摘要
目的探索核仁小分子RNA宿主基因6(SNHG6)在结肠癌组织中的表达特点,以及其对结肠癌细胞增殖、迁移和侵袭能力的影响。方法采用荧光定量PCR检测25例患者结肠癌组织及对应癌旁组织中SNHG6mRNA的表达水平。将LoVo细胞分成Blank组、NC组、siRNA-1组和siRNA-2组,其中Blank组不作任何处理,另外3组依次使用Lipofectamine 2000转染NC-siRNA、SNHG6-siRNA-1及SNHG6-siRNA-2;通过CCK-8实验测定细胞增殖能力,通过Transwell实验测定细胞迁移及侵袭能力;通过荧光定量PCR和蛋白质印迹法(Western blot)检测上皮-间质转化(EMT)标志物上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、Snail的mRNA及蛋白表达水平。结果患者结肠癌组织SNHG6mRNA表达水平高于对应癌旁组织(U=138.0,P=0.000 7)。siRNA-1组和siRNA-2组细胞增殖活性较Blank组明显下降(P<0.05);siRNA-1组和siRNA-2组细胞的迁移和侵袭数量较Blank组明显减少(P<0.05);siRNA-1组、siRNA-2组与Blank组比较,E-Cadherin mRAN及蛋白表达上调,N-Cadherin、Vimentin及Snail mRNA及蛋白表达下调,差异有统计学意义(P<0.05)。结论 SNHG6在结肠癌组织中高表达,敲低SNHG6表达能够抑制结肠癌细胞EMT进程,进而抑制细胞的增殖、迁移及侵袭能力。
Objective To investigate the expression characteristics of small-nucleolar-RNA host gene 6（SNHG6）in colon cancer tissues,and explore its biological function on colon cancer cell lines LoVo,including the cell proliferation,migration and invasion.Methods The expression levels of SNHG6 mRNA in colon cancer tissues and corresponding adjacent normal tissues obtained from 25 patients were detected by real-time polymerase chain reaction（qRT-PCR）.The LoVo cells were divided into four groups,including the blank group,the negative control（NC）group,the siRNA-1 group and the siRNA-2 group.Using lipofectamine 2000,the NC group,the siRNA-1 group and the siRNA-2 group were respectively transfected with NC-siRNA,SNHG6-siRNA-1 and SNHG6-siRNA-2,while the blank group was not transfected.The ability of cell proliferation was measured by cell counting kit-8（CCK-8）.The abilities of cell migration and invasion were measured by transwell.The expression levels of mRNA and protein of epithelial-mesenchymal transition（EMT）markers,including E-cadherin,N-cadherin,Vimentin and Snail,were detected by both qRT-PCR and western blot.Results The expression level of SNHG6 mRNA was higher in colon cancer tissues than that in the the corresponding adjacent normal tissues（U=138.0,P=0.000 7）.LoVo cells in the siRNA-1 group and the siRNA-2 group were suppressed when compared with the blank group（P〈0.05）.The number of cells migrated and invaded in the siRNA-1 group and siRNA-2 group were significantly smaller than those in the blank group（P〈0.05）.The expression levels of E-cadherin mRNA and protein in the siRNA-1 group and the siRNA-2 group were significantly up-regulated,while N-cadherin,Vimentin and Snail mRNA and protein in the siRNA-1 group and the siRNA-2 group were down-regulated,compared with the blank group,there were statistically significant differences（P〈0.05）.Conclusion SNHG6 is highly expressed in colon cancer tissues.Down-regulating the expression of SNHG6 could inhibit progression of EMT,and inhibit cell proliferation,migration and invasion.
作者
黎丰华
古国财
庄雄标
吴印爱
刘献棠
王志伟
LI Fenghua;GU Guocai;ZHUANG Xiongbiao;WU Yin′ai;LIU Xiantang;WANG Zhiwei(Department of General Surgery,157 Branch Hospital,General Hospital of GuangzhouMilitary Command of PLA,Guangzhou,Guangdong 510515,China)
出处
《重庆医学》
CAS
2018年第18期2413-2417,共5页
Chongqing medicine
关键词
结肠肿瘤
核仁小分子RNA宿主基因6
上皮-间质转化
细胞增殖
细胞迁移
侵袭
colonic neoplasms；small-nucleolar-RNA host gene 6；epithelial-mesenchymal transition；cell proliferation；cell migration；invasiveness
