycle of endometrial carcinoma (EC) cells, and to clarify its mechanism. Methods: Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting methods were used to detect the expression levels of CDC20 mRNA and protein in human endometrial stromal T-HESC cell and EC cells (KLE, RL95-2, ZJB-ENC1, and ECC-1 cells). The RL95-2 cells were selected for the subsequent experiments. CDC20 shRNA interference lentivirus was transfected into the RL95-2 cells and the cells were divided into control group, sh-NC group (infected with negative control lentivirus), sh-CDC20 group (infected with CDC20 shRNA interference lentivirus), sh-NC+SM04690 group (infected with negative control lentivirus followed by treatment with 64 nmol·L-1 Wnt/ β -catenin signaling pathway inhibitor SM04690 for 48 h), and sh-CDC20+SM04690 group (infected with CDC20 shRNA interference lentivirus followed by treatment with 64 nmol·L-1 SM04690 for 48 h). RT-qPCR and Western blotting methods were used to detect the expression levels of CDC20 mRNA and proteins in the cells in various groups; CCK-8 method was used to detect the proliferation activities of the RL95-2 cells in various groups; BrdU assay was used to detect the percentages of BrdU positive cells in various groups; flow cytometry was used to detect the percentages of the cells at G2/M stage in various groups; Western blotting method was used to detect the expression levels of β-catenin, oncogene c-Myc, and cyclin D1 proteins in the cells in various groups. Results: Compared with T-HESC cells, the expression levels of CDC20 mRNA and protein in the KLE, RL95-2, ZJB-ENC1, and ECC-1 cells were significantly increased (P<0. 05), and the highest expression levels of CDC20 mRNA and protein were observed in RL95-2 cells. Compared with sh-NC group, the proliferation activities and percentages of the BrdU positive cells in sh-CDC20 group and sh-NC+SM04690 group were significantly decreased (P<0. 05), the percentages of the cells at G2/M phase were significantly increased (P<0. 05), and the expression levels of β -catenin, c-Myc, and cyclin D1 proteins were significantly decreased (P<0. 05). Compared with sh-CDC20 group, the proliferation activity and percentage of BrdU positive cells in sh-CDC20+SM04690 group were significantly decreased (P<0. 05), the percentage of the cells at G2/M phase was significantly increased (P<0. 05), and the expression levels of β-catenin, c-Myc, and cyclin D1 proteins in the cells were significantly decreased (P< 0. 05). Conclusion: CDC20 is highly expressed in the EC cells. Silencing CDC20 may inhibit the cell proliferation by inducing G2/M phase arrest in the RL95-2 cells through the regulation of Wnt/β -catenin signal transduction. [ABSTRACT FROM AUTHOR]

Cell cycle analysis is essential for determining the cell proliferation state, studying cell functions, and evaluating drug effects. Flow cytometry is a commonly used method for cell cycle detection, with propidium iodide (PI) being the most widely used fluorescein. Nevertheless, various factors may affect the test results. Additionally, comparing distributions of immune cell subpopulations across different cell cycle stages can provide valuable insights into immunological responses and disease conditions. In this research, the B16-F10 cell line was used to study the impact of three factors on PI staining-based cell cycle detection: fixation settings, sample preparation conditions, and software analysis. To fix cells, it is suggested to suspend 3 × 106 cells in 300 µL of pre-cooled PBS, add 700 µL of 100% ethanol drop-wise, fix overnight at 4°C or -20°C, and collect at a low flow rate (400-600 events/s) to ensure collection of at least 3 000 singlets. Furthermore, dual-labeling with 5-ethynyl-2'-deoxyuridine (EdU) and PI can accurately distinguish cell cycle phases. And various immune cell subpopulations can be analyzed without cell sorting by combining surface marker staining with PI and Ki-67 staining. Here we review factors affecting cell cycle identification using the PI staining method and provide a standard operating protocol for the experiment. We established the method to combine EdU with PI for cell cycle detection and analysis of immune cell subpopulations, thus expanding the approaches for cell cycle detection. [ABSTRACT FROM AUTHOR]

AIM: To investigate the effects of Xiaozhong-Zhitong mixture (XZZT) on M2 polarization and Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway in mouse microglia (BV2 cells). METHODS: The BV2 cells were divided into 5 groups: blank group, model group [lipo-polysaccharide (LPS)+hypoxia], TAK-242 (resatorvid, a TLR4 inhibitor) group (LPS+hypoxia+TAK-242), XZZT group (LPS+hypoxia+XZZT), and TAK-242+XZZT group (LPS+hypoxia+TAK-242+XZZT). Flow cytometry was used to detect early apoptosis and cell cycle of BV2 cells, and immunofluorescence staining was employed to detect the positive expression of Ml-type marker inducible nitric oxide synthase (iNOS) and M2-type marker CD206. Western blot was utilized to detect the expression of TLR4/MyD88/NF-κB signaling pathway-related proteins, including TLR4, MyD88, NF-κB p65, phosphorylated p65 (p-p65), phosphorylated transforming growth factor-β-activated kinase 1 (p-TAK1), and phosphorylated IκB kinase α/β (p-IKKα/β). RT-qPCR was used to detect the mRNA expression of interleukin-1β (IL-1β), IL-10, tumor necrosis factor-α (TNF-α), TLR4, MyD88, and NF-κB p65. RESULTS: Compared with model group, the rate of early apoptosis was significantly decreased in XZZT group (P<0. 01), the percentage of cells arrested in the S phase was significantly increased (P<0. 01), and the protein levels of TLR4, MyD88, NF-κB p65, p-IKKα/β, p-p65, and p-TAK1 were significantly decreased (P<0. 05 or P<0. 01). Additionally, IL-1β, TNF-α, TLR4, MyD88 and NF-κB p65 mRNA expression levels were significantly decreased (P<0. 05 or P<0. 01), while IL-10 mRNA expression was significantly increased (P<0. 05). Compared with TAK-242 group, the average percentage of iNOS positive area was significantly decreased, while CD206 was significantly increased in TAK-242+XZZT group (P<0. 01). CONCLUSION: The XZZT has the effect of inducing M2 polarization of mouse microglia, and the mechanism may be linked to the inhibition of TLR4/ MyD88/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of kaempferol (KAE) on the function of drug-resistant Bel-7402/5-Fu cells. Methods Bel-7402/5-Fu cells were treated with KAE, and cells were divided into the control group and the drug group (0.064, 0.320, 1.600, 8, 40, 200 µmol/L KAE). Cells were divided into the si-NC group and the DNA-PKcs interference group, or the control group, the KAE group, the KAE+si-DNA-PKcs group or the KAE+DMSO group, the KAE+ MG132 group and the KAE+CQ group based on interfering DNA dependent kinase catalytic subunits (DNA-PKcs) or addition of proteasome inhibitor MG132 or autophagy inhibitor CQ. Cell proliferation was detected using CCK-8. The expression level of histone H2AX phosphorylation (γ -H2AX), DNA-PKcs, DNA double strand break repair/V(D)J recombinant protein (Artemis) and drug pump gene (P-gp) were analyzed using real-time fluorescence quantitative PCR (RT-qPCR) and Western blot assay. Cell cycle and apoptosis were detected by flow cytometry. The stability of DNA-PKcs proteins was analyzed by protein stability experiments. Ubiquitination of DNA-PKcs protein was evaluated by immunoprecipitation assay. Results Compared to the control group, treating cells with 8 µmol/L KAE for 24 h inhibited about 50% of cell proliferation ability. Therefore, this time and concentration were chosen for subsequent research. Compared to the control group, the expression level of γ-H2AX mRNA and protein significantly increased, while expression levels of DNA-PKcs, Artemis and P-gp mRNA and proteins significantly decreased in the KAE group (P＜0.05). Compared to the control group, KAE promoted cell cycle arrest in the G2/M phase of Bel-7402/5-Fu cells and increased cell apoptosis. Compared to the si-NC group, siRNA-1664 significantly downregulated the mRNA and protein expression levels of DNAPKcs (P＜0.05). Compared with the KAE group, the effect of KAE was further promoted in the KAE+si-DNA-PKcs group of Bel-7402/5-Fu cells. Compared with the control group, the protein expression level of DNA-PKcs decreased in the KAE+ DMSO group (P＜0.05). Compared with the KAE+DMSO group, the protein expression level of DNA-PKcs increased in the KAE+MG132 group (P＜0.05), while there was no significant change in the protein expression level of DNA-PKcs in the KAE+CQ group (P＞0.05). Compared to the control group, there was promoted ubiquitination of DNA-PKcs in the KAE+ DMSO group, and the inhibited ubiquitination in the KAE+MG132 group (P＜0.05). Conclusion KAE may induce cell apoptosis and cell cycle arrest in drug-resistant Bel-7402/5-Fu cells. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Shenyang Pharmaceutical University is the property of Shenyang Pharmaceutical University 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 investigate the effect of calcium ions on the expression of kallikrein-4 (KLK4) and cell growth of ameloblast, and to provide an experimental basis for calcium ion promoting normal mineralization of enamel. Methods ALC cells were treated with 0, 2.0, 2.5, 3.0, and 3.5 mmol/L CaCl2 for 24 and 48 h. KLK4 expression was analyzed by qRT-PCR and Western blot analysis. The viability of ALC cells was determined by using CCK-8. AnnexinV-FITC/PI dual staining combined with flow cytometry and Hoechst 33342 staining were used to detect the ALC cell cycle and cell apoptosis. The protein expression level of glucose-regulated protein 78 (GRP78) was measured by Western blot analysis. Results After 24 h of treatment with 2.5, 3.0, and 3.5 mmol/L CaCl2, the expression of KLK4 mRNA was increased (P2, the expression of KLK4 protein was increased (P2, the expression of KLK4 mRNA and protein was increased (P2 (P2. Hoechst 33342 staining results showed that 3.0 mmol/L and 3.5 mmol/L CaCl2 may promote apoptosis in ALC cells. Flow cytometry showed that the proportion of G2/M phase cells and the apoptosis rate increased after 3.5 mmol /L CaCl2 treatment for 24 h (P2 groups. After 24 h of treatment with 3.0 mmol/L and 3.5 mmol/L CaCl2, the expression of GRP78 protein was reduced (P2, the expression of GRP78 protein was reduced (P

BACKGROUND: MiR-338-3p could inhibit osteoclast differentiation, and downregulation of receptor activator of nuclear factor-κB ligand level could promote bone formation. However, it is unclear whether miR-338-3p can affect the proliferation and apoptosis of alveolar bone osteoblasts by regulating the receptor activator of nuclear factor-κB ligand level. OBJECTIVE: To explore the effect and mechanism of miR-338-3p on proliferation and apoptosis of alveolar bone osteoblasts by targeting receptor activator of nuclear factor-κB ligand. METHODS: Human alveolar bone osteoblasts were isolated, transfected and treated with Wnt-C59 (Wnt/β-catenin pathway inhibitor), and divided into transfection control group, miR-338-3p group, miR-338-3p+control group, miR-338-3p+receptor activator of nuclear factor-κB ligand group and miR-338-3p+ Wnt-C59 group. The dual luciferase report experiment was used to verify the regulatory effect of miR-338-3p on receptor activator of nuclear factor-κB ligand. Cell counting kit-8 and 5-Ethynyl-2’-deoxyuridine staining were used to detect cell proliferation levels. Flow cytometry was used to detect cell cycle and apoptosis levels. RT-qPCR was used to detect miR-338-3p, receptor activator of nuclear factor-κB ligand, Wnt-3a, β-Catenin, glycogen synthase kinase-3β mRNA levels. Western blot was used to detect RANKL, proliferating cell nuclear antigen, Ki67, CyclinD1, B-cell lymphoma/leukemia-2, B-cell lymphoma-2 related X protein, Caspase3, Wnt-3a, β-catenin, glycogen synthase kinase-3β protein levels. RESULTS AND CONCLUSION: miR-338-3p could target the regulation of receptor activator of nuclear factor-κB ligand. After overexpression of miR-338-3p, cell survival rate, 5-Ethynyl-2’-deoxyuridine positive cell rate, proportion of S-phase cells were increased, and apoptosis rate was decreased. The mRNA and protein levels of miR-338-3p, proliferating cell nuclear antigen, Ki67, CyclinD1, B-cell lymphoma/leukemia-2, Wnt-3a, and β-catenin were increased, while the mRNA and protein levels of B-cell lymphoma-2 related X protein, Caspase3 protein, receptor activator of nuclear factor-κB ligand, and glycogen synthase kinase-3β were decreased (all P < 0.05). Overexpression of receptor activator of nuclear factor-κB ligand or Wnt-C59 could weaken the effects of overexpression of miR- 338-3p on cell proliferation and apoptosis (all P < 0.05). Overall, miR-338-3p promotes alveolar bone osteoblast proliferation and inhibits apoptosis by targeting receptor activator of nuclear factor-κB ligand, which may act through activation of the Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Objective·To examine the expression level of protein arginine methyltransferase 6 (PRMT6) in breast carcinoma tissues and to assess its impact on the proliferative and migratory behaviors of breast cancer cells.Methods·The PRMT6 transcriptome sequencing data between 33 tumor tissues and normal tissues from The Cancer Genome Atlas (TCGA) database was analyzed through the R language. The gene expression profile interactive analysis (GEPIA2) online database was used to analyze the difference of PRMT6 expression in normal breast tissues and breast cancer tissues. By using the immunohistochemistry (IHC) data of human normal breast tissues and breast cancer tissues from Human Protein Atlas (HPA) database to analyze the protein expression of PRMT6. IHC was used to detect the expression of PRMT6 in breast cancer tissues and paired para-tumor tissues from 27 clinical samples. After PRMT6 was knocked down with small interfering RNA (siRNA) in MDA-MB-231 and MCF-7 cells , the expression of PRMT6 was detected by qRT-PCR and Western blotting. The proliferation ability of breast cancer cells was measured with cell counting kit-8 (CCK-8) assay and colony formation assay. The effect of PRMT6 on the migration ability of breast cancer cells was detected by wound healing assay and Transwell assay. By using the RNA-sequence data from GSE210948 of Gene Expression Omnibus (GEO) database, differentially expressed genes were analyzed in control and low expression groups of PRMT6. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed to reveal the signaling pathways associated with PRMT6. Cell cycle analysis was detected by flow cytometry. The expressions of cyclin D1 and EMT-related proteins (E-cadherin, N-cadherin and Vimentin) were detected by Western blotting after knocking down PRMT6.Results·Bioinformatics analysis and IHC results showed that PRMT6 was highly expressed in breast cancer tissues compared with normal tissues (P=0.000) and para-tumor tissues (P=0.001). qRT-PCR and Western blotting results verified that the siRNA significantly reduced the expression level of PRMT6 in MDA-MB-231 and MCF-7 cell lines compared with the control group (mRNA: P=0.006, P=0.004; P=0.001, P=0.043. Protein: P=0.035, P=0.001; P=0.003, P=0.002). After knocking down PRMT6, the proliferation (P=0.014, P=0.000; P=0.003, P=0.003) and migration (P=0.000, P=0.000; P=0.000, P=0.002) ability of breast cancer cells were inhibited significantly . The KEGG pathway enrichment analysis showed that the expression of PRMT6 affected the cell cycle pathway. After knocking down PRMT6, the expression of cyclin D1 decreased in protein level (P=0.021, P=0.000; P=0.034, P=0.014) and transcription level (P=0.036, P=0.001; P=0.044, P=0.000). Knock down of PRMT6 increased the number of cells in G0/G1 phase (P=0.000; P=0.003) and decreased the number of cells in G2/M phase of the cell cycle . The expression level of E-cadherin increased (P=0.002, P=0.012; P=0.043, P=0.003), while the expression levels of N-cadherin (P=0.004, P=0.041; P=0.032, P=0.034) and Vimentin (P=0.028, P=0.005; P=0.024, P=0.001) decreased in PRMT6 knockdown cells .Conclusion·PRMT6 is highly expressed in breast cancer, which can promote the proliferation and migration of breast cancer cells.

This study aimed to investigate the anti-tumor effect of sesamolin (SES) on hepatocellular carcinoma (HCC) and its molecular mechanism using network pharmacology and experimental verification. The intersecting targets between SES and HCC and their signaling pathways and biological processes were predicted by network pharmacological analysis. The pharmacokinetics properties of SES were analyzed through search against the SwissADME database. The cell count kit-8 (CCK-8) method was used to detect the cell viability. Annexin V-FITC/PI double staining, flow cytometry, Western blotting and cell migration assays were used to analyze the effect and mechanism of SES on cell apoptosis, cell cycle arrest and migration inhibition in Huh-7 cells. The results showed that a total of 64 intersecting targets, 352 gene ontology (GO) functions and 136 Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways were identified by network pharmacological analysis. The in vitro test showed that SES induced mitochondria-dependent apoptosis and G2/M phase arrest in Huh-7 cells through regulating the mitogen-activated protein kinase (MAPK)/signal transducer and activator of transcription-3 (STAT3)/nuclear factor κB (NF-κB) and phosphatidylinositol 3-kinases (PI3K)/protein kinase B (AKT) signaling pathways mediated by reactive oxygen species (ROS). Meanwhile, SES inhibited cell migration through the ROS-mediated PI3K/AKT/glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway. Finally, SwissADME analysis showed that the SES exhibited excellent druglike properties. In conclusion, sesamolin could induce apoptosis, cell cycle arrest and inhibit migration in Huh-7 cells through the ROS-mediated signaling pathways.

Objective·To analyze the expression of cancer-testis antigen (CTA) family member SPANXB (sperm protein associated with the nucleus on the X chromosome B) in liver cancer and its correlation with the prognosis of liver cancer patients, and to explore the impact of SPANXB on liver cancer cell proliferation and its potential mechanism.Methods·By using liver cancer sample data from the cancer genome atlas (TCGA) database, the expression of SPANXB in liver cancer tissue and its correlation with patient survival were analyzed. By constructing stable knockdown of SPANXB and stable overexpression of SPANXB in liver cancer cell lines, the effects of SPANXB on liver cancer cell proliferation were evaluated with live cell imaging experiments, EdU cell proliferation experiments and plate clone formation experiments. The regulatory pathways of SPANXB in liver cancer cell proliferation were explored through RNA-sequence (RNA-seq), and the effect of SPANXB on liver cancer cell cycle was validated through cell cycle experiments. Immunoprecipitation-mass spectrometry (IP-MS) was used to explore the proteins that interacted with SPANXB, and co-immunoprecipitation (Co-IP) was used to verify their interaction.Results·The expression of SPANXB mRNA in liver cancer tissues was higher than that in normal tissues (P=0.003), and was negatively correlated with the survival of liver cancer patients. Stable knockdown of SPANXB could reduce the proliferation and clone formation ability of liver cancer cells, while stable overexpression of SPANXB could promote these processes. The analysis results of RNA-seq showed that knockdown of SPANXB could lead to downregulation of DNA replication and G1/S cell cycle transition-related pathways. The results of cell cycle experiments showed that knockdown of SPANXB could result in changes in the liver cancer cell cycle. The results of IP-MS and Co-IP showed that SPAXNB interacted with cell cycle-related proteins such as mitotic arrest defect 2-like protein 1 (MAD2L1) and WD repeat domain 5 (WDR5).Conclusion·The high expression of SPANXB is negatively correlated with the prognosis of liver cancer. SPANXB may regulate the cell cycle and enhance the proliferation activity of liver cancer cells by interacting with MAD2L1 and WDR5.

Objective To explore the mortality rate and changes in splenic cell cycle and apoptosis in rats with abdominal infection. Methods An animal model of sepsis was induced by cecal ligation and puncture (CLP) in rats. At serial time points 0 h, 6 h, 12 h, 24 h, 48 h, and 72 h after CLP, the animal death rate was calculated. The percentage of cell cycle G1, S, G2/M phase and apoptosis of isolated spleen cells were analyzed using flow cytometer. The expression of p27 of spleen tissue was determined by Western blot analysis. Results In the rat model of CLP-induced systemic inflammatory response, the mortality of animals gradually increased at 0 h, 6 h, 12 h, 24 h, 48 h and 72 h after CLP surgery, reaching a maximum of 88.81%. The abdominal infection in the animals gradually worsened at 0 h, 6 h, 12 h, and 24 h after CLP surgery, but localized and gradually alleviated after 48 h and 72 h. Within 48 h after CLP surgery, there was a blockage in the G1 phase of the spleen cell cycle, an increase in the percentage of apoptotic cells, and an elevation in p27 expression. After 48 h, the G1 phase blockage gradually recovered, the number of apoptotic cell decreased, and p27 expression declined. Additionally, within 48 h after CLP surgery, there was a decrease in the percentage of cells in the S and G2/M phases of the spleen cell cycle, but the percentage of cells in these phases gradually increased after 48 h. ConclusionsCell cycle arrest and apoptosis of the spleen cells are potentially contributed to the change of animal mortality after abdominal infection.

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 death-related 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.

Objective To observe the changes in the cell cycle, apoptosis, phagocytic function, and expression of inflammatory factors in human monocyte/macrophage cell line Rho0 constructed from human monocytic leukemia cells (THP-1 cells) with mitochondrial DNA (mtDNA) depletion. Methods We took THP-1 cells, and used ethidium bromide (EB) to remove mtDNA in the cells to construct the new human monocyte/macrophage Rho0. Rho0 cells were treated with LPS (1 mg/mL) or solvent stimulation for 24 h, respectively, which were taken as the Rho0-LPS group and Rho0- solvent group. Untreated THP-1 cells (Control group) were stimulated with LPS (1 mg/mL) or solvent for 24 h respectively, which were taken as the Control-LPS group and Control-solvent group. Flow cytometry was used to detect the proportion of cell cycle in each group after PI staining. The number of apoptotic cells was measured by Annexin V-FITC/PI method. The phagocytosis function of cells in each group was evaluated by Red-Zymosan dye phagocytic test (mean fluorescence intensity). The expression of inflammatory cytokines (IL-1α, IL-1β, TNF-α, IL-6, IL-8, IL-10, IL-12 mRNA) was detected in Rho0 cells (Rho0 group) and control cells (Control group) by RT-qPCR. Results Compared with Control-solvent group, the proportions of cells in the G0-G1 phase, S phase and G2-M phase in the Rho0-solvent group decreased, the number of apoptotic cells increased and the mean fluorescence intensity of phagocytic dyes increased； the number of apoptotic cells and the mean fluorescence intensity increased in the Control-LPS group (all P<0. 05). Compared with Rho0-solvent group, the proportion of cells in the G0-G1 phase decreased and the number of apoptosis increased in the Rho0-LPS group (all P<0. 05). Furthermore, compared with the control group, the mRNA expression levels of IL-1β, TNF-α, IL-8 and IL10 increased in the Rho0 group (all P<0. 05). Conclusion Removal of mtDNA of mononuclear/macrophage can cause the cell cycle arrest of Rho0 cells, promote the apoptosis, and enhance phagocytic function and proinflammatory function. [ABSTRACT FROM AUTHOR]

Objective To observe the effects of exosomes from human colorectal cancer (CRC) cells (HCT116) with overexpression of microRNA 193b-3p (miR-193b-3p) on the proliferation, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs), and to analyze the targeted relationship between miR-193b-3p and p21-activated kinase 4 (PAK4). Methods Serum from CRC patients and exosomes in the supernatant of HCT116 cells were collected. Transmission electron microscopy and nanoparticle tracking analysis were used to observe and identify exosomes. RT-qPCR was performed to detect exosomal miR-193b-3p and PAK4 mRNA. HUVECs were taken and grouped. HUVECs in the PBS group were added with 100 µL of PBS, HUVECs in the HCT116-Exo group were added with 100 µL of HCT116 cell exosomes, and HUVECs in the HCT116-Exo + GW4869 group were first treated with GW4869 (10 µmol/L) for 30 min and then were added with 100 µL of HCT116 cell exosomes. The proliferation, migration and angiogenesis abilities of HUVECs in each group were detected by EdU assay, Transwell assay and tube formation assay. HUVECs in the logarithmic growth phase were taken and divided into the miR-NC group and the miR-193b-3p group. HUVECs in the miRNC group were co-cultured with HCT116 cell exosomes transfected with miRNA negative control (miR-NC), and the miR193b-3p group with HCT116 cell exosomes transfected with miR-193b-3p mimics. The activity, cell cycle, apoptosis and angiogenesis ability of HUVECs in each group were detected by CCK-8 assay, flow cytometry and tube formation assay. The targeted relationship between miR-193b-3p and PAK4 was detected by the dual-luciferase reporter gene system. Results The expression of miR-193b-3p in serum and exosomes of CRC patients increased, while the expression of PAK4 mRNA decreased (both P<0. 05)； exosomes could promote the proliferation, migration, and angiogenesis of HUVECs (all P<0. 05). Overexpression of miR-193b-3p increased the cell viability and angiogenesis, accelerated cell cycle progression, and inhibited the apoptosis of HUVECs (all P<0. 05)； miR-193b-3p inhibited its luciferase activity by directly targeting PAK4 (P<0. 05). Conclusions The expression of miR-193b-3p in exosomes of CRC serum and cancer cells increases, and the proliferation, migration and angiogenesis of HUVECs with the addition of HCT116 exosomes are enhanced. HUVECs added with HCT116 exosomes transfected with miR-193b-3p mimics show increased proliferation, S phase proportion, angiogenesis ability, and decreased G0/G1 phase proportion and apoptosis rate； miR-193b-3p has a targeted relationship with PAK4. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of expression of microRNA-223（miR-223) on cell proliferation, cell cycle, apoptosis and inflammatory reaction of psoriasis and their mechanism. Methods Human immortalized keratinocytes HaCaT were cultured in vitro. The cells were divided into the control group, model group, model+miR-NC group, model+miR-223 inhibitor group. Except for the control group, cells in the other groups were induced to establish psoriasis cell models by M5 method. Real-time quantitative PCR （RT-qPCR） was used to detect the expression of miR-223 in each group. CCK-8 method and flow cytometry were used to detect cell proliferation activity, cell cycle, and apoptosis in each group. ELISA was used to detect the levels of inflammatory factors such as IL-6, IL-1β and TGF-β1 in supernatant. Western blotting was used to detect the expression of phosphatase and tensin homolog deleted on chromosome ten （PTEN) protein. Results Compared with the control group, the expression level of miR-223 significantly increased （P<0. 05), OD values at 48, 72 and 96 h and the proportion of cells in the cell cycle S phase significantly increased （all P<0. 05), the proportion of cells in the cell cycle G0/G1 and apoptosis rate significantly decreased （all P<0. 05), the levels of IL-6, IL-1β and TGF-β1 significantly increased (all P<0. 05), while the expression of PTEN protein decreased in the model group （P<0. 05）. Compared with the model +miR-NC group, the expression level of miR-223 inhibitor group significantly decreased （P<0. 05)，OD values at 48, 72 and 96 h and the proportion of cells in the S phase decreased （P<0. 05), the proportion of cells in the cell cycle G0/G1 and apoptosis rate significantly increased （P<0. 05), the levels of IL-6, IL-1β and TGF-β1 significantly decreased （P<0. 05), and PTEN protein expression increased in the model +miR-223 inhibitor group （P<0. 05）. There were no significant differences in the above indexes between the model group and the model +miR-NC group （all P>0. 05). Conclusions The expression level of miR-223 increases in psoriasis cells. Down-regulating the level of miR-223 can inhibit the proliferation and inflammatory response of psoriasis cells, and promote their apoptosis and block the cell cycle in G0/G1 phase， which may be related to its regulation of PTEN. [ABSTRACT FROM AUTHOR]

Objective: To explore the relationship between the therapeutic effect of Fu Yuan Sheng Bai Fang (FYSBF) on acute myeloid leukemia (AML) and the JAK/STAT signaling pathway and its mechanism. Methods: AML HL-60 cells were divided into control group, FYSBF group, IL-6 group (IL-6 is the activator of JAK/STAT signaling pathway) and FYSBF+IL-6 group. HL-60 cells in control group were cultured in normal RPMI 1640 medium, HL-60 cells in FYSBF group were cultured in medium which was added with a final concentration of 200 μg/m L FYSBF, and HL-60 cells in IL-6 group were cultured in medium which was added with a final concentration of 100 ng/mL IL-6. HL-60 cells in FYSBF+IL-6 group were cultured in medium which were added with final concentration of 200 μg/m L FYSBF and 100 ng/mL IL-6. The cell proliferation was detected by CCK-8 assay and clonal formation assay, cell cycle distribution and apoptosis were detected by flow cytometry, cell invasion and migration were detected by Transwell assay, and protein expression levels were analyzed by Western blot assay. Results: Compared with the control group, the relative vitality, number of clones, number of migration and invasion cells and proportion of S-phase cells in FYSBF group were decreased (P＜0.05), and the protein expressions of CDK2, cyclin E, N-cadherin and Vimentin were decreased (P＜0.05), the phosphorylation levels of JAK2 and STAT3proteins were decreased (P＜0.05), the cells proportion of G0/G1 phase and apoptosis rate were increased (P＜0.05), and the protein expression of p21 and E-cadherin were increased (P＜0.05) . Compared with the control group, the relative vitality, number of clones, number of migration and invasion cells and proportion of S-phase cells in IL-6 group were increased (P＜0.05), and the protein expressions of CDK2, cyclin E, N-cadherin and Vimentin were increased (P＜0.05), the phosphorylation levels of JAK2 and STAT3proteins were increased (P＜0.05), the cells proportion of G0/G1 phase and apoptosis rate were decreased (P＜0.05), and the protein expression of p21 and E-cadherin were decreased (P＜0.05) . Compared with the IL-6 group, the relative vitality, number of clones, number of migration and invasion cells and proportion of S-phase cells in FYSBF+IL-6 group were decreased (P＜0.05), and the protein expressions of CDK2, cyclin E, N-cadherin and Vimentin were decreased (P＜0.05), the phosphorylation levels of JAK2 and STAT3proteins were decreased (P＜0.05), the cells proportion of G0/G1 phase and apoptosis rate were increased (P＜0.05), and the protein expression of p21 and E-cadherin were increased (P＜0.05) . Conclusion: FYSBF can inhibit the proliferation, migration, invasion and epithelial-mesenchymal transformation of acute myeloid leukemia cells, and promote cell apoptosis and cell cycle arrest, which may be related to inhibiting the activation of JAK2/STAT3 pathway. [ABSTRACT FROM AUTHOR]

Objective To examine the expression level of protein arginine methyltransferase 6 (PRMT6) in breast carcinoma tissues and to assess its impact on the proliferative and migratory behaviors of breast cancer cells. Methods • The PRMT6 transcriptome sequencing data between 33 tumor tissues and normal tissues from The Cancer Genome Atlas (TCGA) database was analyzed through the R language. The gene expression profile interactive analysis (GEPIA2) online database was used to analyze the difference of PRMT6 expression in normal breast tissues and breast cancer tissues. By using the immunohistochemistry (IHC) data of human normal breast tissues and breast cancer tissues from Human Protein Atlas (HPA) database to analyze the protein expression of PRMT6. IHC was used to detect the expression of PRMT6 in breast cancer tissues and paired para-tumor tissues from 27 clinical samples. After PRMT6 was knocked down with small interfering RNA (siRNA) in MDA-MB-231 and MCF-7 cells, the expression of PRMT6 was detected by qRT-PCR and Western blotting. The proliferation ability of breast cancer cells was measured with cell counting kit-8 (CCK-8) assay and colony formation assay. The effect of PRMT6 on the migration ability of breast cancer cells was detected by wound healing assay and Transwell assay. By using the RNA-sequence data from GSE210948 of Gene Expression Omnibus (GEO) database, differentially expressed genes were analyzed in control and low expression groups of PRMT6. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed to reveal the signaling pathways associated with PRMT6. Cell cycle analysis was detected by flow cytometry. The expressions of cyclin D1 and EMT-related proteins (E-cadherin, N-cadherin and Vimentin) were detected by Western blotting after knocking down PRMT6. Results • Bioinformatics analysis and IHC results showed that PRMT6 was highly expressed in breast cancer tissues compared with normal tissues (P=0.000) and para-tumor tissues (P=0.001). qRT-PCR and Western blotting results verified that the siRNA significantly reduced the expression level of PRMT6 in MDA-MB-231 and MCF-7 cell lines compared with the control group (mRNA: P=0.006, P=0.004; P= 0.001, P=0.043. Protein: P=0.035, P=0.001; P=0.003, P=0.002). After knocking down PRMT6, the proliferation (P=0.014, P=0.000; P=0.003, P=0.003) and migration (P=0.000, P=0.000; P=0.000, P=0.002) ability of breast cancer cells were inhibited significantly. The KEGG pathway enrichment analysis showed that the expression of PRMT6 affected the cell cycle pathway. After knocking down PRMT6, the expression of cyclin D1 decreased in protein level (P=0.021, P=0.000; P=0.034, P=0.014) and transcription level (P=0.036, P=0.001; P=0.044, P=0.000). Knock down of PRMT6 increased the number of cells in G0/G1 phase (P=0.000; P=0.003) and decreased the number of cells in G2/M phase of the cell cycle. The expression level of E-cadherin increased (P=0.002, P= 0.012; P=0.043, P=0.003), while the expression levels of N-cadherin (P=0.004, P=0.041; P=0.032, P=0.034) and Vimentin (P= 0.028, P=0.005; P=0.024, P=0.001) decreased in PRMT6 knockdown cells. Conclusion • DRMT6 is highly expressed in breast cancer, which can promote the proliferation and migration of breast cancer cells. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science 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.)

Objective: To discuss the effect of oridonin on the proliferation, migration, epithelial-mesenchymal transition (EMT), and apoptosis of the human nasopharyngeal carcinoma HONE-1 cells, and to clarify its related antitumor mechanism. Methods: The HONE-1 cells were treated with different concentrations (0, 5, 10, 20, 40, 80, and 160 mg·L-1 ) of oridonin for 48 h. CCK-8 method was used to detect the inhibitory rates of proliferation of the cells in various groups and the drug concentration for subsequent experiment was confirmed. The HONE-1 cells were divided into control group, 3 mg·L-1 oridonin group, and 6 mg·L-1 oridonin group. After 24 and 48 h of culture, CCK-8 method was used to detect the proliferation activities of the cells in various groups;5-ethynyl-2'-deoxyuridine (EdU) method was used to detect the rates of EdU-positive cells in various groups; colony formation assay was used to detect the numbers of clone formation in the cells in various groups; Transwell chamber experiment and cell wound assay were used to detect the numbers of migration cells and the scratch healing rates of the cells in various groups; real-time fluorescence quantitative PCR (RT-qPCR) method was used to detect the expression levels of cyclin-dependent kinase 1(CDK1) and cyclin-dependent kinase 4(CDK4) mRNA in the cells in various groups; Western blotting method was used to detect the expression levels of E-cadherin, Vimentin, Caspase-3, and poly ADP-ribose polymerase 1 (PARP1) proteins in the cells in various groups. Results: The CCK-8 method results showed that the half-maximal inhibitory concentration (IC50) of oridonin at 48 h was 12. 18 mg·L-1, and 1/4 IC50 and 1/2 IC50 values were used as the concentrations for subsequent experiments. Compared with control group, after treated for 24 and 48 h, the proliferation activities of the cells in 3 and 6 mg·L-1 oridonin groups were decreased (P<0. 05 or P< 0. 01), the rate of EdU-positive cells were decreased (P<0. 05 or P<0. 01), the numbers of clone formation and migraton cells were decreased (P<0. 05 or P<0. 01), the scratch healing rates were decreased (P<0. 05 or P<0. 01), the expression levels of CDK1 and CDK4 mRNA in the cells were decreased (P< 0. 05 or P<0. 01), the expression levels of E-cadherin, Caspase-3, and PARP1 proteins were increased (P<0. 05 or P<0. 01), and the expression levels of Vimentin protein were decreased (P<0. 05). Conclusion: Oridonin can inhibit the proliferation, clone formation, and migration of the human nasopharyngeal carcinoma HONE-1 cells by downregulating the expression of cell cycle-related proteins and EMT, and promote the apoptosis to exert an antitumor effect. [ABSTRACT FROM AUTHOR]

Objective: Study the function of Nucleoside diphosphate kinase A (NME1) in hepatocellular carcinoma (HCC) cell line HCCLM3 and the regulated pHis substrates of NME1. Methods: The wound healing assay, Transwell chamber assay, and proteomics and phosphoproteomics analysis methods were used to analyze the phenotype and molecular changes of cells after NME1 knockout. Results: Herein, our results showed that histidine kinase NME1 depletion in HCC cell lines inhibited cell motility. Moreover, the proteome analysis found that the differentially expressed proteins after NME1 knockout were primarily involved in the regulation of cell adhesion, cell cycle, and regulation of protein kinase activity. Western blot analysis shows that knockout of NME1 decreases the overall level of pHis modification. Subsequently, using the strategy for unbiased histidine phosphoproteome profiling of dimethyl labeling, Strong cation exchange (SCX) separation of modified and non-modified naked peptides, Cu-iminodiacetic acid (Cu-IDA) enrichment, and mass spectrometry detection, we identified a total of 242 pHis-peptides, corresponding to 206 proteins in the HCC cell line. About 25 % of the pHis-modified proteins were reported for the first time, among which about 30 proteins with significantly down-regulated pHis modification levels in response to NME1 knockout include Cell adhesion molecule 3 (CADM3), Cytochrome C (CYCS), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Phosphofructokinase (PFKM), etc. Conclusion: These findings suggest that NME1 can influence the phenotype changes of tumor cells, such as cell motility, and the dysregulation of the intracellular pHis modification may be widely related to disease procession. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of isoliensinine (Iso) on the proliferation, apoptosis and autophagy of colon cancer SW480 cells through PI3K/Akt/mTOR signaling pathway. Methods: Colon cancer SW480 cells were treated with 10, 20 and 40 μmol/L Iso, and the effects of Iso on the cell proliferation capacity, apoptosis and expressions of autophagy related proteins LC3 I, LC3II and p62 were detected by CCK-8, flow cytometry and Western blot, respectively. Then, SW480 cells were treated respectively with 20 μmol/L Iso and 25 μmol/L PI3K activator 740 Y-P, and the cells were divided into the control group, the 740 Y-P group, the Iso group and the Iso+740 Y-P group. The effects of 740 Y-P on the apoptosis and the expressions of LC3 I, LC3 II, p62, PI3K, p-PI3K, mTOR and p-mTOR proteins in each group were detected by flow cytometry and WB. Results: After treatments with 10, 20 and 40 μmol/L Iso, the proliferation capacity of SW480 cells was decreased significantly (all P<0.05); the apoptosis rate was increased significantly (all P<0.05); the expressions of LC3 II/LC3 I were up-regulated significantly (all P<0.05), and the expression of p26 protein was down-regulated significantly (all P<0.05). After treatments with Iso and 740 Y-P, compared with the control group, the apoptosis rate and LC3 II/LC3 I expression of the 740 Y-P group were decreased significantly (both P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were increased significantly (all P<0.05).The apoptosis rate and LC3 II/LC3 expression in the Iso group were increased (both P<0.05) and the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the 740 Y-P group, the apoptosis rate and LC3 II/LC3 I expression were increased in the Iso+740 Y-P group (P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the Iso group, the apoptosis rate and LC3 II/LC3 I expression were decreased (both P<0.05), and the expressions of p26, p-PI3K/PI3K, and p-mTOR/mTOR were increased significantly (all P<0.05) in the Iso+740 Y-P group. Conclusion: Iso inhibits the proliferation and induces the apoptosis and autophagy of SW480 cells by inhibiting PI3K/Akt/mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

Copyright of Traditional Chinese Drug Research & Clinical Pharmacology is the property of Traditional Chinese Drug Research & Clinical Pharmacology Editorial Office 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 observe the effects of long non-coding RNA (IncRNA) nuclear-enriched abundant transcript 1 (NEAT1) on the proliferation and apoptosis of human laryngeal squamous cell carcinoma (LSCC) cells, as well as its targeted relationship with microRNA-214 (miR-214). Methods Real-time fluorescence quantitative PCR (RT-qP-CR) was used to detect IncRNA NEATI and miR-214 in immortalized human epidermal cells Hacat and human LSCC cell lines (FD-LSC-1, Hep-2, and TU177), and then TU177 cells were selected as the experimental cells. TU177 cells in the logarithmic growth phase were divided into groups A and B, which were transfected with si-IncRNA NEATI (knockdown of IncRNA NEATI expression) and si-NC (random meaningless sequence), respectively. At 24 h, cell proliferation was observed using CCK-8 assay, colony formation experiment was conducted to observe clone formation, flow cytometry was used to analyze cell cycle distribution and apoptosis rate, while RT-qPCR was performed to detect IncRNA NEATI and miR-214 expression in both groups. TU177 cells in the logarithmic growth phase were divided into groups I, II, III and IV, which were transfected with WT-IncRNA NEAT1 and miR-214 mimic, WT-IncRNA NEATI and miR-NC, MUT-IncRNA NEAT1 and miR-214 mimic, and MUT-IncRNA NEAT1 and miR-NC, respectively. Th targeted relationship between In-CRNA NEATI and miR-214 was validated by dual luciferase reporter gene assay. Results Compared with the group B, the OD values of TU177 cells were lower at 24, 48, and 72 h of cultivation; at 14 d of cultivation, the number of clone formation in TU177 cells was smaller; the proportion of cells in the Go/G, phase was higher, the proportion of cells in the S phase was lower, and the apoptosis rate was higher in the group A (all P<0.05). Compared with the group B, at 24 h of transfection, the relative expression of IncRNA NEATI was lower and the relative expression of miR-214 in TU177 cells was higher in the group A (both P<0.05). At 48 h of cultivation, the luciferase activity of TU 177 cells in the groups I, II, III and IV was 0.63±0.08, 0.99 ±0.01, 1.02 ± 0.02, and 0.98 ± 0.03, respectively. Compared with the group I, group II had lower cell luciferase activity (P<0.05). Conclusions Knockdown of IncRNA NEATI expression can inhibit the proliferation and cloning of TU177 cells, block the cell cycle in Go/G, phase, and promote ote the apoptosis. LncRNA NEATI is targetedly associated with miR-214 in TU177 cells. LncRNA NEATI may inhibit the proliferation and cloning of TU 177 cells, block the cell cycle in Go/G, phase, and promote the apoptosis by targetedly binding with miR-214. [ABSTRACT FROM AUTHOR]

The cell cycle is the series of events by which a cell divides into two daughter cells and is guided by a unique system of cell cycle regulation. The disorder of cell cycle regulation system is the fundamental cause of tumorigenesis. This paper will focus on the key regulatory points of cell cycle and discuss the research progress of tumor cell cycle regulation, so as to provide theoretical basis for the development of new therapeutic strategies for tumors. [ABSTRACT FROM AUTHOR]

romodomain-containing protein 4 (BRD4), a pivotal member of the bromodomain and extra-terminal domain (BET) family, is integral to the regulation of vital biological processes, including the cell cycle and gene expression. Studies have identified extensive expression of BRD4 in cardiomyocytes and its influence on various signaling pathways. The dysregulation of BRD4 leads to significant alterations in these pathways, culminating in pathological states such as myocardial inflammation, fibrosis, oxidative stress, and hypertrophy. These changes are instrumental in the onset and progression of cardiovascular diseases. This review summarizes the advances in research on BRD4 and its inhibitors in the context of cardiovascular diseases, with a focus on providing insights for precise therapeutic interventions. [ABSTRACT FROM AUTHOR]

: This study examined the inhibitory effect of peiminine on the human colonic adenocarcinoma cell line SW480 and explored the underlying mechanisms. METHODS: SW480 and human normal colonic epithelial CCD-841CoN cells were treated with different concentrations of peiminine and subjected to the CCK-8 assay to select the optimal treatment time and concentration of the compound. SW480 cell migration and invasion were evaluated by the wound-healing and Transwell assays. Cell cycle progression was analyzed by flow cytometry. The expression levels of cell cycle-related proteins were examined by Western blot. SW480 xenograft tumor model was established in nude mice to examine the effect of peiminine on tumor growth and the expression of cell cycle-related proteins in vivo. RESULTS: Peiminine (110 mg/L) significantly inhibited the proliferation of SW480 cells compared with the control group (P<0.01), caused cell cycle arrest at G, phase, and significantly downregulated the expression of cyclin dependent kinase 2(CDK2), DK4, CDK6, cyclin D1, p-Rb/Rb, E2F1, E2F3, and E2F4 (P<0. 05). Peiminine inhibited SW480 xenograft tumor growth, prolonged the survival of model mice, and affected the expression of CDK2, CDK4, CDK6, and cyclin D1 in tumor tissues. CONCLUSION: Peiminine promotes G1 phase arrest by down-regulating the expression of CDK2, CDK4, CDK6, and cyclin D1, thereby inhibiting the proliferation of SW480 cells. [ABSTRACT FROM AUTHOR]

To explore the mortality rate and changes in splenic cell cycle and apoptosis in rats with abdominal infection. Methods An animal model of sepsis was induced by cecal ligation and puncture (CLP) in rats. At serial time points 0 h, 6 h, 12 h, 24 h, 48 h, and 72 h after CLP, the animal death rate was calculated. The percentage of cell cycle G1, S, G2/M phase and apoptosis of isolated spleen cells were analyzed using flow cytometer. The expression of p27 of spleen tissue was determined by Western blot analysis. Results In the rat model of CLP-induced systemic inflammatory response, the mortality of animals gradually increased at 0 h, 6 h, 12 h, 24 h, 48 h and 72 h after CLP surgery, reaching a maximum of 88.81%. The abdominal infection in the animals gradually worsened at 0 h, 6 h, 12 h, and 24 h after CLP surgery, but localized and gradually alleviated after 48 h and 72 h. Within 48 h after CLP surgery, there was a blockage in the G1 phase of the spleen cell cycle, an increase in the percentage of apoptotic cells, and an elevation in p27 expression. After 48 h, the G1 phase blockage gradually recovered, the number of apoptotic cell decreased, and p27 expression declined. Additionally, within 48 h after CLP surgery, there was a decrease in the percentage of cells in the S and G2/M phases of the spleen cell cycle, but the percentage of cells in these phases gradually increased after 48 h. ConclusionsCell cycle arrest and apoptosis of the spleen cells are potentially contributed to the change of animal mortality after abdominal infection. [ABSTRACT FROM AUTHOR]

Fucoxanthin is a carotenoid, reddish-brown in color, which is mainly derived from marine organisms such as brown algae and diatoms, and has aroused extensive scientific interest due to its unique chemical structure and abundant biological activities. Studies have shown that fucoxanthin processes significant anti-inflammatory and anti-cancer activities. Its anti-inflammatory mechanisms primarily include inhibiting oxidative stress, regulating inflammatory factors, inducting cell autophagy, and resisting cell apoptosis, etc. The main mechanism of anti-cancer activity includes inducing cell autophagy and apoptosis mechanisms, regulating the cell cycle, inhibiting cell migration, and suppressing cell invasion, among other aspects. In this paper, the research progress of anti-inflammatory and anti-cancer activities and related mechanisms of action of fucoxanthin is briefly summarized, which provides the theoretical basis and reference for further research and development of fucoxanthin. [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

Cell cycle turnover depends on cyclin-dependent kinase (CDKs). CDK is the protein kinase family that plays the key role in regulating cell cycle and gene transcription. CDK12 and CDK13 perform the essential work in DNA damage response, RNA splicing regulation, transcription, and cell cycle regulation. In recent years, CDK12 and CDK13 are expressed abnormally or mutated in a variety of cancers; thus, they are necessary in cancer development and have become popular topics of research. Based on literature from Google Scholar, PubMed, and WanFang database, aspects of CDK12 and CDK13 such as basic structure, biological function, correlation with malignant tumors, and research progress of targeted inhibitors are summarized to provide the direction for the subsequent research of the new targets and mechanisms for targeting therapy while offering a novel approach for the prevention, diagnosis, and treatment of malignant tumors in the future.

Objective To explore the effect of sorafenib on HeLa cell proliferation by inducing cell apoptosis and autophagy and its impact on drug resistance. Methods The drug-resistant cell strains were constructed through intermittent induction method, with concentrations of 0, 2.5, 5.0, 7.5, 10.0, 15.0, 20.0 μmol/L. HeLa cells were incubated with increasing concentrations of sorafenib with each concentration for 1 week. The drug-resistant cell strains with stable passages were collected. MTT assay was used to detect the effect of sorafenib on cell proliferation. Cell cycle distribution was analyzed by flow cytometry. The change in the expression of drug-resistant and apoptotic genes in the parents and drug-resistant cell strains under different drug concentrations was examined by semi-quantitative PCR. The changes of apoptotic related marker proteins LC3-Ⅰ and LC3-Ⅱ were detected by Westernblot. Results Stable drug-resistant strains were successfully obtained; Drug-treated cells were more blocked in the G1 phase. In drug-resistant cells, the expression of apoptosis suppressor gene Bcl-2 was significantly decreased and the apoptotic gene Bax as well as the drug-resistant genes were all significantly increased(P＜0.05). The LC3-Ⅱ/LC3-Ⅰ ratio of drug-resistant cells was significantly higher than that of parent cells(P＜0.05). Conclusions Sorafenib may block the cell cycle, suppress malignant cell proliferation and promote autophage. On one hand, autophagy participates in the development of cell drug resistance and promotes cell survival. On the other hand, drug-induced autophagy may activate some of apoptotic signaling pathway in drug-resistant cells and promote the reversal of cell drug resistance.

Objective To explore the intrinsic relationship between circadian clock and cell cycle in osteoarthritis(OA) chondrocytes, especially the regulation of cell cycle-related genes by the clock gene Bmal1. Methods The chondroid ATDC5 cells induced by insulin-transfering-selenium(ITS) were divided into control group, OA group and LV-Bmal1 group. The cell viability of each group was detected by CCK8 method. The expression of Bmal1, Per1, Wee1, Cdk1, Ccnb1 and Mmp13 mRNA in each group was detected by RT-qPCR. The expression of BMAL1, PER1, WEE1, CDK1, CCNB1 and MMP13 protein in each group was detected by Western blot. The effects of Bmal1 on different stages of cell cycle and apoptosis was analyzed by flow cytometry. The regulation of Bmal1 on Per1, Wee1, Cdk1, Ccnb1 and Mmp13 and their roles in OA were analyzed. Results Compared with the normal group, the cell viability of the OA group was increased, the relative mRNA expression of Bmal1 and Wee1 in the OA group decreased, and the relative mRNA expression of Per1, Cdk1, Ccnb1 and Mmp13 increased significantly. The cell viability of LV-Bmal1 group decreased, the relative expression of Bmal1 and Wee1 mRNA increased, and the relative expression of Per1, Cdk1, Ccnb1 and Mmp13 mRNA decreased(P＜0.05). Correlation analysis showed that Bmal1 was positively correlated with Wee1 and they were negatively correlated with Per1, Cdk1, Ccnb1 or Mmp13. The results of Western blot showed that protein expression in different groups were consistent with the trend of PCR. The results of cell cycle and apoptosis showed that compared with the normal group, the S phase and G2/M phase of the OA group were shortened, the proportion of cells decreased significantly, and the proportion of early and late apoptosis increased. The S phase and G2/M phase of the LV-Bmal1 group were prolonged, the proportion of cells was increased, and the proportion of early and late apoptosis was decreased. Conclusions Circadian clock gene Bmal1 in inflammatory chondrocytes might regulate the expression of cell cycle-related genes.

Objective To investigate the effect and mechanism of targeted regulation of phosphatase and tensin homolog (PTEN) and phosphatidylinositol 3-kinase (PI3K) on the proliferation and apoptosis of nephroblastoma cells. Methods Posto-perative tumor tissue and normal paracancerous tissue were collected from 15 children with nephroblastoma, and Western blotting and quantitative real-time PCR were used to measure the protein and mRNA expression levels of PTEN and PI3K in tissue. Nephroblastoma SK-NEP-1 cells were divided into control group (group A, transfected with NC siRNA and Flag empty vector), PTEN overexpression group (group B, transfected with NC siRNA and Flag-PTEN expression vector), PI3K knockdown group (group C, transfected siPI3K and Flag empty vector), a nd combined targeting group (group D, transfected siPI3K and FLAG-PTEN). Western blotting was used to measure the expression levels of PI3K, protein kinase A (AKT), and phosphorylated AKT (p-AKT) in SK-NEP-1 cells at 24 h after transfection; CCK-8 assay was used to observe the proliferation of SK-NEP-1 cells at 24, 48, and 72 h of intervention; flow cytometry was used to observe apoptosis rate and cell cycle at 24 h after transfection. Results Compared with normal paracancerous tissue, nephroblastoma tumor tissue showed significant increases in the protein and mRNA expression levels of PI3K (t=22.862,7.098,P

Objective To investigate the effects of aucubin (AU) on the proliferation, apoptosis, and cell cycle of human liver cancer cells line HepG2 and its mechanism of action. Methods HepG2 cells were cultured in vitro, CCK-8 method was applied to screen the optimal dosage concentration of AU. HepG2 cells were randomly grouped into a control group, an AU 12.5 mg/L group (AU L group), an AU 62.5 mg/L group (AU H group), and an AU H+Akt pathway agonist (SC79) group(AU H+SC79 group). The cell proliferation was observed in each group; 5-Ethynyl-2′-deoxyuridine (EDU) method was applied to detect cell proliferation; Flow cytometry was applied to detect cell apoptosis and cell cycle; Western blot was applied to detect the expression levels of phosphorylated protein kinase B (p-Akt), Akt, p-MDM2, MDM2, p-p53, and p53 proteins. Results AU concentrations of 12.5 and 62.5 mg/L were selected for subsequent experiments.Compared with 0 mg/L AU, the proliferation of 12.5 and 62.5 mg/L AU cells was obviously reduced (P＜0.05);Compared with the control group, the number of suspended and exfoliated cells in the AU L and AU H groups gradually increased. Cells shrunk and became round. The proportion of G0/G1 phase cells, the proportion of EDU positive staining cells increased and the expression level of p-Akt/Akt and p-MDM2/MDM2 proteins decreased. The proportions of S and G2/M phase cells, the rate of cell apoptosis, and the expression level of p-p53/p53 protein all increased (P＜0.05). Compared with the AU H group, the above changes in the AU H+SC79 group were recovered(P＜0.05). After AU treatment, the tumor volume and weight of transplanted nude mice decreased. Conclusions AU may inhibit the proliferation of liver cancer cells, induce cell cycle arrest and apoptosis by regulating the Akt/MDM2/p53 signaling pathway.

Objective: To investigate the expression of TFCP2L1 in human endometrial cancer and to analyze the effect of TFCP2L1 on the proliferation and migration of endometrial cancer cells. Methods: (1) The expression level of TFCP2L1 in endometrial cancer and the survival of patients were analyzed using the TCGA and GTEx databases. The expression of TFCP2L1 in normal endometrial epithelial cells and various endometrial cancer cell lines was verified by Western blot. (2) TFCP2L1 was knocked out of the Ishikawa cell line using CRISPR-Cas9 technology, and single cells were selected by flow cytometry for culture to form monoclonal cell lines to study the effect of TFCP2L1 on the cell cycle and cell proliferation of endometrial cancer. The expression of cell cycle-related proteins was detected by Western blot and cell immunofluorescence, and cell proliferation was detected by plate cloning assay and CCK8 assay. (3) The invasion and metastasis ability were detected by Transwell chamber and scratch assay. Results: Analysis of TCGA and GTEx databases revealed that TFCP2L1 was highly expressed in endometrial cancer and was associated with poor prognosis in cancer patients. After knocking out TFCP2L1, the protein levels of Ki67, Cyclin D1, and Cyclin D2 were significantly downregulated. The results of CCK8 and plate cloning experiments showed that knocking out TFCP2L1 could significantly reduce the proliferation ability of endometrial cancer cells. The results of the scratch assay and Transwell invasion assay showed that the invasion and migration abilities of endometrial cancer cells knocked out TFCP2L1 were weakened. Conclusion: This study proves that TFCP21L1 is a carcinogen of endometrial cancer. High expression of TFCP2L1 may be associated with poor prognosis of endometrial cancer. Knocking out TFCP2L1 can inhibit the invasion and metastasis of endometrial cancer. [ABSTRACT FROM AUTHOR]

BACKGROUND: Skin damage caused by radiation therapy and nuclear accidents is still a serious medical problem. It is difficult to achieve effective treatment results with single prevention and treatment methods. It is an important research direction to find new comprehensive treatment methods. OBJECTIVE: To observe the protective effect and the underlying mechanism of 1,2-propanediol combined with hepatocyte growth factor-modified exosomes derived from dental pulp stem cells on human epidermal radiation damage cell models. METHODS: (1) After infection of human dental pulp stem cells using recombinant adenovirus of human hepatocyte growth factor gene, exosomes, i.e., Ad. HGF DPSC-Exo, were isolated with ultracentrifugation. (2) HaCat cells were irradiated with X-ray. The cells were treated with 1,2-propanediol before irradiation and Ad.HGF DPSC-Exo after irradiation. Cell proliferative activity was determined by CCK-8 assay. Cell apoptosis was detected by flow cytometry. Cell migration was detected by cell scratch assay. The expression levels of P21 and P53 were detected by PCR. RESULTS AND CONCLUSION: 1,2-Propanediol, Ad.HGF.DPSC-Exo, Ad.HGF.DPSC-Exo + 1,2-propanediol could significantly improve the growth inhibition of HaCaT cells, reduce cell apoptosis, elevate cell proliferation and migration, and exhibit a good radiation protection effect. Moreover, the combined effect of Ad.HGF.DPSC-Exo + 1,2-propanediol was better. Furthermore, Ad.HGF.DPSC-Exo + 1,2-propanediol alleviated the cellular G2/M phase block and decreased the expression of cell cycle genes P53 and P21. In conclusion, 1,2-propanediol pretreatment combined with Ad.HGF.DPSC-Exo had significant protective effects on radiation-induced HaCaT cell injury and it provided novel ideas and potential methods for the prevention and treatment of radiation-induced skin damage. [ABSTRACT FROM AUTHOR]

Objective To explore the molecular mechanism of Gasdermin B (GSDMB) regulating the fate of intestinal epithelial cells. Methods The human GSDMB plasmid was overexpressed into two human intestinal epithelial cell lines (NCM460 and HT-29 cells) and human colon-derived organoids. Western blotting was used to confirm the efficiency of electroporation. Cell counting kit (CCK8), cell apoptosis, and cell cycle by flow cytometry were performed to analyze the effect of GSDMB overexpression on cell function. Transcriptome sequencing was used to analyze the downstream effector molecules of GSDMB. T test was used to compare the data between the two groups. Results The overexpression of GSDMB protein in the two intestinal epithelial cell lines was successfully reconstructed. The absorbance value (A) of human intestinal epithelial cells overexpressing GSDMB protein [NCM460 cells: (1.17±0.01), HT-29 cells: (0.96±0.06)] was significantly lower than that of blank control cells [NCM460 cells: (1.67±0.12), HT-29 cells: (1.24±0.07)] (t=7.24 and 5.46, P<0.05). The number of apoptotic cells in the GSDMB overexpression group [NCM460 cells: (12.03±1.55), HT-29 cells: (29.30±4.48)] was significantly higher than that in the blank group [NCM460 cells: (4.96±1.74), HT-29 cells: (6.95±3.42)](t=5.26 and 6.97, P<0.05). Cell cycle analysis showed that the ratio of cells at G0/G1 phase in the GSDMB overexpression group [NCM460 cells: (47.98±5.28)%, HT-29 cells: (38.04±3.45)%] was significantly lower than that in the control group [NCM460 cells: (59.54±3.90)%, HT-29 cells: (63.81±1.76) % (t=3.05 and 11.53, P<0.05). Transcriptome sequencing results showed that the dual specificity phosphatase 4 and 6 (DUSP4 and DUSP6) genes were significantly upregulated after GSDMB protein expression. Fluorescence quantitative PCR results confirmed that the relative expression levels of DUSP4 (2.45±0.15) and DUSP6 (4.34±0.22) in intestinal epithelial cells transfected with GSDMB were significantly higher than those in the control group (1.06±0.05 and 1.01±0.02)(t= 15.08 and 26.52, P<0.05). After GSDMB-expressing NCM460 cells were treated with the DUSP inhibitor BCI, the BCI treatment group had a significantly increased expression level of p-ERK compared to the control group [(1.14± 0.17) vs. (0.58±0.12)](t=5.42, P=0.002); the A value (1.84±0.07) and G0/G1 phase ratio (59.83±2.17)% in the BCI treatment group were significantly higher than those in the non-treatment group [(1.52±0.10) and (52.10± 2.23)%, and the number of apoptosis in the BCI treated group (7.60±0.56) was significantly lower than that in the untreated group (12.57±1.00) (t=4.71, 4.31, 7.52, P<0.05). TUNEL staining in human colon organoids showed a significant increase in apoptotic cells, and the relative expression level of DUSP6 protein (0.85±0.09) was significantly higher than that of the control group (0.21±0.04), accompanied by a decrease in p-ERK levels [(0.83± 0.18) vs. (0.19±0.06)], with statistical significance (t=11.95, P<0.001; t 6.56, P<0.001). Conclusion GSDMB may inhibit cell proliferation, induce cell cycle arrest, and promote apoptosis by upregulating dual specificity phosphatase DUSP6-mediated ERK phosphorylation, thus affecting the fate of intestinal epithelial cells. [ABSTRACT FROM AUTHOR]

Cell cycle turnover depends on cyclin-dependent kinase (CDKs). CDK is the protein kinase family that plays the key role in regulating cell cycle and gene transcription. CDK12 and CDK13 perform the essential work in DNA damage response, RNA splicing regulation, transcription, and cell cycle regulation. In recent years, CDK12 and CDK13 are expressed abnormally or mutated in a variety of cancers; thus, they are necessary in cancer development and have become popular topics of research. Based on literature from Google Scholar, PubMed, and WanFang database, aspects of CDK12 and CDK13 such as basic structure, biological function, correlation with malignant tumors, and research progress of targeted inhibitors are summarized to provide the direction for the subsequent research of the new targets and mechanisms for targeting therapy while offering a novel approach for the prevention, diagnosis, and treatment of malignant tumors in the future. [ABSTRACT FROM AUTHOR]

Objective: To discuss the inhibitory effect of Schisandrin B on the proliferation of pancreatic cancer Pan02 cells, and to clarify the mechanism. Methods: CCK-8 method was used to detect the proliferation rates of the Pan02 cells after treated with different concentrations (0, 0. 78, 1. 56, 3. 12, 6. 25, 12. 50, and 25. 00 mg·L-1) of Schisandrin B to select the optimal concentration and treatment time of Schisandrin B. The mouse pancreatic cancer Pan02 cells were divided into control group (0 mg·L-1 Schisandrin B), 2. 5 mg·L-1 Schisandrin B group, 5. 0 mg·L-1 Schisandrin B group, and 10. 0 mg·L-1 Schisandrin B group. The morpholoy of Pan02 cells invarious groups was observed with light microscope; 5-ethynyl-2'-deoxyuridine (EdU) staining assay was used to detect the positive expression rates of the Pan02 cells in various groups; flow cytometry was used to detect the percentages of the Pan02 cells at different cell cycles and the apoptotic rates of the cells in various groups; Western blotting method was used to detect the expression levels of cell cycle and apoptosis-related proteins in the cells in various groups. Results: The CCK-8 method results showed that after treated with Schisandrin B for 48 and 72 h, compared with 0 mg·L-1 Schisandrin B, the proliferation rates of the Pan02 cells after treated with different concentrations of Schisandrin B were decreased (P<0. 01), especially at 72 h. 0. 25, 5. 0, and 10. 0 mg·L-1 Schisandrin B were selected to treat the Pan02 cells, and 72 h was the treatment time. In control group, the Pan02 cells had a spindle shape, with good condition, and grew closely adhered to the wall with normal organelles and cytoplasm, in 2. 5 and 5. 0 mg·L-1 Schisandrin B groups, the cell volume was decreased, the intercellular adhesion was disappeared, and the cell membrane was intact but more permeable; the cytoplasm shrank and vacuolar structures appeared inside the cells, with some fragmented and floating on the surface of the solution; in 10. 0 mg·L-1 Schisandrin B group, the Pan02 cells exhibited notable apoptotic bodies, indicating an apoptotic state. The EdU staining results showed that compared with control group, the rates of EdU positive cells in 2. 5, 5. 0, and 10. 0 mg·L-1 Schisandrin B groups were significantly decreased (P<0. 01). The flow cytometry results showed that compared with control group, the percentages of the cells at S phase in 2. 5, 5. 0, and 10. 0 mg·L-1 Schisandrin B groups were significantly increased (P<0. 01), while the percentages of the cells at G2/M phase were significantly decreased (P<0. 01), and the percentages of the cells at G0/G1 phase in 5. 0 amd 1. 0 mg·L-1 Schisandrin groups were decreased (P<0. 01); compared with control group, the apoptotic rates of the cells in 2. 5, 5. 0, and 10. 0 mg·L-1 Schisandrin B groups were significantly increased (P<0. 01). The Western blotting results showed that compared with control group, the expression levels of p27, B-cell lymphoma 2 (Bcl-2) associated X protein (Bax), cleaved cysteine aspartic acid protease-3 (cleaved Caspase-3), and cleaved poly adenosine diphosphate (ADP) ribose polymerase (cleaved PARP) proteins in the cells in 2. 5 mg·L-1 Schisandrin B group were significantly increased (P<0. 05 or P<0. 01), the expression levels of cyclin A2, cyclin E2, and Bcl-2 proteins in the cells in 5. 0 and 10. 0 mg·L-1 Schisandrin B groups were significantly decreased (P<0. 05 or P<0. 01), while the expression levels of p27, Bax, cleaved Caspase-3, and cleaved PARP proteins in the cells in 5. 0 and 10. 0 mg·L-1 Schisandrin B groups were significantly increased (P<0. 01). Conclusion: Schisandrin B has an inhibitory effect on proliferation of the pancreatic cancer Pan02 cells, and its mechanism may be related to the activation of the cysteine aspartic acid protease-3 (Caspase-3) pathway to induce the apoptosis and activating p27 protein to induce the arrest of cell cycle at S phase [ABSTRACT FROM AUTHOR]

Copyright of Practical Oncology Journal is the property of Journal of Practical Oncology Editorial Office 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.)

AIM: To establish a radioresistant lung adenocarcinoma cell model in vitro and investigate the biological changes related to radiation resistance in tumor cells. METHODS: A radioresistant lung adenocarcinoma cell model namely A549RR, was established by fractionated irradiation. The A549 and A549RR cells were randomly divided into two groups, which were irradiated with 0 and 8 Gy. The proliferation ability of parental cells and radioresistant cells were detected to assess their sensitivity to radiotherapy. The A549, A549RR and A549RR' （partially radioresistant） cell lines were divided into cisplatin chemotherapy group, cisplatin chemotherapy combined with radiotherapy group and blank control group to evaluate the sensitivity to chemotherapy and radiochemotherapy. Flow cytometry was used to detect the changes in cell cycle and apoptosis after irradiation, while the comet assay was used to evaluate DNA damage. The levels of reactive oxygen species （ROS） and 8-hydroxydeoxyguanosine （8-OHdG） levels were measured, and senescence-associated β-galactosidase staining was performed. RESULTS: Compared with parental A549 cells, A549RR cells exhibited enhanced radiation resistance （P<0. 01）, decreased proliferation capacity （P<0. 01）, and no significant difference in cell viability before and after chemotherapy. In comparison to A549 cells, the proportion of A549RR cells at G2/M phase was significantly decreased （P<0. 01）, and that at S phase was significantly increased （P<0. 01）. After exposed to high-dose irradiation, A549 cells displayed S phase arrest, whereas A549RR cells showed no significant difference in cell cycle before and after irradiation. Additionally, A549RR cells exhibited reduced DNA damage （P<0. 01） and lower levels of ROS （P<0. 01） compared with A549 cells following irradiation. There were no significant differences in the levels of 8-OHdG and apoptotic rate before and after irradiation in A549RR cells. CONCLUSION: In this study, by establishing a radiation-resistant cell model, the impact of radiotherapy on the survival of radioresistant lung cancer cells was investigated, as well as the corresponding changes in cell cycle progression, DNA damage, apoptosis, senescence, and oxidative stress levels. These findings are expected to contribute to a better understanding of the mechanisms underlying radiation resistance in lung cancer and may inform further research in this field. [ABSTRACT FROM AUTHOR]

AIM: To explore the role and mechanisms of Obg-like ATPase 1（ OLA1） gene in the growth of human B-cell lymphoma Romas cells. METHODS: The Gene Expression Profiling Interactive Analysis （GEPIA） database was utilized to analyze the differential expression of OLA1 in diffuse large B-cell lymphoma （DLBCL） tissues, and the impact of OLA1 expression on the survival rate of patients was examined by the GSE181063 database. The stable cell lines with knockdown of the OLA1 gene（ OLA1-sh） and negative control were established by Romas cells via infection with retrovirus. The growth of Romas cells were assessed by cell counting, CCK-8 assay, subcutaneous tumor experiment in mice and Ki67 immunohistochemical staining. Flow cytometry was employed to investigate the cell cycle and apoptosis. TUNEL fluorescence staining was conducted to evaluate the apoptosis. The relationship between OLA1 gene and cell cycle/apoptosis-related genes was analyzed by differential genes from 48 DLBCL patients in The Cancer Genome Atlas （TCGA） database. RT-qPCR was utilized to detect the cell cycle- and apoptosis-related genes. Western blot analysis was performed to detect the cell cycle- and apoptosis-related proteins. RESULTS: The OLA1 gene was highly expressed in DLBCL （P< 0. 05）, and OLA1 knockdown significantly inhibited Romas cell proliferation, arrested the cell cycle, and induced apoptosis （P<0. 05 or P<0. 01）. Additionally, OLA1 knockdown promoted the expression of Bax, and suppressed cyclin-dependent kinase 6 （CDK6）, cyclin E1 and BCL2 expression （P<0. 05 or P<0. 01）. CONCLUSION: Knockdown of OLA1 markedly suppresses the growth of human B-cell lymphoma Romas cells. The underlying mechanism might involve the blockade of cell cycle and the induction of apoptosis. [ABSTRACT FROM AUTHOR]

Objective To explore the intrinsic relationship between circadian clock and cell cycle in osteoarthritis(OA) chondrocytes, especially the regulation of cell cycle-related genes by the clock gene Bmal1. Methods The chondroid ATDC5 cells induced by insulin-transfering-selenium(ITS) were divided into control group, OA group and LV-Bmal1 group. The cell viability of each group was detected by CCK8 method. The expression of Bmal1, Per1, Wee1, Cdk1, Ccnb1 and Mmp13 mRNA in each group was detected by RT-qPCR. The expression of BMAL1, PER1, WEE1, CDK1, CCNB1 and MMP13 protein in each group was detected by Western blot. The effects of Bmal1 on different stages of cell cycle and apoptosis was analyzed by flow cytometry. The regulation of Bmal1 on Per1, Wee1, Cdk1, Ccnb1 and Mmp13 and their roles in OA were analyzed. Results Compared with the normal group, the cell viability of the OA group was increased, the relative mRNA expression of Bmal1 and Wee1 in the OA group decreased, and the relative mRNA expression of Per1, Cdk1, Ccnb1 and Mmp13 increased significantly. The cell viability of LV-Bmal1 group decreased, the relative expression of Bmal1 and Wee1 mRNA increased, and the relative expression of Per1, Cdk1, Ccnb1 and Mmp13 mRNA decreased(P＜0.05). Correlation analysis showed that Bmal1 was positively correlated with Wee1 and they were negatively correlated with Per1, Cdk1, Ccnb1 or Mmp13. The results of Western blot showed that protein expression in different groups were consistent with the trend of PCR. The results of cell cycle and apoptosis showed that compared with the normal group, the S phase and G2/M phase of the OA group were shortened, the proportion of cells decreased significantly, and the proportion of early and late apoptosis increased. The S phase and G2/M phase of the LV-Bmal1 group were prolonged, the proportion of cells was increased, and the proportion of early and late apoptosis was decreased. Conclusions Circadian clock gene Bmal1 in inflammatory chondrocytes might regulate the expression of cell cycle-related genes. [ABSTRACT FROM AUTHOR]

To explore the effect of sorafenib on HeLa cell proliferation by inducing cell apoptosis and autophagy and its impact on drug resistance. Methods The drug-resistant cell strains were constructed through intermittent induction method, with concentrations of 0, 2.5, 5.0, 7.5, 10.0, 15.0, 20.0 μmol/L. HeLa cells were incubated with increasing concentrations of sorafenib with each concentration for 1 week. The drug-resistant cell strains with stable passages were collected. MTT assay was used to detect the effect of sorafenib on cell proliferation. Cell cycle distribution was analyzed by flow cytometry. The change in the expression of drug-resistant and apoptotic genes in the parents and drug-resistant cell strains under different drug concentrations was examined by semi-quantitative PCR. The changes of apoptotic related marker proteins LC3-Ⅰ and LC3-Ⅱ were detected by Westernblot. Results Stable drug-resistant strains were successfully obtained; Drug-treated cells were more blocked in the G1 phase. In drug-resistant cells, the expression of apoptosis suppressor gene Bcl-2 was significantly decreased and the apoptotic gene Bax as well as the drug-resistant genes were all significantly increased(P＜0.05). The LC3-Ⅱ/LC3-Ⅰ ratio of drug-resistant cells was significantly higher than that of parent cells(P＜0.05). Conclusions Sorafenib may block the cell cycle, suppress malignant cell proliferation and promote autophage. On one hand, autophagy participates in the development of cell drug resistance and promotes cell survival. On the other hand, drug-induced autophagy may activate some of apoptotic signaling pathway in drug-resistant cells and promote the reversal of cell drug resistance. [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.)

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.)

Copyright of Journal of South China University of Technology (Natural Science Edition) is the property of South China University of Technology 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.)

To investigate the effects of low expression of microRNA21 (miR21) on the proliferation and apoptosis of pituitary tumor cell line RC4BC, and to analyze its targeting relationship with phosphatase and tensin homolog deleted on chromosome ten (PTEN) . Methods RC4BC cells in the logarithmic growth phase were divided into two groups. Cells in the silence group were transfected with miR21 inhibitor, while cells in the negative control group were transfected with inhibitor negative control NCinhibitor. RTPCR was used to detect the mRNA of miR21 and PTEN. CCK8 assay was used to observe the cell proliferation ability of the two groups (expressed by OD value）, plate cloning assay was used to observe the cell colony formation ability of the two groups (expressed by colony formation number）, flow cytome⁃ try was used to observe the apoptosis rates of the two groups and to observe the cell cycle distribution. Single cell suspension was prepared from RC4BC cells, and miR21 mimics or NCmimics were cotransfected into RC4BC cells with PTENWT or PTENMUT, respectively. After transfection, the cells were labeled as the miR21 mimics+PTENWT group, NCmim⁃ ics+PTENWT group, miR21 mimics+PTENMUT group, and NCmimics+PTENMUT group, respectively. The dual lu⁃ ciferase reporter gene assay was used to verify the targeting relationship between miR21 and PTEN. Results The relative expression levels of miR21 and PTEN mRNA in RC4BC cells of the silence group were 0. 30±0. 08 and 2. 89±0. 14, re⁃ spectively, while those in the negative control group were 1. 01±0. 02 and 0. 99±0. 03, respectively, with statistically sig⁃ nificant differences (all P<0. 05) . The OD values of RC4BC cells of the silence group were lower than those of the nega⁃ tive control group at 24，48, and 72 h (all P<0. 05) . The number of colony formation of RC4BC cells in the silence group was smaller than that in the negative control group (P<0. 05) . The apoptosis rate of RC4BC cells in the silence group was higher than that in the negative control group (P<0. 05) . The proportions of cells in the G0/G1 phase and S phase were 65. 65%±7. 82% and 19. 25%±3. 70%, in the silence group, respectively, versus 45. 62%±5. 03% and 35. 72%±4. 67%, in the negative control group, respectively, with statistically significant differences (all P<0. 05) . The relative luciferase activity of the miR21 mimics+PTENWT group, NCmimics+PTENWT group, miR21 mimics+PTENMUT group, and NCmimics+PTENMUT group was 0. 39±0. 07，1. 02±0. 03，1. 01±0. 04, and 1. 00±0. 03, respectively, and significant difference was found in the relative luciferase activity between the miR21 mimics+PTENWT group and the other groups (all P<0. 05) . Conclusion Silencing miR21 can inhibit the proliferation and promote apoptosis of RC4BC cells in pi⁃ tuitary tumor cell line, and its mechanism may be related to the targeted regulation of PTEN gene. [ABSTRACT FROM AUTHOR]