TMEM9B-AS1对食管癌细胞增殖和侵袭影响及机制

EFFECT OF TMEM9B-AS1 ON THE PROLIFERATION AND INVASION OF ESOPHAGEAL CANCER CELLS AND RELATED MECHANISM

目的探讨长链非编码RNA(LncRNA)TMEM9B-AS1介导TCF7L2-SATB1途径调控食管癌细胞环氧化酶-2(COX-2)和β连环蛋白1(CTNNB1)表达以及对细胞增殖和侵袭的影响。方法将人食管癌细胞KYSE510分为对照组、TMEM9B-AS1组和siTMEM9B-AS1组,分别转染阴性对照(NC)质粒、TMEM9B-AS1质粒和siTMEM9B-AS1质粒。通过细胞计数(CCK-8)法和转移小室(Transwell)检测细胞增殖活力和侵袭能力。通过定量逆转录聚合酶链式反应(RT-qPCR)和蛋白质印迹(Westernblot)检测转录因子7样蛋白2(TCF7L2)、特殊富含AT的序列结合蛋白1(SATB1)、COX-2、CTNNB1的mRNA和蛋白表达水平。结果与对照组相比较,TMEM9B-AS1组细胞的TMEM9B-AS1水平、侵袭能力以及TCF7L2、SATB1、COX-2、CTNNB1的mRNA和蛋白水平显著升高(F=30.268~56.084,P<0.05),siTMEM9B-AS1组细胞则显著降低(P<0.05)。各组细胞中增殖活力对比,差异具有统计学意义(F_(组别)=18.681,P<0.001);随着培养时间的延长,各组细胞的增殖活力显著上调(F_(时间)=4.347,P<0.001)。结论过表达TMEM9B-AS1可促进TCF7L2-SATB1途径,提高COX-2和CTNNB1的表达,并促进食管癌细胞的增殖和侵袭的能力。

Objective To investigate the regulatory effect of the long non-coding RNA(LncRNA)TMEM9B-AS1 on the expression of cyclooxygenase-2(COX-2)and beta-catenin 1(CTNNB1)in esophageal cancer cells by mediating the TCF7L2-SATB1 pathway and its effect on the proliferation and invasion of esophageal cancer cells.Methods Human esophageal cancer KYSE510 cells were divided into control group,TMEM9B-AS1 group,and siTMEM9B-AS1 group and were transfected with negative control plasmid,TMEM9B-AS1 plasmid,and siTMEM9B-AS1 plasmid,respectively.CCK-8 assay and Transwell assay were used to measure cell proliferation activity and invasion ability.Reverse transcription-quantitative PCR and Western blot were used to measure the mRNA and protein expression levels of transcription factor 7-like protein 2(TCF7L2),special AT-rich sequence-binding protein 1(SATB1),COX-2,and CTNNB1.Results Compared with the control group,the TMEM9B-AS1 group had significant increases in the level of TMEM9B-AS1,the invasive ability of cells,and the mRNA and protein expression levels of TCF7L2,SATB1,COX-2,and CTNNB1(F=30.268-56.084,P<0.05),while the siTMEM9B-AS1 group had significant reductions in these indices(P<0.05).There was a significant difference in cell proliferation activity between groups(F_(group)=18.681,P<0.001),and cell proliferation activity significantly increased over the culture time(F_(time)=4.347,P<0.001).Conclusion Overexpression of TMEM9B-AS1 can promote the TCF7L2-SATB1 pathway,increase the expression of COX-2 and CTNNB1,and promote the proliferation and invasion abilities of esophageal cancer cells.