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Background: Oral squamous cell carcinoma (OSCC) refers to the malignant tumor of the head and neck with a highest morbidity. It exhibits a poor prognosis and unsatisfactory treatment, which is partially attributed to delayed diagnosis. As indicated from existing reports, the protein histidine phosphatase LHPP acts as a vital factor in tumorigenesis in liver, lung, bladder, breast and pancreatic tumor tissues. Thus far, the expression level of LHPP in OSCC has been rarely studied, and its functional mechanism remains unclear. Methods: DEG analysis, OSCC cell lines and OSCC samples were used to detect the expression of LHPP in adjacent normal and cancerous tissues and its relationship with OSCC differentiation. Immunofluorescence staining was used to detect the over-expressed LHPP in OSCC cell lines. The cell counting kit 8 test, EdU proliferation test, scratch test, invasion test, single clone formation test, mouse xenograft tumor model HE staining and immunohistochemistry were used to estimate the biological characteristics of LHPP in OSCC in vivo and in vitro. GO and KEGG enrichment analysis and LHPP transcription factor analysis were used to further predict the role of LHPP and its related genes. The Western blotting assay, real-time PCR analysis and flow cytometry determined the inhibitory mechanism of LHPP in OSCC cells.Results: LHPP was down-regulated in OSCC tissues and cells than that in normal oral mucosa tissues and cells, LHPP expressing also displays a close association with the degree differentiation of OSCC. LHPP is capable of significantly inhibiting OSCC cells from proliferating, migrating and invading. The increase in LHPP expression facilitated OSCC cell apoptosis through PI3K/AKT signaling pathway.Conclusion: LHPP is a novel tumor suppressor which promotes apoptosis by inhibiting the PI3K/AKT signaling pathway in OSCC, indicating that LHPP is a new diagnostic marker and therapeutic target for OSCC.

18β‐Glycyrrhetinic acid (18β‐GA) is a component extracted from licorice. This study aimed to evaluate the effects of 18β‐GA on isoproterenol (ISO)‐induced acute myocardial infarction in rats and mice. Two consecutive days of subcutaneous injection of ISO (85 mg/kg/day) resulted in acute myocardial infarction. We examined the pathological changes, oxidative stress, inflammatory response, and expression of apoptosis in mouse hearts. The expressions of phosphoinositol‐3‐kinase (PI3K), protein kinase B (Akt), and the phosphorylation levels of PI3K (p‐PI3K) and Akt (p‐Akt) were determined by western blotting. The whole‐cell patch‐clamp technique was applied to observe the L‐type Ca2+ currents, and the Ion Optix detection system was used for cell contraction and Ca2+ transient in isolated rat cardiac ventricular myocytes. In ISO‐induced myocardial infarction, the J‐point, heart rate, creatine kinase, lactate dehydrogenase, superoxide dismutase, catalase, malondialdehyde, glutathion, and reactive oxygen species decreased in mice after 18β‐GA treatment. 18β‐GA improved ISO‐induced morphologic pathology, inhibited the inflammatory pathway response and cardiomyocyte apoptosis, and inhibited PI3K/Akt signaling. 18β‐GA could significantly inhibit ICa‐L, myocardial contraction, and Ca2+ transient. This study demonstrates that 18β‐GA has cardioprotective effects on acute myocardial infarction, which may be related to inhibiting oxidative stress, inflammation, apoptosis via the PI3K/Akt pathway, and reducing cell contractility and Ca2+ concentration via L‐type Ca2+ channels., MI has become one of the most common cardiovascular diseases. Because of the difficulty of clinical treatment, preventing and improving MI is meaningful and challenging work. Therefore, it is very necessary to identify drugs that are effective against MI. Our study demonstrates that 18β‐GA has cardioprotective effects on acute MI. 18β‐GA can inhibit oxidative stress, inflammatory response, apoptosis through the PI3K/Akt signaling pathway and can also reduce cell contractility and Ca2+ influx through LTCC.

Hepatocellular carcinoma (HCC) is a common type of primary liver cancer. Circular RNAs (circRNAs) have been demonstrated to be a crucial player in multiple cancers. However, a large number of circRNAs remain to be explored. Our study focused on investigating hsa_circ_0004018 in HCC. Firstly, we conducted quantitative reverse transcription PCR (RT-qPCR) to find that circ_0004018 was down-regulated in HCC cells. Western blot analysis was performed to detect the protein levels of phosphatase and tensin homologue (PTEN) and related factors of PI3K/AKT signaling pathway. From the results of functional assays, we found that overexpression of circ_0004018 significantly inhibited the proliferative and migratory capacities of HCC cells. The regulatory mechanism of circ_0004018 in HCC was determined by RNA immunoprecipitation (RIP), RNA pull-down, and luciferase reporter assays, thereby we knew that circ_0004018 regulated PTEN by sequestering microRNA-1197 (miR-1197) to modulate PI3K/AKT signaling pathway. Finally, rescue assays verified that circ_0004018 participated in modulation of cell proliferation and migration in HCC via sponging miR-1197 and regulating PTEN. In conclusion, circ_0004018 suppresses the proliferation and migration of HCC cells via sponging miR-1197 to inactivate the PTEN/PI3K/AKT signaling pathway. Abbreviations: HCC: Hepatocellular carcinoma; circRNAs: Circular RNAs; PTEN: Phosphatase and tensin homologue; miR-1197: microRNA-1197; ceRNA: competitive endogenous RNA; ATCC: American Type Culture Collection; EMEM: Eagle���s Minimum Essential Medium; RT-qPCR: Quantitative real-time PCR; EdU: 5-ethynyl-20-deoxyuridine; FISH: Fluorescent in situ hybridization; RIP: RNA immunoprecipitation; 3��-UTR: 3��-untranslated region; Wt: wild-type; Mut; mutant type; gDNA: genomic DNA; Act D: Actinomycin D; PI3K: phosphatidylinositol-3-kinase; AKT: protein kinase; lncRNAs: long non-coding RNAs

Spinal cord injury (SCI) is a traumatic injury leading to hypoxia, lipid peroxidation, and cellular apoptosis. Honeysuckle (Lonicera japonicae) has been used to treat headaches and play a neuroprotective role. However, its possible effects on protection from neurological function loss during injury remain to be explored. To this end, we have investigated the effects of honeysuckle extract on neural function injury in SCI rats and report an improvement in nerve function. The mechanism involved reduced expression of proinflammatory cytokines, decrease in cellular apoptosis, and activation of PI3K/Akt pathway. We conclude that possible use of honeysuckle extract for treatment of SCI needs further exploration.

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive fibrotic interstitial pneumonia with unknown causes. The incidence rate increases year by year and the prognosis is poor without cure. Recently, phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB/AKT) signaling pathway can be considered as a master regulator for IPF. The contribution of the PI3K/AKT in fibrotic processes is increasingly prominent, with PI3K/AKT inhibitors currently under clinical evaluation in IPF. Therefore, PI3K/AKT represents a critical signaling node during fibrogenesis with potential implications for the development of novel anti-fibrotic strategies. This review epitomizes the progress that is being made in understanding the complex interpretation of the cause of IPF, and demonstrates that PI3K/AKT can directly participate to the greatest extent in the formation of IPF or cooperate with other pathways to promote the development of fibrosis. We further summarize promising PI3K/AKT inhibitors with IPF treatment benefits, including inhibitors in clinical trials and pre-clinical studies and natural products, and discuss how these inhibitors mitigate fibrotic progression to explore possible potential agents, which will help to develop effective treatment strategies for IPF in the near future., Graphical abstract PI3K/AKT signaling participates in the formation of idiopathic pulmonary fibrosis (IPF), and some promising PI3K/AKT inhibitors already show IPF treatment benefits, making targeting PI3K/AKT a novel anti-fibrotic strategy.Image 1

Chemo-resistance and metastasis are the most common causes of breast cancer recurrence and death. Thidiazuron (TDZ) is a plant growth regulator (phytohormone) whose biological effects on humans and animals has not yet been determined. In this study, we investigated the anticancer activity of this phytohormone on the drug resistant-triple negative breast cancer cell line MDA-MB-231. Treatment of the breast cancer cells with TDZ (1–50 μmol/L) caused more stressful environment and induced a significant increase in active caspase-positive cells. In addition, TDZ treatment (5 and 10 μmol/L) significantly attenuated the migration and the invasiveness of these highly metastatic cancer cells. Mechanistically, TDZ reduces cancer progression and invasiveness by targeting miR-202-5p, which stimulates the expression of phosphatase and tensin homolog (PTEN), the tumor suppressor that downregulates the PI3K–Akt signaling pathway. Treatment with TDZ significantly upregulates miRNA-132, the suppressor of breast cancer proliferation, which is also implicated in dysregulation of the TEN-Akt–NFκB signaling pathway. Interestingly, our molecular docking analysis revealed a potential non-covalent interaction between TDZ and Akt, PTEN, and PI3K. These findings suggest that TDZ suppresses breast cancer metastasis by targeting miRNA-132, the miR-202-5p–PTEN axis, and the PI3K–Akt signaling pathway downstream.

AIM: To investigate whether the curcumin reduce retinal inflammation in animal model and human retinal pigment epithelium (ARPE)-19 cells. METHODS: In vivo, male C57/B6 mice received intraperitoneal injections of curcumin for 3d before intraperitoneal injection of lipopolysaccharide (LPS; 10 mg/kg) to induce retinal inflammation. 24h after LPS application, the mRNA levels of pro-inflammatory cytokines were detected by real-time polymerase chain reaction (RT-PCR). Concanavalin A lectin perfusion-labeling technique evaluated leukocyte adhesion to the retinal vasculature. The protein concentration in the anterior chamber was measured with a protein quantification kit. In vitro, ARPE-19 cells were cultured. The optimum concentration of curcumin was detected by cell counting kit-8 (CCK-8) assay. Before stimulated with 5 μg/mL LPS, ARPE-19 cells were incubated with or without curcumin for 1h. Pro-inflammatory cytokines were measured by RT-PCR and ELISA. PI3K/Akt expression was analyzed by Western blotting. RESULTS: Curcumin pre-treatment led to significant inhibition of EIU-associated leukocyte adhesion to retinal blood vessels and anterior-chamber protein leakage. The mRNA expression level of inflammatory cytokines was also significantly reduced with application of curcumin in vivo, such as IL-1β, IL-6 and TNF-α. Meanwhile, Curcumin significantly attenuated the expression of IL-6, IL-8 and MCP-1 at both mRNA and protein levels in ARPE-19 cells. Curcumin suppressed PI3K/Akt phosphorylation as well as NF-κB activation in LPS-activated ARPE-19 cells. CONCLUSION: Curcumin plays a preventive effect on LPS-induced retinal inflammation. The beneficial effect appears associated with inhibiting of the PI3k/Akt signaling pathway.

Breast cancer (BCa) is one of the most common gynecological malignancies. Ubiquitin-coupled enzyme E2T (UBE2T) has been demonstrated to play crucial roles in various tumors.UBE2T levels were detected using quantitative real time PCR and western blot. CCK-8 and colony formation assays were used to evaluate cell proliferation. A xenograft model was used to evaluate the effects of UBE2T on tumor growth in mice, and immunohistochemistry (IHC) assay was performed to detect the expression of UBE2T and Ki-67. Transwell assay was performed to determine cell migration and invasion. The ATP level, glucose consumption and lactate production were measured using the corresponding commercial kits. Western blot assay was used to detect the levels of epithelial-mesenchymal transformation (EMT), glycolytic and the PI3K/AKT pathway related proteins regulated by UBE2T.Upregulation of UBE2T expression in human BCa tissues was found in human clinical BCa tissues and The Cancer Genome Atlas (TCGA) dataset. The expression of UBE2T was confirmed to be up-regulated in BCa cells compared to normal breast epithelial cell line (MCF-10A). Overexpression of UBE2T promoted proliferation, migration, invasion and glycolysis in BCa cells, while UBE2T knockdown showed the opposite results. Moreover, UBE2T knockdown suppressed tumor growth in mice. Further mechanism analysis shows that UBE2T participated in the regulation of BCa progression through affecting the PI3K/AKT signaling pathway.UBE2T promoted proliferation, invasion and glycolysis through modulating PI3K/AKT signaling pathway in BCa, implying that UBE2T may provide a promising therapeutic target for the therapy of BCa.

Herein, we purposed to explore whether hypoxia triggers proliferation of cholesteatoma keratinocytes via the PI3K-Akt signaling cascade. Cells were inoculated with different concentration of CoCl2. The proliferation and cellular HIF-1α, p-PDK1 and p‑Akt expression levels of cholesteatoma keratinocytes were assessed in vitro. Hypoxia escalated cell proliferation via upregulating p-PDK1 and p‑Akt expressions. Specific inhibitor of the PI3K-Akt signaling cascade, LY294002 markedly inhibited the expression of p‑Akt and significantly reduces the hypoxia‑induced proliferation of cholesteatoma keratinocytes. Our data provides research evidence confirming that hypoxia participates in the onset and progress of cholesteatoma.

Cancer Biotherapy and Radiopharmaceuticals is officially retracting the article entitled, A Novel CircRNA Circ_0095424 Regulates Proliferation, Metastasis, and Apoptosis of Osteosarcoma Cells Via the PI3K/AKT Signaling Pathway Through Targeting the miR-1238/HMGB1 Axis by Zhang et al., (Cancer Biother Radiopharm epub 19 Aug 2020; DOI: 10.1089/cbr.2020.3563), due to manipulated images appearing in the published paper. The Editor of the journal received an email on August 31, 2020 from the corresponding author of the article, Dr. Chuan Qin, indicating that, ''due to our negligence in organizing the pictures, the protein pictures are repeatedly placed in Figure 7G PI3K. For this, we express our sincerest apologies. We need to [issue] an [erratum] on this issue. We have replaced the protein picture of Figure 7G with the correct picture.'' However, one of the attachments submitted with the request appeared to be the original version of Figure 7 from the accepted manuscript. A second attachment appeared to be the data from three replicates to be used (by the journal) to construct a revised version of Figure 7. The Editor, in turn, informed the authors that it was not at the journal's discretion to create a new image for them, and asked the authors to create the revised figure and supply it to the publisher. Below is the response from Dr. Qin, dated September 2, 2020. "In fact, our team's Western blot experiment commissioned a third-party company for testing. At present, some peers have found that the company has forged experimental reports. We believe that the authenticity of the data provided by the company is problematic. After contacting the company, they were unable to provide the original images. In view of the problems in this manuscript, all the authors discussed and agreed to withdraw the manuscript." As the entirety of the situation is unacceptable, the Editor officially retracts the article based on the "forged experimental reports" and the questionable validity of the data provided. The Editor and Publisher of Cancer Biotherapy and Radiopharmaceuticals is dedicated to preserving the integrity of the scientific literature and the community it serves.

Decursin, a coumarin compound isolated from Angelica gigas has been shown to possess multiple anti-tumor activities. But it’s still little known about the effects associated with cervical cancer. T...

Decreased expression of the long non-coding RNA HOXD-AS2 promotes gastric cancer progression by targeting HOXD8 and activating PI3K/Akt signaling pathway

Objective: To assess sorafenib's role in regulating DJ-1-PTEN/PI3 K/AKT pathway and affecting the biological effects of liver cancer cells. Methods: Human normal liver HL-7702, liver cancer MHC97-L, and HCCLM3 cells were cultured in vitro to measure DJ-1 and PTEN expression. MHC97-L and HCCLM3 cells were treated with 0, 1.0, 2.0 M of sorafenib followed by analysis of cell viability by CCK-8, and DJ-1 and PTEN expressions by Western blot. MHC97-L cells were separated into control group, 2.0 M Sorafenib treatment group, Sorafenib + pcDNA3.1-Blank group, and Sorafenib + pcDNA3.1-DJ-1 group. Cell proliferation was assessed by flow cytometry and EdU staining. Results: Compared with HL-7702 cells, DJ-1 expression was significantly increased, while PTEN level was significantly declined in MHC97-L and HCCLM3 cells. Sorafenib treatment significantly inhibited the proliferation activity of MHC97-L and HCCLM3 cells. Different concentrations of sorafenib significantly downregulated DJ-1 expression and enhanced PTEN expression in MHC97-L cells. pcDNA3.1-DJ-1 transfection on the basis of sorafenib treatment significantly elevated DJ-1 and p -AKT levels, reduced PTEN expression, decreased cell apoptosis, and enhanced cell proliferation. Conclusion: Sorafenib downregulates DJ-1 expression and affects PTEN/PI3K/AKT pathway activity to exert an anti-tumor effect that inhibits liver cancer cell proliferation and promotes cell apoptosis.

Background Nasopharyngeal carcinoma (NPC) is a common malignant tumor in china. T63 is one of novel curcumin analogues, which reduces the tumorigenic potential effectively. Methods We investigated the mechanism of anti-cancer activity on NPC cells treatment with T63 for the first time. Cell viability was monitored using the method of MTT and colony formation assay. Morphological changes observed by fluorescence microscope with Hoechst33342 staining. The cell cycle, the rates of cell apoptotic and mitochondrial membrane potential were identified by flow cytometry, and apoptotic proteins were identified by western blot analysis. Results In our study, we found a significant decrease in the number of viable cells and in colony formation abilities of NPC cells (CNE2 and CNE2R) after 24, 48 and 72 h treatment with T63. T63 promoted morphologic changes of apoptosis and increased apoptotic rate. T63 promoted also cell cycle arrest of G2/M phases followed by decreased cell viability in CNE2 and CNE2R after 48 h. Then the loss of the mitochondrial membrane potential and release of cytochrome-c were demonstrated. Western blotting analysis found a decrease in Bcl-2/Bax ratio and activation of caspase-8, -9, -3 and PARP. And the changes of PTEN and p-Aktp473 suggested PTEN/PI3K/Akt might be an important signaling pathway in the anticancer role of T63. Conclusions T63 definitely induced apoptosis in radiosensitive and radioresistant NPC cell lines. But the apoptotic effect was stronger on radioresistant cells than that on radiosensitive cells through activation of mitochondria-mediated apoptotic pathway.

This study aimed to explore the role of miR-146b-3p in acute respiratory distress syndrome in septic mice. Ten mice were randomly selected as normal group (n = 10, without any treatment) and 60 septic mice with acute respiratory distress syndrome were divided into model group (n = 10, without any treatment), negative control (NC) mimic group (n = 10, injected with NC mimic), miR-146b-3p mimic group (n = 10, injected with miR-146b-3p mimic), si-NC group (n = 10, injected with PI3Kγ siRNA NC), si-PI3Kγ group (n = 10, injected with PI3Kγ silencing plasmid), and miR-146b-3p mimic + oe-PI3Kγ group (n = 10, injected with miR-146b-3p mimic + PI3Kγ overexpression plasmid). We found that miR-146b-3p negatively regulated PI3Kγ. Compared with normal group, model mice had decreased expression of miR-146b-3p, increased expressions of PI3Kγ, p-AKT, ASC, NLRP3 and Caspase-1 proteins, higher W/D ratio, and more serum IL-1β and IL-18 content (all P

Cervical cancer ranks the second in the incidence of common women malignant tumors, which is the commonest malignant tumor in women's reproductive organs. In present research, we evaluated the possible antitumor effect of miR-181 on cervical cancer and explored the molecular mechanism of this phenomenon. Briefly, MTT, LDH activity kit and flow cytometry were used to detect cell growth, LDH activity and cell apoptosis rate of HeLa cells. The experiment correlation protein expression levels of PTEN, phosphatidylinositol-3-hydroxykinase (PI3K) and phosphorylation (p)-Akt (protein kinase B) were measured by Western blot analysis. In present research, this research observed that miR-181 weakened the cell proliferation activity of cervical cancer cells and induced their caspase3/9 to promotion of cell apoptosis. PTEN expression was induced at the genetic and protein levels, and the PI3K/Akt expressions were inhibited at the protein levels in cervical cancer cells. SiPTEN weakened the antitumor effect of miR-181 on cervical cancer by activation of PI3K/Akt signaling pathway.

Objective: To assess promethazine's effect on myocardial cells in rats with myocardial ischemiareperfusion injury (MIRI). Methods: The rat MIRI model was established and treated as the ischemia group. MIRI rats were treated with promethazine and included as the drug group. Rats only undergoing thoracotomy were enrolled as the control group. The physiological function of heart was assessed using the ultrasound cardiotachograph, and the apoptosis and proliferation of myocardial cells were detected using TUNEL assay and Ki67 staining, respectively. Moreover, the expressions of Caspase-3, Bcl-2, PI3K, GSK-3, PDK-1 and PKB were determined via Western blotting and qPCR. Results: There were significant differences in cardiac function indexes [left ventricular enddiastolic diameter (LVEDd), left ventricular end systolic diameter (LVESd), ejection fraction (EF) and fractional shortening (FS)] among the three groups (p= 0 002, 0.004, 0.025 and 0.012), and ischemia group had the highest LVEDd [(8.73± 0.31) mm] and LVESd [(7.98± 0.37) mm] and lowest EF [(42± 3.8)%] and FS [(40.3± 2.8)%]. The number of apoptotic myocardial cells was significant higher in ischemia group than control ( p< 0 05), while it was significantly declined after treatment with promethazine ( p< 0 05). Caspase-3 was significantly upregulated and Bcl-2 was downregulated in ischemia group which were all significantly reversed in drug group. Besides, Ki67 level was significantly reduced in ischemia group compared to control and higher in drug group than ischemia group, indicating that drug treatment increased cell proliferation ability. The levels of PI3K, GSK-3 and PKB in myocardial tissues were significantly declined in ischemia group and elevated after the treatment with promethazine without difference of PDK-1 level in myocardial tissues among the three groups. Conclusion: Promethazine inhibits apoptosis and promotes proliferation of myocardial cells in MIRI rats through PI3K/Akt signaling pathway.

It is acknowledged that hepatocellular carcinoma (HCC) cause the most cancer-associated mortality all around the world and the traditional Chinese medicine has been widely used in HCC treatment. Physalin B (PB) is the most important chemical compounds of a Chinese traditional herb, Physalis alkekengi var. franchetii. In this research, it is the first time in the world that we illustrated PB exhibits obvious apoptotic activity in SMMC-7721 and HepG2 human HCC cell lines in the does-dependent way. Moreover, decrease in mitochondrial membrane potential (MMP) was found and the cellular reactive oxygen species (ROS) was obviously upregulated in PB-treated cell lines. Furthermore, the expression of apoptosis-associated proteins and ROS were increased after the treatment of PB, which were reversed by the inhibitor of TGF-β, NF-κB or PI3K/AKT signaling pathways. Therefore, PB illustrates potent anticancer activity, which might be studied as a potentiation therapeutic medicine for HCC.

Objective: Studies have shown that Rehmanniae Decoction (Reh) has therapeutic effect on vascular dementia (VD). PI3K/AKT signaling regulates oxidative stress damage and cell apoptosis. Our study intends torehmanniae decoction's effect on the neural function in VD mice. Methods: The mice were divided into Sham group, VD group, low dose Reh+ VD group and high dose Reh+ VD group. Water maze test was used to assess learning and memory ability. The activity of caspase-3, the content of MDA and the activity of SOD enzyme in hippocampus were detected. In vitro , HT22 cells were divided into control group, I–R group, I–R+ 2% Reh serum, I–R+ 4% Reh serum. Flow cytometry was used to detect the intracellular content of ROS and cell apoptosis. Results: Compared with sham group, the learning and memory ability of mice in VD group was significantly decreased. p-AKT level and SOD activity in the hippocampus was decreased, the Caspase-3 activity and MDA content was significantly increased. After treatment of Reh, the learning and memory ability of VD model mice was significantly improved, p-AKT protein expression and SOD activity were up-regulated, and Caspase-3 activity and MDA content were reduced. Conclusion: Rehmanniae decoction alleviates the oxidative stress and inhibits cell apoptosis to improve the function of brain by regulating PI3K/AKT pathway.

Objective: The most commonly reported primary brain tumor in adults is glioma. Choline kinase alpha (CHKA) has been proved to play important roles in glioma. However, the mechanism of CHKA involved...

Background: Colon adenocarcinoma is one of the most common gastrointestinal malignancies with poor prognosis and high mortality. The mRNA-miRNA-lncRNA regulatory network mediated by m6A methylation plays an important role in a variety of cancers including colon adenocarcinoma. Methods: We integrated and analyzed the gene expression data and clinical information of 473 patients with colon adenocarcinoma and 41 normal samples in The Cancer Genome Atlas database. The luciferase reporter gene experiment is used to detect the targeting effect between gene, miRNA and lncRNA. Real-time PCR and Proliferation assays were performed to detect the biological function of gene, miRNA and lncRNA. Results: A risk model and Nomogram that could accurately predict the survival time of patients were constructed through informatics analysis. HYOU1, AL161729.4, miR-760 are differentially expressed in COAD patients and normal samples and are significantly related to survival, and there is a targeted binding effect between the three. Conclusions: HYOU1-AL161729.4-miR-760 ceRNA regulatory network could regulate the proliferation of SW620 cells through the PI3K/Akt signaling pathway.

Background: Zearalenone (ZEA) is widely derived from moldy cereal grain, which has adverse effects on animal reproduction. In particular, pigs are more sensitive to ZEA-induced toxicity than other animals. Isorhamnetin, a flavonoid has extensive of pharmacological activities. However, little is known about the effects of isorhamnetin on reproduction. Thus, it would be interesting to clarify the effect and the underlying molecular mechanism of isorhamnetin involvement in ZEA-induced cytotoxicity in porcine granulosa cells.Results: Our findings showed that isorhamnetin suppressed ZEA-induced apoptosis by regulating Bcl-2 and Bax protein changes. Changes in intracellular Ca2+ levels and CHOP, ATF6, GRP78 indicated that isorhamnetin rescued ZEA-induced endoplasmic reticulum stress (ERS). Furthermore, isorhamnetin prevented ZEA-induced reactive oxygen species (ROS) via P38 signaling pathway. Mechanistically, isorhamnetin stimulated the expression of PCNA and CyclinD, thereby raising the ratio of S phases cells in response to ZEA-induced apoptosis via PI3K/Akt signaling pathway. Isorhamnetin also recovered ZEA-induced steroidogenesis disorder by regulating steroidogenic enzyme gene and proteins (FSH-R, CYP19A1). Conclusions: Collectively, these findings show that isorhamnetin protects granulosa cells from ZEA-induced damage via the PI3K/Akt signaling pathway, which promotes proliferation, alleviates steroidogenesis disorder, ERS and oxidative stress.

Lepidium meyenii (Maca) is an annual or biennial herb from South America that is a member of the genus Lepidium L. in the family Cruciferae. This herb has antioxidant, anti-apoptotic, and enhances autophagy functions and can prevent cell death, and protect neurons from ischemic damage. Macamide B, an effective active ingredient of maca, has a neuroprotective role in neonatal hypoxic-ischemic brain damage (HIBD), and the underlying mechanism of its neuroprotective effect is not yet known. The purpose of this study is to explore the impact of macamide B on HIBD-induced autophagy and apoptosis and its potential mechanism for neuroprotection. The modified Rice-Vannucci method was used to induce HIBD on 7-day-old (P7) macamide B and vehicle-pretreated pups. TTC staining was used to evaluate the cerebral infarct volume of pups, brain water content was measured to evaluate the neurological function of pups, neurobehavioral testing was used to assess functional recovery after HIBD, TUNEL and FJC staining was used to detect cell autophagy and apoptosis, and western blot analysis was used to detect the expression levels of the pro-survival signaling pathway phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and autophagy and the apoptosis-related proteins. The results show that macamide B pretreatment can significantly decrease brain damage, improve the recovery of neural function after HIBD. At the same time, macamide B pretreatment can induce the activation of PI3K/AKT signaling pathway after HIBD, enhance autophagy, and reduce hypoxic-ischemic (HI)-induced apoptosis. In addition, 3-methyladenine (3-MA), an inhibitor of PI3K/AKT signaling pathway, significantly inhibits the increase in autophagy levels, aggravates HI-induced apoptosis, and reverses the neuroprotective effect of macamide B on HIBD. Our data indicate that macamide B pretreatment might regulate autophagy through PI3K/AKT signaling pathway, thereby reducing HIBD-induced apoptosis and exerting neuroprotective effects on neonatal HIBD. Macamide B may become a new drug for the prevention and treatment of HIBD.

Background Growing evidence has indicated that tumor-associated macrophages (TAMs) promote tumor angiogenesis. However, the mechanisms underlying the pro-angiogenic switch of TAMs remains unclear. Here, we examined how exosomal miR-301a-3p secreted by esophageal squamous cell carcinoma (ESCC) cells triggers the pro-angiogenic switch of TAMs. Methods We quantified miR-301a-3p levels in ESCC tumors using qRT-PCR. Macrophage phenotypes were identified using flow cytometry and qRT-PCR. The pro-angiogenic ability of TAMs was measured using the CCK-8 assay, scratch assay, Transwell migration and invasion assay, and tube formation assay. The mechanism by which exosomal miR-301a-3p secreted by ESCC cells triggers the pro-angiogenic switch of TAMs was elucidated using western blots, qRT-PCR, and a dual-luciferase reporter assay. Results We observed anomalous miR-301a-3p overexpression in ESCC tumor tissues and cell lines. Then, we verified that ESCC-derived exosomes promoted angiogenesis by inducing macrophage polarization into M2 type, and exosomal miR-301a-3p secreted by ESCC cells was responsible for this effect. Finally, we discovered that exosomal miR-301a-3p promoted M2 macrophage polarization via the inhibition of PTEN and activation of the PI3K/AKT signaling pathway, subsequently promoting angiogenesis via the secretion of VEGFA and MMP9. Conclusion The pro-angiogenic switch of TAMs is triggered by exosomal miR-301a-3p secreted from ESCC cells via the PTEN/PI3K/AKT signaling pathway. Although tumor angiogenesis can be regulated by a wide range of factors, exosomal miR-301a-3p could hold promise as a novel anti-angiogenesis target for ESCC treatment.

Background Laryngeal squamous cell carcinoma (LSCC) is one of the most commonly seen malignancies of the head and neck, with increasing incidence and mortality. The Y-box-binding protein 1 (YBX1) is a type of oncoprotein which is related to the malignant phenotype of many cancers. It is reported that YBX1 could regulate tumorigenesis, recurrence, and metastasis in multiple cancers. However, little is known about its carcinogenic function and mechanism in LSCC. Methods Firstly, Through Oncomine StarBase, we found that the YBX1 mRNA level was increased in a variety of cancer tissues, including in the LSCC, compared with normal tissues. We silenced YBX1 in LSCC cells using short hairpin RNAs (shRNAs). Secondly, the biological function of YBX1 in LSCC cells was examined by the Cell Counting Kit-8 (CCK-8) assay, flow cytometry, the wound healing assay, and the transwell assay. Thirdly, the correlation between YBX1 and the PI3K/AKT pathway was verified by the western blot assay. Results Expression of YBX1 is higher in a variety of cancer tissues, especially in the head and neck cancers. After transfected with lentiviral vectors, the expression of YBX1 was significantly silenced. Functionally, low expression of YBX1 promoted LSCC cell apoptosis and inhibited LSCC cell proliferation, migration, and invasion. The transfection of sh-YBX1 resulted in an obvious decrease in PI3K/AKT signaling molecules in LSCC cells. Conclusions We demonstrated that YBX1 could promote LSCC cell progression through the PI3K/AKT pathway, providing new insights into a potential biomarker and target for LSCC treatment.

Background: Generally, both strands of a single pre-miRNA have been demonstrated to play a similar role in the same tumor type. However, there are no available literatures yet so far clarifying the opposite roles of both strands froma single miRNAin one tumor type. The purpose of this study is to investigate the functional role of both strands of miR-154 in bone metastasis of prostate cancer (PCa). Methods: miR-154-5p expression was examinedin 285 clinicalPCa tissuesby in situ hybridization. The clinical correlation ofmiR-154-5p expression with clinicopathological features,and overall and bone metastasis-free survival inPCa patients was evaluated by Kaplan-Meier survival and statisticalanalysis. The biological roles of miR-154-3p and miR-154-5p in the bone metastasis of PCa were investigated both in vitroand in vivo.Bioinformatics analysis, western blot and luciferase reporter analysis were used to determinethe potential targets of miR-154-5p.Luciferase assay and Western blotting were performed to clarify the underlying pathway implicated in the role of miR-154-5p in bone metastasis of PCa.Results: Contrary to the well established pro-bone metastatic role of miR-154-3p in PCa, we found that miR-154-5p expression was reduced in PCa tissues with bone metastasis andbone metastatic PCa cell lines. Downexpression of miR-154-5p was positively associated with bone metastasis status, and predicted poorer bone metastasis-free survival in PCa patients. Gain of function experiments showed that upregulating miR-154-5p repressed, while silencing miR-154-5p promoted invasion, migration and proliferation capacities of PCa cells in vitro. Conversely, miR-154-3p yielded an opposite effect oninvasion and migration capacities of PCa cells. Importantly, administration of agomir-154-5p effectivelyinhibited bone metastasis of PCa cellsin vivo. Mechanistic dissection further demonstrated miR-154-5p inhibited invasion, migration and proliferation by targeting EGFR and FGFR1, leading to inactivation of PI3K/AKT signaling. However, the autocrine levels of corresponding ligands in the supernantant of PCa cells were not affected by the changed expression of miR-154-5p.Conclusion: Our results for the first time reveal the different role of both strands froma single miRNA in bone metastasis of PCa, which will facilitate the development of anti-bone metastatic therapeutic strategy in PCa.

Background Cholangiocarcinoma (CCA) is one of the deadliest cancers of the digestive tract. The prognosis of CCA is poor and the 5-year survival rate is low. Bioinformatic analysis showed that early mitotic inhibitor 2 (EMI2) was overexpressed in CCA but the underlying mechanism is not known. Methods The data on bile duct carcinoma from TCGA and GEO databases were used to detect the expression of EMI2. The transcription factors of EMI2 were predicted using JASPAR and PROMO databases. Among the predicted transcription factors, YY1 has been rarely reported in cholangiocarcinoma, and was verified using the luciferase reporter gene assay. RT-PCR was performed to predict the downstream pathway of EMI2, and PI3K/Akt was suspected to be associated with it. Subsequently, in vivo and in vitro experiments were conducted to verify the effects of silencing and overexpressing EMI2 and YY1 on the proliferation, invasion, and metastasis of the bile duct cancer cells. Results EMI2 was highly expressed in CCA. Silencing EMI2 inhibited the proliferation, invasion, and migration of CCA cells, arrested cell cycle in the G1 phase, and promoted of apoptosis. The luciferase reporter gene assay showed that YY1 bound to the promoter region of EMI2, and after silencing YY1, the expression of EMI2 decreased and the progression of CCA was inhibited. Moreover, key proteins in the PI3K/Akt signaling pathway decreased after silencing EMI2. Conclusion EMI2 may be one of the direct targets of YY1 and promotes the progression of CCA through the PI3K/Akt signaling pathway.

Melanoma is considered as the most frequent primary malignancy occurring in skin. Accumulating studies have suggested that long non-coding RNAs (lncRNAs) play critical parts in multiple cancers. In this study, we explored the molecular mechanism of ZBED3 antisense RNA 1 (ZBED3-AS1) in melanoma. We observed that ZBED3-AS1 expression was remarkably up-regulated in melanoma tissues, and high ZBED3-AS1 level was linked to unsatisfactory survival of melanoma patients. Then, we discovered that ZBED3-AS1 was overexpressed in melanoma cells compared with human epidermal melanocytes. In addition, loss-of-function assays verified that ZBED3-AS1 knockdown restrained cell proliferation, migration, epithelial-mesenchymal transition (EMT), and stemness in melanoma. In addition, signal transducer and activator of transcription 3 (STAT3), which also showed tumour-facilitating functions in melanoma, was confirmed as a transcriptional activator of ZBED3-AS1. Moreover, ZBED3-AS1 enhanced the expression of AT-rich interaction domain 4B (ARID4B) through sequestering miR-381-3p. Importantly, we further confirmed that ZBED3-AS1 promoted the malignant progression of melanoma by regulating miR-381-3p/ARID4B axis to activate the phosphatidylinositol 3-kinase/AKT serine/threonine kinase (PI3K/AKT) signalling pathway. In a word, our research might provide a novel therapeutic target for melanoma.

Background: Melanoma is a highly aggressive cancer that can metastasize at early stage. The mechanosensitive ion channel Piezo1 plays a crucial role in embryonic development, tumour growth, migration, invasion and vascularization. The aim of this study was to clarify the role of Piezo1 and its potential mechanism in regulating the malignant phenotypes of melanoma.Methods: The expression of Piezo1 in melanoma was analysed using quantitative real-time PCR and public databases. The effect of Piezo1 on cell viability was examined using a cell counting kit-8 assay. Cell invasion and migration ability were assessed using wound healing assays, transwell assays, transendothelial migration assays and a tail vein cancer metastasis model in vivo. Bioinformatics and western blot assayses were used to explore the effect of Pieoz1 on P13K/AKT signalling.Results: Piezo1 was upregulated in melanoma and was positively associated with poor survival. Piezo1 knockdown significantly weakened the intracellular calcium signal significantly and inhibited the viability of melanoma cells. Furthermore, Piezo1 knockdown inhibited the invasion and metastasis ability in vitro and in vivo by inducing the expression of cell cycle, invasion and metastasis related genes. To clarify the possible mechanism, it seems that Piezo1 activates the PI3K-AKT signalling to maintain malignant phenotypes of melanoma.Conclusion: Piezo1 acts as an oncogene in melanoma cells and provides a novel candidate for melanoma diagnosis and treatment.

Background: Cinobufacini injection (CI), an aqueous extraction from the Cutis Bufonis, is broadly used in clinical treatment of cancer in China. However, the underlying molecular mechanisms of CI in treating osteosarcoma (OS) remain unclear. Aberrant activation of PI3K-AKT signaling pathway is the cause of many types of cancer, including OS. Therefore, we investigated the effect of CI on proliferation, apoptosis and cell cycle of OS cells and elucidated the molecular mechanism of CI in inhibiting OS cells. Methods: Cell proliferation of U2OS and MG63 cells after CI treatment was measured by CCK-8 assay, colony formation and morphological changes. Additionally, the cell cycle arrest and apoptosis induced by CI, were determined by FACS and Western blot analysis. The mechanisms of CI on OS were evaluated by RNA-seq and Western blot analysis. Results: We founf that CI reduced the proliferation of U2OS and MG63 cells in a dose- and time- dependent manner. Furthermore, CI induced the U2OS cells cycle arrest in G0/G1 phase, but the MG63 cells cycle arrest in G2/M phase. Consequently, CI triggered the apoptosis in both OS cells, with enhanced caspase-3 activity and decreased expression of Bcl-2/Bax. In addition, RNA-seq data indicated that PI3K-Akt signaling pathway played an essential role in CI treatment. Moreover PI3K and phosphorylation of AKT (p-AKT) were significantly down-regulated by CI in both OS cells. Conclusions: These results indicate that CI significantly inhibited the proliferation, induced the cell cycle arrest, as well as apoptosis in human OS cells, which is mediated by the inactivation of PI3K-Akt signaling pathway. These findings suggest that CI may have potential for the treatment of OS.

Stroke is one of the leading causes of death and disability worldwide with limited therapeutic options. Melatonin can attenuate ischemic brain damage with improved functional outcomes. However, the cellular mechanisms of melatonin-driven neuroprotection against post-stroke neuronal death remain unknown. Here, distal middle cerebral artery occlusion (dMCAO) was performed in C57BL/6j mice to develop an ischemic stroke in vivo model. Melatonin was injected intraperitoneally immediately after ischemia, and 24 and 48 hours later. Melatonin treatment, with 5 to 20 mg/kg, elicited a dose-dependent decrease in infarct volume and concomitant increase in sensorimotor function. At the molecular level, phosphorylation of PTEN and Akt were increased, whereas PTEN activity was decreased in melatonin treated animals 72 hours after dMCAO. At the cellular level, oxygenglucose deprivation (OGD) challenge of neuronal cell line Neuro-2a (N2a) and primary neurons supported melatonin’s direct protection against neuronal cell death. Melatonin treatment reduced LDH release and neuronal apoptosis at various time points, markedly increased Akt phosphorylation in neuronal membrane, but significantly suppressed it in the cytoplasm of post-OGD neurons. Mechanistically, melatonin-induced Akt phosphorylation and neuronal survival was blocked by Wortmannin, a potent PIP3 inhibitor, exposing increased PI3K/Akt activation as a central player in melatonin-driven neuroprotection. Finally, PTEN knock-down through siRNA significantly inhibited PI3K/Akt activation and cell survival following melatonin treatment, suggesting that melatonin protection against ischemic brain damage, is at least partially, dependent on modulation of the PTEN/PI3K/Akt signaling axis.

Context: Fuzhu Jiangtang Granules (FJG) are a traditional Chinese used in the treatment of diabetes mellitus. However, the antidiabetic mechanism of FJG is not clear. Objective: This study evaluates and determines the antidiabetic mechanism of FJG using a Zucker diabetic fatty (ZDF) rat model. Materials and methods: ZDF (fa/fa) rats were divided into four groups (n = 6): diabetes mellitus (DM), metformin (Met, 0.134 g/kg b.w./day), FJG (0.64 g/kg b.w./day), and combination (Com, 0.134 g/kg b.w./day of Met and 0.64 g/kg b.w./day of FJG). Six ZDF (fa/+) rats served as a normal control. After 6 weeks, biochemical parameters gene and protein expression were detected. Results: The FBG, bodyweight, triglyceride (TG), total cholesterol (TC), free fatty acid (FFA), insulin levels, and HOMA-IR were lower in the FJG than in the DM group (p

Certain microRNAs (miRNA/miRs) serve important roles in the progression of atherosclerosis (AS); however, the exact regulatory mechanisms of miRNAs in AS remain to be fully elucidated. The present study aimed to investigate the effects of miR-125a-5p in AS and the underlying mechanisms. Oxidized low-density lipoprotein (ox-LDL) was employed to stimulate human aortic vascular smooth muscle cells (HAVSMCs) to establish a cell model of AS. Reverse transcription-quantitative PCR was used to determine the expression levels of miR-125a-5p. Cell proliferation was evaluated using a Cell Counting Kit-8 (CCK-8) and migration was detected using a Transwell assay. In addition, the levels of the VSMC-specific marker gene α-smooth muscle actin, the cell cycleregulatory proteins cyclin-dependent kinase (CDK)2, cyclin D1, cyclin E and p27, as well as the migration-associated proteins matrix metalloproteinase-2 (MMP2) and MMP9, and phosphorylated phosphoinositide 3-kinase (p-PI3K) and p-AKT were determined by western blot analysis. The results revealed that transfection of miR-125a-5p mimics inhibited the ox-LDL-induced proliferation of HSVSMCs, accompanied by decreased expression of CDK-2, cyclin D1 and cyclin E, and increased expression of p27. Furthermore, miR-125a-5p mimics attenuated the ox-LDL-induced migration of HAVSMCs in parallel with downregulation of the expression of MMP2 and MMP9. Furthermore, the effect of ox-LDL to increase p-PI3K and p-AKT levels was significantly blunted by miR-125a-5p mimics. In conclusion, the present results suggested that miR-125a-5p mimics inhibited ox-LDL-induced proliferation and migration of HAVSMCs through inhibition of PI3K/AKT signaling, providing a potential novel therapeutic strategy for AS.

Oral squamous cell carcinoma (OSCC) is the sixth most prevalent cancer type in the world, with a 5-year survival rate of only 40% to 50%. Finding effective anti-tumor drug candidates is quite necessary for the progression of OSCC therapy. In this study, the role of Thymoquinone (TQ) in OSCC cells was studied. It was confirmed that TQ can inhibit the proliferation, migration and invasion of KB cells, and can also achieve apoptosis by inhibiting the activation of PI3K/Akt pathway. Therefore, TQ is an effective candidate for the treatment of OSCC.

Background: We aimed to investigate the effect of PI3K/Akt signaling pathway on PRAS40Thr246 phosphorylation in gastric cancer cells. Methods: The study was conducted from April 2017 to January 2018 in Zhongshan Hospital, Xiamen University, Xiamen, China. Gastric cancer cells were divided into three groups: gastric cancer cell group, LY294002 group and MK-2206 group. Specific tests were conducted accordingly. Results: Inhibition of PI3K/Akt signaling pathway activation and PRAS40Thr246 phosphorylation could inhibit proliferation and invasion and promote apoptosis of gastric cancer cells, and PRAS40Thr246 phosphorylation could activate PI3K/Akt signaling pathway. Conclusion: The levels of PI3K/Akt signaling pathway related proteins and p-PRAS40Thr246 were significantly increased in gastric cancer cells. p-PRAS40-Thr246 was able to reflect the activation of the PI3K/Akt signaling pathway, reflecting the sensitivity of the PI3K/AKT signaling pathway to inhibitors.

Glaucoma represents a major cause of blindness, generally associated with elevated intraocular pressure (EIOP). The aim of the present study was to investigate whether microRNA-149 (miR-149) affects retinal ganglion cells (RGCs) and the underlying mechanism based on a mouse model of chronic glaucoma with EIOP. The successfully modeled mice were administered with mimics or inhibitors of miR-149. Next, the number of RGCs, ultrastructural changes of RGCs, and purity of RGCs in the retinal tissues were detected. Moreover, the RGCs were collected and subsequently treated with 60 mmHg pressure and transfected with a series of plasmids aiding in the regulation of the expression of miR-149 and betacellulin (BTC). The levels of miR-149, BTC, phosphatidylinositol 3-kinase (PI3K), and Akt were subsequently determined. Finally, RGC viability and apoptosis were detected accordingly. Dual luciferase reporter gene assay provided validation, highlighting BTC was indeed a target gene of miR-149. The downregulation of miR-149 is accompanied by an increased number of RGCs and decreased ultrastructural RGC alterations. Additionally, downregulated miR-149 was noted to increase the levels of BTC, PI3K, and Akt in both the retinal tissues and RGCs, whereas the silencing of miR-149 was observed to promote the viability of RGC and inhibit RGC apoptosis. Taken together, the results of the current study provided validation suggesting that the downregulation of miR-149 confers protection to RGCs by means of activating the PI3K/Akt signaling pathway via upregulation of BTC in mice with glaucoma. Evidence presented indicated the promise of miR-149 inhibition as a potential therapeutic strategy for glaucoma treatment.

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Hepatocellular carcinoma (HCC) has become the third leading cause of death from cancer worldwide, and PI3K/AKT signaling pathway acts as the most common oncogenic pathway in HCC. But few studies have reported the prognostic value of PI3K/AKT associated genes (PAGs) and their association with immune infiltration. Hence, we downloaded the mRNA sequencing data and clinical information from The Cancer Genome Atlas (TCGA), GSE14520 dataset of Gene Expression Omnibus (GEO) and International Cancer Genome Consortium (ICGC) database. The 105 PAGs gene sets were from the Gene set enrichment analysis (GSEA) website. The biological processes of differently expressed PAGs were explored by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Then 4 prognostic PAGs (SFN, PRKAA2, PITX2 and CDK1) were identified through univariate and multivariate analyses. A prognostic signature was built base on the four PAGs. Afterward, the high-risk group had shorter survival time in Kaplan-Meier (KM) curves. Receiver operating characteristic (ROC) curves showed a better prognostic value of risk score (ROC=0.736) compared with other clinicopathological characteristics (AUC ≤ 0.511). The consistent results were obtained in the testing groups involving the GSE14520 dataset and ICGC database. The nomogram predicted the 1-year, 3-year, and 5-year overall survival rates in HCC. The correlation among risk score and immune infiltration of monocytes and M0 macrophages were determined, the expression levels of immune checkpoints (PDCD1, CTLA4, TIM3 and TIGIT) were also related to risk score in HCC. The study provided novel insight into the new targets for immunotherapy in HCC.

Renal cell carcinoma (RCC) is one of the most prevalent cancers. Long noncoding RNAs (LncRNAs) have been indicated as a mediator acted in tumorigenesis of RCC. However, the mechanism of LINC00460 on RCC is yet to be investigated. This study aimed to investigate the potential function of LINC00460 and underlying mechanism of RCC. We detected LINC00460 expression in RCC tissues and the prognosis in RCC patients using Gene Expression Profiling Interactive Analysis (GEPIA) website and The Cancer Genome Atlas (TCGA) database. LINC00460 level in normal renal cell line and RCC cell lines were detected by quantitative real-time polymerase chain reaction (qRT-PCR). We study the effects of LINC00460 on proliferation, migration, invasion, apoptosis in RCC cells lines using a series of in vivo and in vitro experiments. RNA sequencing (RNA-seq) analysis for the whole transcriptome was applied to searching potential LINC00460 related signal pathway in RCC. We identified the significant up-regulated expression level of LINC00460 in RCC tissues and cell lines. Elevated LINC00460 was correlated with shorter survival of RCC patients. Overexpression of LINC00460 promoted cell viability, proliferation, invasion and migration, while down-regulation of LINC00460 exerted inhibitory effect on these activities. We crucially identified that LNC00460 promotes development of RCC by influencing the PI3K/AKT pathway. Knockdown of LNC00460 decreased the phosphorylation of AKT and mTOR. The key finding of our study provided a new evidence suggesting that LINC00460 functions as an oncogene in RCC pathogenesis by mediating the PI3K/AKT pathway, which may provide a new target for the treatment of RCC.

Background: Killer cell immunoglobulin-like receptor 2DL4 (KIR2DL4) is a transmembrane glycoprotein that is expressed by natural killer (NK) cells and certain subsets of T cells. It has been reported to serve an important role in the immune response. However, its expression profiles and function in solid tumor progression remain poorly defined.Methods: In the present study, using bioinformatics analysis, immunohistochemistry, immunoblotting, MTT assay, soft agar colony formation assay and a renal cell carcinoma (RCC) cell xenograft model in nude mice, we examined whether KIR2DL4 is expressed by RCC and its possible roles in RCC progression. Results: We confirmed that KIR2DL4 is overexpressed by RCC cells. MTT and soft agar cloning assays showed that KIR2DL4 knockdown delayed cell proliferation in RCC cell lines, Caki-1 and 769-P, in vitro. By contrast, KIR2DL4 overexpression promoted Caki-1 cell proliferation both in vitro and in vivo, which was observed in a BALB/c-nu/nu xenograft mouse model. Moreover, RNA sequencing data demonstrated that the differentially expressed genes between vector controlled and KIR2DL4-overexpressed Caki-1 cells were highly associated with cancer development, of which those related to the phosphatidylinositol-3-kinase (PI3K)/ protein kinase B (AKT) signaling pathway were particularly enriched. Immunoblotting data showed that the level of AKT phosphorylation was higher in KIR2DL4-overexpressing Caki-1 cells compared with that in the parallel-controlled cells. Conclusions: Our results indicate that KIR2DL4 is also expressed by RCC cells, which promotes RCC progression through the PI3K/AKT signaling pathway.

Background Esophageal cancer remains a dominating cause of cancer-associated death and has shown a sharp increase of more than 6-fold increase rates all over the world. It is divided into two main pathological types: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). Most of the patients are diagnosed at the advanced stage with distant metastasis and poor prognosis. Thus, it is urgent for us to identify new more effective drugs to improve the prognosis of patients with esophageal cancer. Elaiophylin is a novel autophagy inhibitor at the late stage, which is a C2 symmetric macrolides separated from Streptomyces Niger. It was proved to have anti-tumor ability of various of cancers, yet little is known whether it could inhibit the progression of esophageal squamous cancer. Methods The protein expression level was detected by western blot assay. The viability of ESCC cells was detected by CCK8 assay. The proliferation rate was measured by ki67 immunofluorescent staining. The migratory ability of ESCC cells was identified by transwell assay. The apoptosis rate of ESCC cells was explored by Annexin V/PI analysis. And the genes might be regulated by Elaiophylin was analyzed by RNA-seq assay and verified by RT-qPCR assay. Results In the study, we firstly found that the Elaiophylin was a novel autophagy inhibitor in ESCC cells which is consistent with the previous studies. In addition, Elaiophylin can inhibit the proliferation and migration of ESCCs, and accelerate its apoptosis. And the activity of PI3K/AKT pathway was inhibited by Elaiophylin, the mRNA and protein expression level of EIF4B was both downregulated by Elaiophylin. Mechanistically, silencing of EIF4B could inhibit the proliferation and migration of ESCC cells and promoted apoptosis, which could be rescued by EIF4B-OE. Moreover, EIF4B overexpression suppressed the apoptosis, while increased the proliferation and migration of ESCC cells which resulted from Elaiophylin. Conclusions Together, our results indicated that Elaiophylin is a novel autophagy inhibitor governing growth and migratory ability of ESCC cells through PI3K/AKT signaling pathway. simultaneously, and identified EIF4B as a target of Elaiophylin, which might provide a new therapeutic strategy for esophageal squamous carcinoma.

IntroductionDysregulated angiogenesis is a critical characteristic for endothelial dysfunction disorders. This study aimed to determine functions of microRNA-126 in formation of tube and investigated the potential mechanisms.Material and methodsThe synthesized microRNA-126 control and microRNA-126 inhibitor plasmids were transfected into human umbilical-vein endothelial cells (HUVECs) using lipofectamine 2000 reagent. Cell counting kit-8 (CCK-8) was employed to measure proliferative capability of HUVECs. Transwell analysis was used to evaluate HUVECs invasive ability. Real time PCR (RT-PCR) was utilized to access epidermal growth factor-like domain 7 (EGFL7) and microRNA-126 mRNA transcription. Tube-forming capability in HUVECs was determined. Dual-luciferase assay and linear-regression analysis were conducted to measure interaction between EGFL7 and microRNA-126 molecule. Phosphoinositide-3-kinase/protein kinase-B (PI3K/AKT) signaling pathway associated molecules were evaluated using western blot assay.ResultsSilencing of microRNA-126 significantly enhanced proliferative capability and invasive ability of HUVECs compared to those of microRNA-126 control group (pConclusionsmicroRNA-126 modulated angiogenesis and tube formation through increasing EGFL7 expression and phosphorylating PI3K/AKT signaling pathway.