miR-182靶向NUMB调控卵巢癌SKOV3细胞增殖和侵袭的机制研究

Effects of miR-182 targeting NUMB on proliferation and invasion of ovarian cancer SKOV3 cells

目的研究miR-182在卵巢癌SKOV3细胞中的表达以及对卵巢癌细胞增殖和侵袭的影响。方法运用qRT-PCR检测miR-182和NUMB在人卵巢癌上皮细胞系SKOV3和人卵巢正常上皮细胞系HOSE中的表达,将SKOV3细胞分为Control组(未转染)、anti-NC组(转染无意义序列)以及anti-miR-182组(转染anti-miR-182)。MTT法检测各组SKOV3细胞的增殖;Transwell实验检测各组SKOV3细胞的侵袭能力;Western blot检测抑制miR-182表达对NUMB表达的影响;生物学网站分析及双荧光素酶报告基因实验研究miR-182和NUMB的靶向关系。结果和人卵巢正常上皮细胞系HOSE相比,卵巢癌细胞SKOV3中的miR-182表达升高,NUMB的表达降低(P<0.05)。MTT和Transwell实验结果显示,抑制miR-182的表达可以抑制卵巢癌细胞的增殖和侵袭(P<0.05)。抑制卵巢癌细胞SKOV3中miR-182的表达,可以促进NUMB的表达,且在NUMB基因3′UTR区存在miR-182的结合位点,双荧光素酶报告基因实验结果提示NUMB是miR-182的靶基因。结论miR-182在卵巢癌细胞SKOV3中的表达升高,抑制miR-182表达可以抑制卵巢癌细胞SKOV3的增殖及侵袭能力,机制可能靶向负调控NUMB的表达有关。

Objective To study the expression of miR-182 in ovarian cancer cells and its effect on the proliferation and invasion of ovarian cancer cell SKOV3.Methods The expressions of miR-182 and NUMB in human ovarian cancer SKOV3 cell and normal human ovarian epithelial HOSE cell were detected by qRT-PCR.The anti-miR-182 was transfected into SKOV3 cells to inhibite expression of miR-182.MTT method was used to detect proliferation of cells.Transwell assay was used to detect invasion of cells.Western blot was used to detect the expression of NUMB in each group.Biological website analysis and double luciferase reporter gene experiment were used to study the targeting relationship between miR-182 and NUMB.Results Compared with human normal ovarian epithelial cell HOSE,the expression of miR-182 was increased in ovarian cancer cells SKOV3,and the expression of NUMB was decreased.Compared with the control group,the proliferation and invasion ability of ovarian cancer cells in anti-miR-182 group was inhibited(P<0.05).Transfection of anti-miR-182 can increase the expression of NUMB in ovarian cancer cells.There is a binding site of miR-182 in the 3′UTR region of NUMB gene.Luciferase analysis showed that NUMB was a target gene of miR-182.Conclusion The expression of miR-182 was increased in ovarian cancer cell SKOV3.Inhibiting the expression of miR-182 can inhibit the proliferation and invasion of ovarian cancer cell SKOV3,which may be related to the negative regulation of NUMB expression.