摘要
目的 探讨lncRNA母系表达基因3(MEG3)经miR-421调节E-上皮钙黏附素(E-cadherin)抑制乳腺癌细胞上皮间充质转化(EMT)的机制。方法 qRT-PCR法检测45例乳腺癌及癌旁组织lncRNA MEG3 mRNA表达。通过生物信息学网站筛选靶向lncRNA MEG3的miRNA。乳腺癌MCF-7细胞分别转染miR-NC、lncRNA MEG3 RNAi和pCDNA3.0-HA-lncRNA MEG3质粒,qRT-PCR检测各组细胞lncRNA MEG3、miR-421和E-cadherin mRNA表达水平;MTT法、Transwell法检测细胞增殖和迁移情况。结果 乳腺癌组织lncRNA MEG3表达低于癌旁组织(P<0.05)。lncRNA MEG3与miR-421存在靶向调节作用。与阴性对照组比较,细胞lncRNA MEG3和E-cadherin mRNA表达在MEG3 RNAi组降低,MEG3质粒组增加;miR-421表达、细胞增殖率和迁移数在MEG3 RNAi组增加,MEG3质粒组降低(P<0.05)。结论 lncRNA MEG3通过miR-421靶向调控E-cadherin,抑制乳腺癌细胞EMT。
Aim To explore the mechanism of lncRNA maternal expressed gene3(MEG3) regulating E-cadherin by microRNA-421(miR-421) to inhibit epithelial-mesenchymal transition(EMT) of breast cancer cells. Methods qRT-PCR method was used to detect the expression level of lncRNA MEG3 mRNA in 45 cases of breast cancer and adjacent tissues. The miRNAs targeting lncRNA MEG3 were predicted and screened through the bioinformatics website. The miR-NC, lncRNA MEG3 RNAi and pCDNA3.0-HA-lncRNA MEG3 plasmid was transfected into MCF-7 cells by Lipofectamine method, and after 48 hours of continuous culture, the expression levels of lncRNA MEG3, miR-421 and E-cadherin mRNA in the cells were detected by qRT-PCR method, and the proliferation and migration of cells were detected by Methyl tihiazolyl tetrazolium(MTT) method and Transwell method respectively. Results The expression level of lncRNA MEG3 in breast cancer tissues was significantly lower than paired adjacent tissues(P<0.05). The lncRNA MEG3 had a potential binding site with miR-421 and had a targeted regulatory effect. Compared with the negative control group, the expression of lncRNA MEG3 and E-cadherin mRNA in MCF-7 cells decreased in the MEG3 RNA group and increased in the MEG3 plasmid group, and the expression of miR-421, cell proliferation rate and migration number increased in the MEG3 RNAi group and decreased in the MEG3 plasmid group(P<0.05). Conclusion LncRNA MEG3 regulates E-cadherin by targeting miR-421 and inhibits EMT in breast cancer cells.
作者
王清云
王秀艳
尹婷婷
胡爽
WANG Qingyun;WANG Xiuyan;YIN Tingting;HU Shuang(Clinical Laboratory,the First Affiliated Hospital of Xingtai Medical College,Xingtai First Hospital,Xingtai 054001,Hebei,China)
出处
《中南医学科学杂志》
CAS
2023年第1期41-44,共4页
Medical Science Journal of Central South China
基金
邢台市重点研发计划项目(2020ZC198)。
