中药复方胃肠安血清诱导肝癌SMMC-7721细胞分化  

Differentiation of hepatocellular carcinoma SMMC-7721 cell line induced by Chinese medicine recipe Weichangan

作　　者：赵海磊[1] 刘成[2] 赵爱光[1] 

机构地区：[1]上海中医药大学附属龙华医院肿瘤一科,上海市200032 [2]上海中医药大学肝病研究所,上海市200032

出　　处：《世界华人消化杂志》2003年第9期1345-1348,共4页World Chinese Journal of Digestology

基　　金：上海市青年科技启明星计划;No.02QB14043;上海市教委基金;No.01C06

摘　　要：目的:观察中药复方胃肠安诱导肝癌细胞分化的作用.方法:以SMMC-7721人肝癌细胞为研究对象,维甲酸为对照,采用药物血清添加法,通过 MTT法和 Alamar Blue法观察胃肠安对肝癌细胞增生的抑制作用;放射免疫法观察对肝癌细胞分泌甲胎蛋白和白蛋白的影响;Western blot观察对肝癌细胞P53、P16以及P21蛋白表达的影响.结果:MTT法显示胃肠安与维甲酸对SMMC-7721人肝癌细胞增生均有抑制作用,胃肠安在48 h达到最大,而维甲酸对细胞增生的抑制作用在48 h后逐渐下降,较胃肠安组明显降低.Alamar Blue法结果显示,16 h后,胃肠安组细胞还原的 Alamar Blue值较对照组明显减少,并在32 h后较维甲酸组显著下降,提示与维甲酸相比,胃肠安作用起效慢但持续时间长;胃肠安组分泌的甲胎蛋白较对照组显著减少(11.4 ±1.4μg/L vs 17.2 ± 1.1μg/L,P=0.036),而白蛋白显著增多(0.40 ± 0.02mg/L vs 0.29 ± 0.01mg/L,P=0.043);胃肠安组突变型P53蛋白的表达较对照组明显减少,而P16蛋白和P21蛋白的表达较对照组增多.结论:胃肠安方剂具有抑制SMMC-7721人肝癌细胞增生、诱导细胞分化的作用,其机制可能在于减少突变型P53蛋白表达和增加P16,P21蛋白表达。AIM: To observe the differentiation in hepatocellular carci- noma cell line induced by Chinese medicine recipe Weichangan. METHODS: Weichangan, contrasted by the retinoic acid and distilled water, was made by using serum pharmacological method. The inhibition on the growth of SMMC-7721 cell line by Weichangan was observed through the method of MTT and Alamar Blue. Radioimmunoassay was applied to determine the concentration of α-fetoprotein and albumin secreted by the incubated cells. Western blot method was used to detect the mutant p53, p16 and p21 protein expression in SMMC-7721 cell line. RESULTS: MTT assay showed both Weichangan and serum retinoic acid had inhibiting effect on the proliferation of human hepatocellular carcinoma SMMC-7721 cell line. Weichangan reached its maximal inhibition effect after 48 hours, while the effect of retinoic acid decreased gradually after 48 hours. Alamar Blue method showed that significant decrease was found in serum Weihangan after 16 hours compared with that in the control. After 32 hours, the decrease induced by Alamar Blue was more significant than that in cells incubated with serum Weichangan compared with those incubated in serum retinoic acid, indicating the gradual and durable action of Weichangan recipe. The decreased amount (11.4±1.4 μg/L vs 17.2±1.1 μg/L, P=0.036) of α- fetoprotein and increased amount (0.40±0.02 mg/L vs 0.29±0.01 mg/L, P=0.043) of albumin were found in the cells incubated in serum Weichangan. Western blot method showed decreased expression of p53 protein and increased expression of p16 and p21 protein in cells incubated in serum Weichangan. CONCLUSION: The results suggest that Weichangan inhibits the growth of SMMC-7721 cell line and induces the differen- tiation in this hepatocellular carcinoma cell line. The effect on p53, p16 and p21 may be the mechanisms of Weichangan in inducing the differentiation of this cell lines.

关 键 词：中医药疗法 胃肠安 血清 肝癌 SMMC—7721细胞 细胞分化 甲胎蛋白 白蛋白 放射免疫法 

分 类 号：R735.7[医药卫生—肿瘤]