LncRNA HOTAIR对宫颈癌Hela细胞增殖、凋亡和EMT的影响和作用机制

Effects of LncRNA HOTAIR on proliferation,apoptosis and EMT of cervical cancer Hela cells and its potential mechanism

目的探讨长链非编码RNA(LncRNA)HOTAIR对宫颈癌Hela细胞增殖、凋亡和上皮细胞-间充质转化(EMT)的影响及潜在的作用机制。方法收集培养的宫颈癌细胞(Hela细胞)、人永生化宫颈上皮细胞(H8细胞),同时收集宫颈癌组织、癌旁正常组织标本,利用实时荧光定量PCR检测LncRNA HOTAIR、miR-20a-5p和KIF26B的mRNA表达;分别下调LncRNA HOTAIR、miR-20a-5p和KIF26B的表达,MTT试验检测Hela细胞增殖能力,Western blot检测Hela细胞内凋亡相关蛋白Bax、Bcl-2和EMT相关蛋白E-cadherin、N-cadherin的表达,流式细胞仪检测Hela细胞凋亡情况。miRanda和双荧光素酶报告基因试验分析LncRNA HOTAIR和miR-20a-5p之间的作用靶点及相关性,TargetScan和双荧光素酶报告基因试验分析miR-20a-5p与KIF26B之间的作用靶点及相关性;检测LncRNA HOTAIR通过miR-20a-5p对Hela细胞增殖、凋亡和EMT的影响。结果Hela细胞内LncRNA HOTAIR和KIF26B表达明显高于H8细胞(P<0.01),miR-20a-5p表达明显低于H8细胞(P<0.01)。宫颈癌组织中LncRNA HOTAIR和KIF26B表达明显升高(P<0.01),miR-20a-5p表达明显降低(P<0.01)。LncRNA HOTAIR、KIF26B表达降低后明显抑制了Hela细胞增殖与EMT,促进Hela细胞凋亡;LncRNA HOTAIR靶向miR-20a-5p,miR-20a-5p靶向KIF26B;miR-20a-5p表达降低后促进Hela细胞增殖与EMT,抑制细胞凋亡;过表达LncRNA HOTAIR通过miR-20a-5p促进Hela细胞增殖与EMT,抑制Hela细胞凋亡。结论LncRNA HOTAIR通过miR-20a-5p/KIF26B轴抑制Hela细胞凋亡,促进了癌细胞增殖及EMT。

Objective To investigate the effects of long chain noncoding RNA(LncRNA)HOTAIR on the proliferation,apoptosis and epithelial mesenchymal transformation(EMT)of cervical cancer Hela cells and its potential mechanism.Methods The cultured Hela and H8 cells,cervical cancer tissues and adjacent normal tissue samples were collected and the mRNA expressions of LncRNA HOTAIR,miR-20a-5p and KIF26B were detected by real-time quantitative PCR.The expression of LncRNA HOTAIR,miR-20a-5p and KIF26B were down-regulated,respectively,the proliferation ability of Hela cells was detected by MTT assay.The expressions of apoptosis related proteins Bax and Bcl-2 and EMT-related proteins E-cadherin and N-cadherin in Hela cells were detected by Western blot,and the apoptosis of Hela cells was detected by flow cytometry.The target and correlation between LncRNA HOTAIR and miR-20a-5p were analyzed by miRanda and dual luciferase reporter gene assay.The target and correlation between miR-20a-5p and KIF26B were analyzed by TargetScan and dual luciferase reporter gene assay.The effects of LncRNA HOTAIR on the proliferation,apoptosis and EMT of Hela cells via miR-20a-5p were detected.Results The expressions of LncRNA HOTAIR and KIF26B in Hela cells were significantly higher than those in H8 cells(P<0.01),the expression of miR-20a-5p was significantly lower than that of H8 cells(P<0.01).The expressions of LncRNA HOTAIR and KIF26B were significantly increased in cervical cancer tissues(P<0.01),the expression of miR-20a-5p was significantly decreased(P<0.01).The decreased expression of LncRNA HOTAIR and KIF26b significantly inhibited the proliferation and EMT of Hela cells,and promoted the apoptosis of Hela cells.LncRNA HOTAIR targeted miR-20a-5p and miR-20a-5p targeted KIF26B.Downregulation of miR-20a-5p expression promoted Hela cell proliferation and EMT,and inhibited cell apoptosis.Overexpression of LncRNA HOTAIR promoted the proliferation and EMT of Hela cells through miR-20a-5p,and inhibited the apoptosis of Hela cells.Conclusion LncRNA HOTAIR inhibited apoptosis of Hela cells through miR-20a-5p/KIF26B axis,and promoted cell proliferation and EMT.