| MiR-107(microRNA-107)is located on human chromosome 10q23.31 and is a region where the chromatin frequently changes.CHUNG et al found that miR-107 in endometrial adenocarcinoma of the endometrium,pancreatic cancer,acute promyelocytic leukemia in much higher than normal tissue.Lee et al.Found that miR-107 functions as a tumor suppressor gene in pancreatic cancer cells through methylation of miR-107 promoter and inhibition of cyclin-dependent kinase 6(CDK6)levels.However,there is no further study on miR-107 in endometrial carcinoma.Therefore,we want to study the role and possible mechanism of miR-107 in human endometrial adenocarcinoma.In this study,Ishikawa cell transfected with miR-107 inhibitor reagent in human endometrial adenocarcinoma in vitro was found to significantly downregulate the expression of miR-107 in Ishikawa cells of human endometrial adenocarcinoma,meanwhile,the proliferation,migration and invasion of cells was also decreased.In conclusion,the transfection of miR-107 inhibitor significantly downregulated the expression of miR-107 in Ishikawa cells of human endometrial adenocarcinoma.Down-regulation of miR-107 may be related to upregulation of ER-α expression to inhibit Ishikawa cell proliferation and migration and invasion.The First StepTransfection of miR-107 inhibitor significantly downregulated the expression of miR-107 in Ishikawa cells of human endometrial carcinoma.Objective:To research the effect of miR-107 inhibitor transfection in vitro on the expression of miR-107 in Ishikawa cells of human endometrial carcinomaMethods:The human endometrioid adenocarcinoma Ishikawa cell line purchased from Shanghai Cell Bank of Chinese Academy of Sciences was routinely cultured in vitro and the cells in logarithmic growth phase were tested.The experiment was divided into three groups:(A)untreated:(B)negative control group(miR-107i NC): transfected with miR-107 inhibitor negative control;(C)downregulation group(miR-107i): cells were transfected with miR-107 inhibitor.According to Lipofectamine (?) LTX instruction,miR-107 inhibitor and miR-107 inhibitor negative control were transfected into Ishikawa cells by Lipofectamin TM 3000 reagent at 37 ° C in 5% CO2 incubator for 48 h and detected by qRT-PCR The expression level of miR-107 in each group of Ishikawa cells.Results:The results of qRT-PCR showed that there was no difference in miR-107 expression between untreated group and Ishikawa cells in miR-107i NC group(P=0.790,P>0.05).Compared with miR-107i NC group,the expression of miR-107 in Ishikawa cells of miR-107i group was significantly decreased(P=0.0049,P<0.01).Compared with untreated group,the expression of miR-107 in Ishikawa cells of miR-107i group was also decreased(P=0.011,P<0.05).Conclusion : Transfection of miR-107 inhibitor in vitro could significantly down-regulate the expression of miR-107 in Ishikawa cells of human endometrial carcinoma.The Second Step Down-regulation of miR-107 expression can inhibit the proliferation,migration and invasion ability of human Ishikawa cells.Objective:To research the effect of down-regulation of miR-107 expression on proliferation,migration and invasion of Ishikawa cells of human endometrial carcinomaMethods:The logarithmic growth phase of Ishikawa cells that had a significantly lower expression of miR-107 after transfection with miR-107 inhibitor was routinely cultured.The cell absorbance at 490 nm in each group at 24 h,48h and 72 h was detected by CCK8 assay.The migration ability of cells in each group at 24 h,48h and 72 h was detected by wound healing assay.The invasion ability of cells of each group at 48 h was detected by Transwell migration assay.Results:(1)CCK8 assay results showed that there was no difference in OD value between untreated group and miR-107i NC group at 24 h,48h and 72 h.(24h:P=0.967,P>0.05;48h: P=0.219,P>0.05;72h:P=0.069,P>0.05).Compared with miR-107i NC group,Ishikawa cells in miR-107i group at 24 h,48h and 72 h OD values were significantly lower(24h,48h和72h均P<0.0001).Compared with untreated group,Ishikawa cells in miR-107i group at 24 h,48h and 72 h OD values were also significantly lower(24h,48h和72h均P<0.0001).(2)Wound healing assay showed that there was no difference in Ishikawa cells between untreated group and miR-107i NC group at 24 h,48h and 72h(P=0.641,P>0.05;P=0.107,P>0.05;P=0.080,P>0.05).Compared with miR-107i NC group, there was no difference in Ishikawa cell migration in miR-107i group at 24h(P=0.085,P>0.05)Ishikawa cells but decreased at 48 h and 72h(48h:P=0.010,P<0.05;72h:P=0.013,P<0.05).Compared with untreated group,there was no difference in Ishikawa cell migration in miR-107i group at 24h(P=0.062,P>0.05)Ishikawa cells but decreased at 48 h and 72h(48h:P=0.013,P<0.05;72h:P=0.028,P<0.05).(3)Transwell migration assay results showed that there was no difference in the number of cells crossing the matrigel between untreated group and miR-107i NC group(P=0.950,P>0.05).The number of cells passing through the matrigel in the miR-107i group was significantly reduced compared with the miR-107i NC group(P=0.005,P<0.01).The number of cells passing through the matrigel in the miR-107i group was also reduced compared with untreated group(P=0.005,P<0.01)Conclusion:Down-regulation of miR-107 expression can inhibit the proliferation,migration and invasion ability in Ishikawa cells of human endometrial carcinoma.The Third Step Down-regulation of miR-107 expression can promote the expression of ER-α in Ishikawa cells of human endometrial carcinoma.Objective:To research the effect of down-regulation of miR-107 expression in vitro on the expression of ER-α in Ishikawa cells of human endometrial carcinoma.Methods:The logarithmic growth phase of Ishikawa cells that had a significantly lower expression of miR-107 after transfection with miR-107 inhibitor was routinely cultured.Western blotting was used to detect the expression of ER-α in Ishikawa cells of endometrial carcinoma.Results:The results of western blotting analysis showed that there was no difference in the expression of ER-α between untreated group and miR-107i NC group(P=0.989,P>0.05).Compared with miR-107i NC group,the expression of ER-α in Ishikawa cells of miR-107i group increased(P=0.016,P<0.05).Compared with untreated group,the expression of ER-α in Ishikawa cells of miR-107i group also increased(P=0.022,P<0.05).Conclusion:.Down-regulation of miR-107 expression can promote the expression of ER-α in Ishikawa cells of human endometrial carcinoma. |
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