LHRH类似物对体外培养肝癌细胞增殖阻抑作用的实验研究

Experimental study on the inhibition of hepatocarcinoma cell proliferation by luteinizing hormone-releasing hormone analog

目的 :探讨黄体素释放激素 (LHRH)类似物对肝癌细胞增殖的调节作用 ,为阐明LHRH在肝癌发病中的作用机制提供实验资料。 方法 :以体外培养的人肝癌细胞系hHCC和大鼠肝癌细胞系FSK 790 2为研究对象 ,用MTT试验检测LHRH类似物对这两种肝癌细胞生长的抑制作用 ;用流式细胞仪测定LHRH类似物作用后癌细胞的细胞周期变化 ;用免疫组化染色法检测PCNA和Bax蛋白表达以反映LHRH类似物对癌细胞增殖和程序性死亡指标的影响。 结果 :MTT试验表明 ,LHRH类似物可抑制hHCC和FSK 790 2的生长 ,并且这种作用呈时间和剂量依赖关系。LHRH类似物作用后 ,肝癌细胞中G1/G0 期细胞比率上升 ,S期细胞比率下降 ,说明细胞生长停滞。LHRH类似物可使癌细胞中PCNA免疫反应阳性细胞比率下降 (P <0 .0 5 ) ;而Bax阳性细胞比率增加 (P <0 .0 5 )。 结论 :LHRH类似物具有体外抑制肝癌细胞增殖的活性 ;

Objectives:To investigate the regulating effects of luteinizing hormone releasing hormone analog(LHRH A) on hepatocellular carcinoma(HCC) cell proliferation and to elucidate the mechanism of LHRH in HCC development. Methods:The in vitro antitumor effects of LHRH A on human and rat hepatocarcinoma cell lines, hHCC and FSK 7902,were detected by using microculture tetrazolium cytotoxicity test(MTT) for cell growth, flow cytometry(FCM) for cell cycle analysis and by immunocytochemistry staining for cell marks of proliferation and apoptosis. Results:MTT test showed that LHRH A inhibited hHCC and FSK 7902 cell proliferating activity in a time and dose related manner at concentrations ranging from 100 nmol/L to 100 μmol/L(exposure lasting 48 to 96 hours).Cell cycle phase DNA analysis by flow cytometry demonstrated that LHRH A caused a significant increase in the portion of cells in the G 1/G 0 phase and a marked decrease in the portion of cells in S phase, which indicating cell growth retardation. After LHRH A treatment, the PCNA positive cell percentage decreased and Bax positive cell percentage increased in both cell lines( P <0.05). Conclusions: LHRH A is effective in inhibiting the hepatocarcinoma cell proliferation, and Bax might be involved in the inhibitory effect of LHRH A on hepatocarcinoma cells.