摘要
目的探讨长链非编码RNA(lncRNA)HCG11对宫颈癌细胞增殖、凋亡、侵袭和迁移的影响及作用机制。方法采用实时荧光定量聚合酶链反应(qRT-PCR)法检测正常子宫内膜上皮细胞ESC以及宫颈癌细胞HeLa和Siha细胞中HCG11的表达水平。转染pcDNA3.1-HCG11或anti-miRNA-421至HeLa细胞,构建过表达HCG11或抑制miRNA-421表达的HeLa细胞,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell法检测细胞迁移和侵袭,Western blot检测细胞周期蛋白D1(cyclin D1)、P21、B细胞淋巴瘤/白血病-2(Bcl-2)、B细胞淋巴瘤-2相关X蛋白(BAX)、基质金属蛋白酶2(MMP2)和上皮钙黏素(E-cadherin)蛋白表达水平。结果与ESC细胞相比,HeLa和Siha细胞中HCG11表达水平明显降低(P﹤0.01)。过表达HCG11或抑制miRNA-421表达可使HeLa细胞光密度(OD)值明显降低,凋亡率明显升高,迁移和侵袭数明显减少,cyclin D1、Bcl-2和MMP2蛋白表达降低,P21、BAX和E-cadherin蛋白表达升高(P﹤0.05)。miRNA-421过表达逆转了过表达HCG11对HeLa细胞增殖、凋亡、迁移和侵袭及相关蛋白表达的影响。结论HCG11靶向负调控miRNA-421表达,抑制宫颈癌细胞增殖、侵袭和迁移,并诱导其凋亡。
Objective To investigate the effects of long non-coding RNA(lncRNA)HCG11 on the proliferation,apoptosis,invasion and migration of cervical cancer cells and its mechanism.Method The normal endometrial epithelial cells ESC and cervical cancer cells HeLa and Siha were cultured,in which the expression levels of HCG11 were detected by quantitative real-time polymerase chain reaction(qRT-PCR).After transfecting with pcDNA3.1-HCG11 or anti-miRNA-421 into HeLa cells,HCG11 overexpression or inhibited miRNA-421 expression were established,afterwards,cell proliferation was detected by MTT,apoptosis was determined by flow cytometry,and cell migration and invasion were analyzed using Transwell assay,the expression of cyclin D1,P21,B cell lymphoma/leukemia-2(Bcl-2),Bcl-2-associated X protein(BAX),matrix metalloproteinase 2(MMP2)and E-cadherin were detected by Western blot.Result Compared with ESC cells,the expression of HCG11 in HeLa and Siha cells were significantly lower(P<0.01).Overexpression of HCG11 or inhibited miRNA-421 expression significantly reduced the optical density(OD)value of HeLa cells,while increased the apoptosis rate,and reduced migration and invasion,besides,the expression of cyclin D1,Bcl-2 and MMP2 proteins were markedly decreased,though P21,BAX and E-cadherin proteins expression evidently elevated(P<0.05).Finally,miRNA-421 overexpression reversed the effect induced by HCG11 overexpression on proliferation,apoptosis,migration and invasion,and related protein expression of HeLa cells.Conclusion HCG11 may inhibit the proliferation,invasion and migration and induce apoptosis of cervical cancer cells by negatively regulating miRNA-421 expression.
作者
刘秀玲
陈新宇
王志红
李静
贺全勤
LIU Xiuling;CHEN Xinyu;WANG Zhihong;LI Jing;HE Quanqin(Department of Gynecology,Zhumadian Central Hospital,Zhumadian 463000,He’nan,China;Department of Oncology,Zhumadian Central Hospital,Zhumadian 463000,He’nan,China;Department of Obstetrics and Gynecology,Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,He’nan,China;Department of Obstetrics and Gynecology,Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,He’nan,China)
出处
《癌症进展》
2020年第1期30-34,38,共6页
Oncology Progress
