三氯化镧对体外培养人肝癌细胞的抑制作用

目的：研究三氯化镧（LaCl₃）对人肝癌细胞株SMMC-7721生长的影响及其作用机理。方法：体外培养的人肝癌细胞SMMC-7721，给予LaCl₃（0.05、0.10、0.50及1.00 mmol?L^-1）培养不同时间（1、3、5和7 d），观察癌细胞生长曲线、集落抑制率、培 养上清中甲胎蛋白(AFP)的变化。采用流式细胞术检测SMMC-7721细胞周期变化。免疫组织化学检测培养 细胞CyclinD1、 PCNA、 CDK₄及P16的表达情况。结果: MTT法显示0.50及1.00 m mol?L^-1LaCl ₃对SMMC-7721生长具有明显的抑制作用, 且具有剂量和时间的依赖性; 0.10和0.50 mm ol?L^-1的LaCl₃对肝癌细胞集落的形成抑制率分别为51%和67%（P<0.05）,1.00 mmol?L^-1 LaCl ₃的集落形成抑制率达97％（P<0.01 ）；不同剂量LaCl₃　 作用SMMC-7721细胞3 d后，细胞周 期发生明显变化，表现为G₀/G₁期细胞百分数增加，与对照组比较差异有显著性（P<0.01）。0.10、0.50及1.00 mmol?L^-1 LaCl_{3均可抑制AFP的分泌，其中以1.00 mmol?L^-1 LaCl3组作用最显著（P<0.01 ）。免疫细胞化学结果显示，LaC l3促进P16蛋白 的表达，使Cyclin D1、PCNA和CDK4蛋白的表达下降。结论: LaCl3 对SMMC-7721细胞生长具有明显的抑制作用, 其机制可能与阻断细胞周期进程、降低细胞的恶性程度有关。}

Inhibitory effects of LaCl3 on hepatocellular carcinoma

Objective To observe the inhibitory effect of LaCl_3 on the growth of human hepatocellular carcinoma cell lines SMMC-7721 and its mechanism.Methods The cells were treated with different doses of LaCl_3 and the growth curve,colony inhibitory rates,the levels of AFP of cultured cells were detected at 1,3,5 and 7 days after they were dealed with LaCl_3.The changes of cell cycle were detected by flow cytometry and the expressions of cell cycle protein CyclinD_1,PCNA,CDK_4 and P16 were observed by immunocytochemistry staining.Results MTT test showed that SMMC-7721 cell proliterating activity was obviously inhibited by LaCl_3 in a time and dose related manner at concentrations of 0.50 and 1.00 mmol·L~(-1).The colony forming inhibitory rate of 0.10,0.50 and(1.00 mmol·L~(-1))LaCl_3 were 51%, 67% and 97% (P<0.05,P<0.01).Cell cycle progress of SMMC-7721 changed obviously when treated with different does of LaCl_3,the percent of G_0/G_1 phase was higher and S phase lower,the difference was significant compared with control group(P<0.01).0.10,0.50 and 1.00 mmol·L~(-1) LaCl_3 could decreased the levels of AFP,the effect of 1.00 mmol·L~(-1) LaCl_3 was notable(P<0.01).Immunocytochemistry showed that LaCl_3 could enhance the expression of P16 protein and decrease the expressions of CyclinD_1,PCNA and CDK_4.Conclusion LaCl_3 could inhabit the growth of SMMC-7721 cell and its mechanism might be connected with the inhibition of cell cycle progress and decrease the degree of malignancy.