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[摘要]
目的:探讨lncRNA SBF2-AS1 通过调控miR-140-5p/血管内皮生长因子A(VEGFA)分子轴对宫颈癌HeLa 细胞上皮间质转化(EMT)的影响。方法:细胞培养和转染后分为NC、miR-140-5p mimic、miR-140-5p mimic+pcDNA-VEGFA、si-lncRNASBF2-AS1+pcDNA-VEGFA及si-lncRNA SBF2-AS1+miR-140-5p mimic组5 组。采用qPCR检测lncRNA SBF2-AS1 在宫颈癌组织及细胞系中的表达水平,双荧光素酶报告基因验让lncRNA SBF2-AS1、miR-140-5p 与VEGFA的靶向关系,WB检测HeLa细胞中VEGFA及EMT标志物N-cadherin、Vimentin 和E-cadherin 的表达水平,Transwell 实验检测HeLa细胞侵袭和迁移能力。结果:lncRNASBF2-AS1 在宫颈癌组织及细胞系中高表达(P<0.05 或P<0.01),lncRNA SBF2-AS1 靶向结合miR-140-5p,且VEGFA 是miR-140-5p 的靶基因(P<0.05)。敲降lncRNA SBF2-AS1 抑制HeLa细胞侵袭、迁移及EMT。进一步实验证实,lncRNA SBF2-AS1通过miR-140-5p 上调VEGFA的表达水平,从而促进HeLa 细胞侵袭、迁移及EMT(P<0.05 或P<0.01)。结论:lncRNA SBF2-AS1通过miR-140-5p/VEGFA分子轴促进HeLa细胞EMT。
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[Abstract]
Objective: To investigate the effect of lncRNA SBF2-AS1 on epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cell via regulating miR-140-5p/VEGFA (vascular endothelial growth factor A) axis. Methods: After cell culture and transfection,the cells were divided into 5 groups: NC group, miR-140-5p mimic group, miR-140-5p mimic+pcDNA-VEGFA group, si-lncRNA SBF2-AS1+pcDNA-VEGFA group and si-lncRNA SBF2-AS1+miR-140-5p mimic group. The expression level of lncRNA SBF2-AS1 in cervical cancer tissues and cell lines was detected by qPCR. The targeted relationship between lncRNA SBF2-AS1, miR-140-5p and VEGFA was confirmed by Dual luciferase reporter gene assay. The expression levels of VEGFA and EMT-related proteins N-cadherin,Vimentin and E-cadherin in HeLa cells were detected by WB. The invasion and migration of HeLa cells were detected by Transwell.Results: lncRNA SBF2-AS1 was highly expressed in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Dual luciferase reporter gene assay confirmed that lncRNA SBF2-AS1 targetedly combined with miR-140-5p and VEGFA was a target gene of miR-140-5p (P<0.05). Knockdown of lncRNA SBF2-AS1 inhibited invasion and migration as well as EMT of HeLa cells. Further experiment confirmed that lncRNA SBF2-AS1 up-regulated the expression level of VEGFA via miR-140-5p, thereby promoting invasion, migration and EMT of HeLa cells. Conclusion: lncRNA SBF2-AS1 promotes EMT of HeLa cells via miR-140-5p/VEGFA axis.
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