CXCL-1基因稳定转染肝癌细胞株的建立、鉴定及其细胞生物学特性

Establishment, identificaton and biological features of stably CXCL1 transfected human hepatocellular carcinoma cells

目的 建立慢病毒载体介导稳定转染趋化因子CXCL-1的MHCC97-L肝癌细胞株,鉴定转染细胞中目的基因的表达并观察转染CXCL-1对MHCC97-L细胞增殖、迁移、侵袭能力及细胞周期的影响.方法 慢病毒载体介导CXCL-1基因稳定转染MHCC97-L肝癌细胞,qRT-PCR、蛋白印迹方法分别检测转染后细胞中CXCL-1在mRNA和蛋白水平的表达;MTT试验、克隆形成试验,Transwell小板细胞穿膜试验分别检测转染后细胞增殖能力、迁移能力及侵袭能力的变化.结果 成功建立了稳定转染CXCL-1的MHCC97-L肝癌细胞株.转染后细胞高表达CXCL-1基因.转染后MHCC97-L细胞增殖能力显著增强（P＜0.05）,侵袭能力显著增强（P＜0.05）,迁移能力变化则无统计学差异（P＞0.05）.转染后细胞株中G1期细胞比例显著减少（P＜0.05）.结论 成功构建稳定转染CXCL-1的肝癌细胞株,CXCL-1可增强肝癌细胞的体外增殖和侵袭能力.

Objective To establish a MHCC97-L cell lines which stably express CXCL-1 by using lentiviral transfection system and investigate the effect of CXCL1 on the cell proliferation,migration and invasion as well as the cell cycle.Methods The expression levels of mRNA and protein in MHCC97-L cell line with the CXCL1 lentivirus-mediated transfection were evaluated by qRT-PCR and western blot,respectively.We conducted MTT,clone formation and Transwell assay to examine the changes of cell proliferation,migration and invasion in the transfected cells.Results A stably transfected cell line was successfully established.Cell proliferation （P ＜ 0.05） and invasion （P ＜ 0.05） of MHCC97-L cells transfected with CXCL-1 gene tended to be significantly enhanced whilst nothing seemed to alter in cell migration （P ＞ 0.05）.The percentage of transfected cells in the G1 stage declined significantly （P ＜ 0.05）.Conclusion A stable cell line transfected with CXCL1 was established,and CXCL1 overexpression could promote the proliferation and invasion of the hepatocellular carcinoma cells in vitro.