肝细胞生长因子体外抑制SMMC-7721人肝细胞癌细胞生长

摘要

目的:观察肝细胞生长因子(HGF)对体外培养的人肝细胞癌细胞的生长效应.方法:将处于指数生长期的SMMC-7721细胞经过(HGF处理组)或不经(对照组)HGF处理培养1～4d,采用四甲基偶氮唑盐(MTT)法检测细胞数,并进行细胞周期分析.结果:HGF对SMMC-7721细胞增殖具有剂量依赖性抑制作用.在实验浓度内,使用最高浓度的HGF(rhHGF为20μg*L-1、cHGF为100mg*L-1)对细胞增殖抑制作用最强.使用不同浓度(5、10、20μg*L-1)rhHGF处理3d的培养细胞均有一半以上停留在G0/G1期.结论:HGF对SMMC-7721人肝细胞癌细胞的体外生长有强烈的抑制作用,这与细胞生长阻滞在G0/G1期有关.%Objective To make sure whether hepatocyte growth factor(HGF) is a negative growth regulator for human hepatocellular carcinoma(HCC) cells or not,we observed the growth effects of both recombinant human HGF(rhHGF) and native calf HGF(cHGF) on SMMC-7721 human HCC cell line,which was derived from the hepatocarcinoma tissue of a Chinese HCC patient.Methods SMMC-7721 human HCC cells were cultured to the exponential phase of growth and then treated with(HGF-treated group)or without(control) rhHGF/cHGF.After 1～4 d,the number of viable cells was determined by a photometric method using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT). The cells incubated with rhHGF were subjected to analysis of cell cycle distribution by flow cytometric assay.Results A similar type of dose-dependent cell growth inhibition effect on SMMC-7721 human HCC cells by rhHGF(5～20μg*L-1) as well as by cHGF(25～100mg*L-1) has been found, with the maximal effect seen at the highest concentration used. Approximately over 50% of the cells treated with rhHGF(5μg*L-1,10μg*L-1,20μg*L-1) accumulated in the quiescent G0/G1 phase of the cell cycle over incubation period for 3d.Conclusion The growth of SMMC-7721 human HCC cells is strongly inhibited by rhHGF or cHGF.The cells exposed to HGF are arrested in the G0/G1 phase of the cell cycle.Thus,our study prove that HGF may be a negative growth regulator for human HCC cells.