miR-145对宫颈癌细胞增殖、迁移和侵袭的影响

Effects of miR-145 on proliferation,migration and invasion of cervical cancer cells

目的研究miR-145对人宫颈癌细胞系SiHa细胞增殖、迁移和侵袭的影响,并探讨其机制。方法体外培养子宫颈癌细胞和正常宫颈上皮细胞,检测细胞中miR-145表达水平;将SiHa细胞分为miR-145过表达(mimics)组和阴性对照(NC)组,转染48 h后,CKK-8法检测细胞增殖情况,Transwell实验检测细胞迁移、侵袭能力;应用TargetScan数据库预测miR-145的靶基因并用荧光素酶报告基因实验验证,蛋白免疫印记(WB)检测miR-145过表达对肌球蛋白Ⅵ(MY06)蛋白表达的影响。结果 SiHa、Hela、MS751人子宫颈癌细胞中miR-145表达水平均显著低于正常子宫颈上皮细胞,差异有统计学意义(P<0.05)。转染miR-145 mimics后,SiHa细胞中miR-145表达水平显著上调(P<0.05);与NC组比较,mimics组SiHa细胞的增殖、迁移、侵袭能力均显著下降(P<0.05)。TargetScan数据库预测显示MY06是miR-145潜在靶基因,双荧光素酶报告基因实验证实二者存在靶向关系。WB结果显示miR-145过表达后,MY06蛋白表达水平显著下调。结论 miR-145能通过下调MY06表达抑制人宫颈癌细胞系SiHa细胞的增殖、迁移和侵袭。

Objective To study the effect of miR-145 on the proliferation,migration and invasion of human cervical cancer SiHa cells and explore its mechanism. Methods The human cervical cancer cells and normal cervical epithelial cells were cultured,and the expression levels of miR-145 in the cells were detected. SiHa cells were divided into miR-145 overexpression(mimics)group and negative control(NC)group. 48 h post transfection,CKK-8 assay was used to detect cell proliferation. Transwell assay was used to detect cell migration and invasion abilities;the TargetScan database was used to predict the target of miR-145 and verified by the luciferase reporter gene assay. Protein immunoblotting(WB)were used to detect the effect miR-145 overexpression on the expression of myosin VI(MY06). Results The expression of miR-145 in cervical cancer cells of SiHa,Hela and MS751 were significantly lower than that in normal cervical epithelial cells(P<0.05). After transfection of miR-145 mimics,the expression of miR-145 in SiHa cells was significantly up-regulated(P<0.05);compared with the NC group,the proliferation,migration and invasiveness of SiHa cells in mimics group were significantly decreased(P<0.05). The TargetScan database prediction showed that MY06 is the potential target gene of miR-145,and the double luciferase reporter gene experiment confirmed that there is a target relationship between them. WB assay showed that the expression of MY06 was decreased significantly after the overexpression of miR-145.Conclusion MiR-145 could inhibit the proliferation,migration and invasion of human cervical cancer cell line SiHa cells by down-regulated the expression of MY06.