长链非编码RNA MEG3靶向Rac1抑制宫颈癌细胞增殖、迁移的实验研究

Long chain non-coding RNA MEG3 targeting Rac1 inhibits proliferation and migration of cervical cancer cells

目的检测长链非编码RNA MEG3在宫颈癌细胞中的表达,检测MEG3表达对He La细胞增殖、迁移的影响及可能分子机制。方法采用荧光定量PCR(QPCR)检测正常宫颈上皮细胞Hcer Epic和宫颈癌细胞系C-33A、C4-1、Caski、Si Ha和He La中MEG3的表达; MEG3过表达质粒(MEG3)、阴性对照质粒(NC组)、MEG3+Rac1过表达质粒(MEG3+Rac1组)分别转染He La细胞,MTT法检测细胞增殖情况,Transwell实验检测细胞迁移能力,Western blotting检测PCNA、E-cadenin、N-cadenin、Vimentin、Rac1蛋白表达。结果 QPCR结果显示,C-33A、C4-1、Caski、Si Ha和He La细胞中MEG3表达水平分别为0. 37±0. 044、0. 65±0. 075、0. 41±0. 071、0. 71±0. 053和0. 42±0. 081,均低于Hcer Epic细胞的1. 02±0. 064,差异具有统计学意义(P<0. 05)。MTT法结果显示MEG3组He La细胞增殖活力显著低于NC组; MEG3组的迁移细胞数为385±14,显著低于NC组的594±16(P<0. 05);与NC组比较,MEG3组PCNA、N-cadenin和Vimentin表达下调,E-cadenin表达上调。与MEG3组比较,MEG3+Rac1组显著上调Rac1蛋白表达,上调He La细胞增殖活力,增加迁移细胞数。结论 LncRNA MEG3可能通过靶向Rac1信号通路抑制宫颈癌细胞增殖、迁移。

Objective To detect the expression of long chain non-coding RNA MEG3 in cervical cancer cells,and to detect the effect of MEG3 on the proliferation and migration of HeLa cells and its possible molecular mechanism.Methods The expression of MEG3 in normal cervical epithelial cells HcerEpic and cervical cancer cell lines C-33A,C4-1,Caski,SiHa and HeLa was detected by fluorescence quantitative PCR(QPCR).MEG3 overexpression plasmid(MEG3 group),negative control plasmid(NC group)and MEG3+Rac1 overexpression plasmid(MEG3+Rac1 group)were transfected into HeLa cells respectively.MTT assay was used to detect cell proliferation and Transwell assay was used to detect cell migration.The expression of PCNA,E-cadenin,N-cadenin,Vimentin and Rac1 was detected by Western blotting assay.Results The expression levels of MEG3 in C-33A,C4-1,Caski,SiHa and HeLa cells were 0.37±0.044,0.65±0.075,0.41±0.071,0.71±0.053 and 0.42±0.081,which were lower than those in HcerEpic cells(P<0.05).MTT assay showed that the proliferation activity of HeLa cells in MEG3 group was significantly lower than that in NC group.The number of migrating cells in MEG3 group was 385±14,which was significantly lower than that in NC group(594±16),and the difference was statistically significant(P<0.05).Compared with NC group,the expressions of PCNA,N-cadenin and Vimentin were down-regulated and E-cadenin was up-regulated in MEG3 group.Compared with MEG3 group,MEG3+Rac1 group significantly increased the expression of Rac1 protein,the proliferation activity and the number of migrating cells.Conclusion LncRNA MEG3 inhibits the proliferation and migration of cervical cancer cells by targeting Rac1 signaling pathway.