Effect of miR-599 expression changes on proliferation and invasion of ovarian cancer cells / 中国医师杂志

ABSTRACT

Objective:To investigate the expression of microRNA (miR)-599 in ovarian cancer and its effect on the proliferation and invasion of ovarian cancer cells.Methods:Quantitative real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression of miR-599 in 36 cases of ovarian carcinoma and paracancerous tissues, ovarian cancer cell lines and normal ovarian epithelial cells. The miR-599 inhibitor (experimental group) and the negative control miR-NC (control group) were transfected into the ovarian cancer cell line A2780, respectively. qPCR was used to detect the transfection efficiency. Bioinformatics technology was used to predict and screen candidate target genes of miR-599, and dual luciferase reporter gene analysis system was established to verify the direct targeting effect of miR-599 on candidate target genes. qPCR and Western blot were used to detect the mRNA and protein expression changes of target genes. Cell counting (CCK-8) and transwell invasion assays were used to detect the effect of miR-599 on proliferation and invasion of A2780 cells.Results:qPCR results showed that the expression of miR-599 in ovarian cancer tissues was higher than that in paracancerous tissues ( P<0.01). The expression level of miR-599 in ovarian cancer cells was higher than that in normal ovarian epithelial cells ( P<0.05). The opioid binding protein (OPCML) is a candidate target gene for miR-599. The dual luciferase reporter assay confirmed that miR-599 can directly target the 3′-untranslated region (3′-UTR) of the OPCML gene ( P<0.01). After down-regulation of miR-599 expression, the mRNA and protein expression levels of OPCML in A2780 cells decreased ( P<0.01); the expression of cell cycle regulatory proteins such as CDK4 and cyclin D2 was decreased; the expression of negative regulatory protein E-cadherin was increased, and the expression of positive regulatory protein N-cadherin was decreased; the proliferation and the invasion ability of A2780 cells decreased ( P<0.05). Conclusions:miR-599 is highly expressed in ovarian cancer tissues and cell lines. Down-regulation of miR-599 expression in ovarian cancer cells A2780 can inhibit the proliferation and invasion of ovarian cancer cells by increasing OPCML gene expression.