敲低TROAP基因对胆管癌细胞系增殖和迁移能力的影响

Effects of TROAP Gene Knockdown on the Proliferation and Migration of Cholangiocarcinoma Cell Lines

目的:探究Trophinin相关蛋白(Trophinin-associated protein,TROAP)基因沉默对人胆管癌细胞系RBE细胞增殖和迁移能力的影响。方法:培养人胆管癌细胞系RBE,并对其分别转染阴性对照序列和TROAP干扰序列,分别命名为si-NC组和siTROAP组。采用细胞计数试剂盒(CCK-8)和平板克隆形成实验检测细胞增殖能力;流式细胞术检测细胞凋亡率;细胞划痕实验和Transwell法检测细胞迁移和侵袭能力;蛋白质印迹实验(Western blot)检测RBE细胞中上皮间充质化(EMT)相关蛋白E-cadherin、N-cadherin,PCNA以及凋亡相关蛋白Bcl-2、Bax和caspase-3的相对表达量。结果:与si-NC组细胞相比,si-TROAP组细胞增殖率降低,克隆个数降低,细胞凋亡率升高,迁移和侵袭能力降低(P<0.05)。与si-NC组细胞相比,si-TROAP组细胞E-cadherin蛋白相对表达水平升高,N-cadherin蛋白相对表达水平降低,Bax和caspase-3蛋白相对表达水平升高,Bcl-2和PCNA蛋白相对表达水平降低(P<0.05)。结论:TROAP基因是RBE细胞潜在的致癌基因,TROAP基因沉默可降低人胆管癌细胞系RBE细胞增殖和迁移能力,并通过缓解上皮间质转化来发挥效应。

Objective:To investigate the effects of Trophinin-associated protein(TROAP)gene silencing on the proliferation and migration of human cholangiocarcinoma cell line RBE.Methods:RBE cells were cultured and transfected with negative control sequence and TROAP interference sequence in si-NC group and si-TROAP group respectively.Cell proliferation ability was detected by cell counting kit-8(CCK-8)and plate clone formation experiment.Apoptosis rate was detected by flow cytometry.Cell migration and invasion ability were detected by cell scratch test and Transwell method.The relative expressions of epithelial-mesenchymal transition(EMT)related proteins E-cadherin,N-cadherin,PCNA and apoptosis related proteins Bcl-2,Bax and caspase-3 in RBE cells were detected by Western bloting.Results:Compared with si-NC group,the cell proliferation rate and number of clones decreased,the cell apoptosis rate increased,and the cell migration and invasion ability decreased in the si-TROAP group(P<0.05).Compared with si-NC group,the relative expression levels of E-cadherin protein increased,and the relative expression levels of N-cadherin protein decreased in the si-TROAP group;the relative expression levels of Bax and caspase-3 increased,and the relative expression levels of Bcl-2 and PCNA protein decreased in the si-TROAP group(P<0.05).Conclusion:TROAP gene is a potential oncogene in RBE cells.TROAP silencing can reduce the proliferation and migration ability of human cholangiocarcinoma cell line RBE by alleviating epithelial-mesenchymal transition.