RORγ基因对结肠癌细胞增殖及迁移的影响

Effect of RORγgene on proliferation and migration of colon cancer cells

目的 探究维甲酸相关孤儿核受体-γ(RORγ)基因对人结肠癌细胞增殖、转移能力的影响。方法 通过构建RORγ敲减细胞系,采用RT-qPCR及Western blot检测敲减效率;MTT、克隆形成、Transwell和划痕实验检测细胞增殖与转移能力;采用Western blot检测上皮间质转化(EMT)相关蛋白的表达情况;通过TIMER 2.0数据库分析RORγ基因的表达与肿瘤微环境中免疫细胞浸润的关系。结果 RORγ基因的敲减增强了结肠癌细胞的活力(F值=157.40,P<0.01)、克隆形成能力(F=61.35,P<0.01)、迁移能力(F=13.00,P<0.01)、侵袭能力(F=21.26,P<0.01)和划痕愈合能力(F=877.2,P<0.01),抑制了上皮型钙黏附蛋白(E-Cadherin)的表达,并促进了波形蛋白(vimentin)和神经型钙黏附蛋白(N-Cadherin)的表达;TIMER 2.0数据库分析显示结肠腺癌(COAD)组织中RORγ的表达水平与多种免疫细胞浸润相关。结论 RORγ表达的下调促进结肠癌细胞的增殖及转移。

Objective To investigate the effects of retinoid-related orphan receptorγ(RORγ)gene on proliferation and metastasis of human colon cancer cells.Methods RORγknockdown cell lines were constructed and the knockdown efficiency was detected by RT-qPCR and Western blot assays;MTT,colony formation,Transwell and wound healing assays were used to detect cell proliferation and metastasis;the expression of epithelial-mesenchymal transition(EMT)related proteins was detected by Western blot.The relationship between RORγgene expression and immune cell infiltration in tumor microenvironment was analyzed using TIMER 2.0 database.Results The knockdown of RORγenhanced the viability(F=157.40,P<0.01),clonogenesis(F=61.35,P<0.01),migration(F=13.00,P<0.01),invasion(F=21.26,P<0.01)and wound healing ability(F=877.2,P<0.01)of colon cancer cells,inhibited the expression of E-Cadherin,and promoted the expression of vimentin and N-Cadherin.TIMER 2.0 database analysis showed that RORγexpression in colon adenocarcinoma(COAD)tissues was associated with multiple immune cell infiltrates.Conclusion Downregulation of RORγexpression promoted the proliferation and metastasis of colon cancer cells.