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【Objective】 To investigate the role of microRNA-218 (miR-218) in regulating prostate cancer (PCa) cell stemness and epithelial-mesenchymal transition (EMT). 【Methods】 PCa cell line stably overexpressing miR-218 was constructed with lentivirus transfection. The expression of miR-218 was detected with real-time fluorescence quantitative polymerase chain reaction (q-PCR). The migration ability was detected with Transwell assay. The expression of EMT related proteins were detected with Western blot. The properties of cells were determined with colony formation and tumor sphere formation assays. 【Results】 The results of q-PCR showed that the mRNA level of miR-218 was significantly lower in PCa cell lines LNCaP and C4-2 than in BPH-1. Transwell assay showed that miR-218 inhibited the migration of PCa cells. Western blot showed that the expression of EMT related proteins were inhibited by miR-218. Colony formation and tumor sphere formation assays showed that overexpression of miR-218 significantly inhibited the properties of cells. 【Conclusion】 The expression of miR-218 is downregulated in PCa cell lines. miR-28 can inhibit cell migration, EMT and cancer stem cell properties.
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AIM: To investigate the protective effect of microRNA-218 (miR-218) silencing on kidney tissue of streptozotocin (STZ)-induced diabetic nephropathy rats and the potential mechanism.METHODS: The diabetic rat model was established by a single intraperitoneal injection of STZ (50 mg/kg).Meanwhile, the miR-218 short hairpin RNA (shRNA) lentiviral vector was constructed.The Sprague-Dawley rats were randomly divided into 4 groups: healthy control group, diabetes group, empty vector group and miR-218-shRNA group.The blood glucose, 24 h urinary protein, serum creatinine (SCr) and blood urea nitrogen (BUN) in the rats at different time points (4, 8 and 12 weeks) were measured by an automated analyzer.The expression of miR-218 was detected by RT-qPCR, while the expression of heme oxygenase-1 (HO-1), nephrin and p38 mitogen-activated protein kinase (p38 MAPK) at mRNA and protein levels in the kidney tissues was determined by RT-qPCR and Western blot.The caspase-3 activity was detected by caspase-3 activity assay kit, and the cell apoptosis of the kidney tissues was analyzed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).RESULTS: Compared with healthy control group, the expression of miR-218 was significantly increased in STZ-treated rats.Meanwhile, the concentrations of blood glucose, 24 h urinary protein, SCr and BUN were significantly increased in STZ-treated rats (P<0.05).The mRNA and protein expression of HO-1 and nephrin was significantly decreased, while the level of phosphorylated p38 MAPK was significantly increased in STZ-treated rats.In addition, the activity of caspase-3 was also significantly increased in STZ-treated rats.When the model rats were infected with miR-218-shRNA, the expression of miR-218 was significantly decreased and the above effects were markedly reversed.Furthermore, TUNEL results showed that compared with diabetic group and empty vector group, miR-218 silencing significantly attenuated the cell apoptosis in the kidney tissues in miR-218-shRNA group.CONCLUSION: miR-218 is involved in the kidney injury in diabetic rats, and silencing of miR-218 by lentiviral vector-mediated miR-218-shRNA transfection effectively inhibits kidney cell apoptosis, suggesting that miR-218 is a potential target for the treatment of diabetic nephropathy.
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Objective To investigate the expression of miR-218 (microRNA-218) in breast cancer tissues and its clinical significance. Methods Totally 45 tissue biopsies gained from breast cancer patients and the adjacent normal tissue were collected. Real-time qPCR was used to detect the miR-218 expressions. The correlations of miR-218 expression with clinicopathological characteristics and prognosis of breast cancer patients were analyzed. Results The average expression level of miR-218 in breast cancer tissues was significantly lower than that in control tissues (P = 0.001). The average expression level of miR-218 in PR negative breast cancer tissues was significantly lower than that of PR positive breast cancer tissue (P = 0.037). The average expression level of miR-218 in Ki-67 positive breast cancer tissues was significantly lower than that in Ki-67 negative breast cancer tissue (P=0.018). Conclusion The expression of miR-218 is closely related to carcinogenesis ,progres-sion and prognosis of breast cancer ,so it may be served as a diagnostic biomarker and a prognostic predictor in breast cancer patients,which provides a new way for the treatment of breast cancer.
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Objective To study the expression level and function of micro RNA (microRNA)-218 in hepatocellular carcinoma (HCC) .Methods 46 cases of HCC surgery in the hepatobiliary surgery department of this hospital were selected and divided into the transfection group and nontransfection group .The expression ,proliferation and apoptosis of microRNA-218 and the expression level of B cell specific Maloney leukemia virus insertion site 1(Bmi-1) and cycling-dependented kinase 6(CDK6) in HepG2 cells were compared between the two groups .Results The expression level of microRNA-218 in HCC tissue was significantly lower than that in paracancerous tissues (P＜0 .05);the microRNA218 expression level was closely correlated with the clinicopathological characteristics such as tumor size and TNM stage(P＜0 .05);the HepG2 cell proliferation rates at 24 ,48 ,72 h after transfection in the transfection group were significantly lower than those in the nontransfection group(P＜0 .05);the HepG2 cell apoptosis rate in the transfection group was significantly higher than that in the nontransfection group(P＜0 .05);the Bim-1 and CDK6 expression levels after HepG2 cell transfection in the transfection group were significantly lower than those in nontransfection group(P＜0 .05) . Conclusion microRNA-218 can suppress the proliferation of HCC cells and promotes HCC cells apoptosis by down-regulating the Bim-1 and CDK6 expression level in potential targets .