miR-539通过靶向EMT调节宫颈癌细胞增殖、转移

miR-539 regulates the proliferation and metastasis of cervical cancer cells by targeting EMT

目的 探讨微小RNA-539(miR-539)对人宫颈癌细胞C33a增殖、转移的影响及转移机制。方法 收集宫颈癌组织及癌旁正常组织并检测组织中miR-539 mRNA含量;以人正常宫颈上皮细胞系HUCECs作为对照,筛选宫颈癌细胞中miR-539 mRNA相对表达量最低的细胞株;甲基噻唑基四唑(MTT)法检测宫颈癌细胞的增殖;Transwell小室法检测miR-539对细胞侵袭、迁移能力的影响;实时定量聚合酶链反应(RT-qPCR)检法、蛋白免疫印迹(Western blot)法检测波形蛋白(vimentin)相对表达量。结果 RT-qPCR结果显示,宫颈癌癌组织中miR-539含量较癌旁正常组织显著减少(P<0.05);与人正常宫颈上皮细胞系HUCECs相比,宫颈癌细胞C33a的miR-539明显降低(P<0.05);当成功过表达C33a细胞中miR-539后,与空白对照组相比,mimics-miR-539组增殖能力减弱(P<0.05);与空白对照组相比,过表达miR-539组细胞侵袭和迁移能下降(P<0.05);mimics-miR-539组vimentin mRNA及蛋白相对表达量较对照组显著降低(P<0.05)。结论 miR-539在人宫颈癌表达降低,在人宫颈癌细胞C33a中,过表达miR-539能够抑制其增殖、迁移,迁移机制可能与下调vimentin表达有关。

Objective To investigate the effect of microRNA-539(miR-539) on the proliferation and metastasis of human cervical cancer cell C33a and its mechanism of metastasis. Methods Collect cervical cancer tissues and normal tissues adjacent to the cancer and detect the level of miR-539 mRNA in the tissues. Human normal cervical epithelial cell line HUCECs were used as controls to screen the relative expression of miR-539 mRNA in cervical cancer cells the lowest cell line.Methyl thiazolyl tetrazolium method was used to detect the proliferation of cervical cancer cells. Transwell chamber method was used to detect the effect of miR-539 on cell invasion and migration. Real-time quantitative polymerase chain reaction(RT-qPCR) method and Western blot method were used to detect the relative expression of vimentin. Results The content of miR-539 in cervical cancer tissues was significantly lower than that in normal tissues adjacent to the cancer(P<0.05). After successfully overexpressing miR-539 in C33a cells, compared with human normal cervical epithelial cell line HUCECs, the miR-539 of cervical cancer cell C33a was significantly reduced(P<0.05). Compared with the blank control group, the proliferation ability of the mimics-miR-539 group was weakened(P<0.05). Compared with the blank control group, the cell invasion and migration energy of the up-regulated miR-539 group decreased(P<0.05). The relative expression of vimentin mRNA and protein in the mimics-miR-539 group was significantly lower than that in the control group(P<0.05). Conclusion The expression of miR-539 is decreased in human cervical cancer. In human cervical cancer cell C33a, overexpression of miR-539 can inhibit its proliferation and migration and the migration mechanism may be related to the down-regulation of vimentin expression.