敲低Ro60/SSA表达对肝癌细胞侵袭和转移能力的影响

Effects of down-regulating Ro60/SSA expression on migration and invasion ability of hepatocellular carcinoma cells

目的:探讨下调Ro60/SSA的表达对肝癌细胞侵袭和转移能力的影响。方法:将人肝癌细胞HAK-1A和HepG2分别分3组处理:空白对照组常规培养(DMEM培养基+体积分数10%胎牛血清);另两组脂质体法分别转染siRNA-NC和siRNA-Ro60/SSA,每组设置3个复孔。转染48 h后应用Western blot法检测细胞中Ro60/SSA蛋白的表达,应用克隆形成实验和球形形成实验检测细胞的自我更新和增殖能力;应用Transwell小室法检测3组HAK-1A细胞的迁移和侵袭能力。将雌性BALB/c裸鼠随机分为siRNA-NC组和siRNA-Ro60/SSA组,每组5只。将转染siRNA-NC或siRNA-Ro60/SSA的HAK-1A细胞以1×10^(4)个/μL尾静脉注射入裸鼠体内(200μL/只),4周后,计数肺部肿瘤灶数量。结果:与空白对照组和siRNA-NC组比较,siRNA-Ro60/SSA组HAK-1A和HepG2细胞Ro60/SSA蛋白表达下调,克隆形成数、成球率均降低(P<0.05)。siRNA-Ro60/SSA组HAK-1A细胞Transwell实验中的迁移细胞数和侵袭细胞数低于空白对照组和siRNA-NC组(P<0.05)。siRNA-Ro60/SSA组裸鼠肺部移植瘤数量低于siRNA-NC组(P<0.05)。结论:敲低Ro60/SSA可明显抑制肝癌细胞的侵袭和转移能力。

Aim:To investigate the effects of down-regulating Ro60/SSA expression on migration and invasion ability of hepatocellular carcinoma cells.Methods:Hepatocellular carcinoma HAK-1A and HepG2 cells were allocated into blank control(cultured with DMEM and 10%fetal bovine serum),siRNA-NC(transfected with siRNA-NC for 48 hours)and siRNA-Ro60/SSA(transfected with siRNA-Ro60/SSA for 48 hours)groups.Western blot was used to detect the expression of Ro60/SSA protein in the cells,spheroid and colony formation assays were used to analyze self-renewal and proliferative ability.Transwell method was performed to detect the migration and invasion ability of HAK-1A cells from the 3 groups.HAK-1A cells transfected with siRNA-NC or siRNA-Ro60/SSA were injected into nude mice by 1×10^(4) cells/μL through tailveins(200μL per nude mouse),after 4 weeks,the number of tumor foci in the lung was counted,5 nude mice in each group.Results:Compared with those of blank control group and siRNA-NC group,Ro60/SSA expression in HAK-1A and HepG2 cells from siRNA-Ro60/SSA group was down-regulated,and the spheroid rate and colony number decreased(P<0.05).The number of migration and invasion HAK-1A cells from siRNA-Ro60/SSA group was lower compared with the blank control group and siRNA-NC group(P<0.05).The transplanted tumor number in the lung of nude mice from siRNA-Ro60/SSA group was lower than that of siRNA-NC group(P<0.05).Conclusion:Knockdown the expression of Ro60/SSA may suppress the migration and invasion ability of hepatocellular carcinoma cells.