金雀异黄素对HepG2细胞的生长抑制与诱导分化作用

Induction of Proliferation Inhibition and Differentiation of Hepatocarcinoma Cell HepG2 by Genistein

关键词:  金雀异黄素  人肝癌HepG2细胞  生长抑制  细胞分化

Key words:  genistein  liver cancer cell  proliferation inhibition  cell differentiation

摘要:

目的 观察黄酮金雀异黄素(genistein，Gen)抑制人肝癌HepG2细胞的增殖和诱导其分化的作用。方法 体外培养HepG2细胞，以全反式维甲酸(RA)为阳性对照，MTT比色法测定Gen对HepG2细胞增殖的影响；细胞计数法测定HepG2细胞存活率；瑞氏-姬姆萨染色法和考马斯亮蓝染色法分别观察Gen对HepG2细胞形态、细胞微管蛋白排列的影响；放免法测定HepG2细胞AFP的分泌量；酶促反应试剂盒检测细胞中γ-GT、ALP；Diamondstone分光光度法测定细胞中TAT的含量。结果 MTT法和细胞计数法检测结果都表明，不同浓度(10 μmol·L^-1、100 μmol·L^-1)的Gen诱导人肝癌HepG2细胞48 h，细胞增殖活力明显降低，并呈剂量依赖性下降。10 μmol·L^-1 Gen孵育HepG2细胞48 h后细胞形状由圆形转变为长梭形、细胞相互分散、细胞核变小、核仁的数目减少；微管微丝增多，排列更整齐，细胞趋向成熟分化。以10 μmol·L^-1 Gen分别处理HepG2细胞24，48，72和96 h后，显著地降低了γ-GT的活性和AFP的表达，增加了TAT和ALP的活性。结论 Gen能抑制人肝癌HepG2细胞的生长和增殖，并能诱导其向成熟细胞分化。

Abstract:

OBJECTIVE To investigate the effects of genistein on proliferation inhibition and differentiation induction of HepG2 human primary hepatocacinoma cells. METHODS The HepG2 cells were treated with genistein or all-trans retinotic acid to observe the alterations of nucleocytoplasm and tubulin arrangement after Gimsa coloration and Coomassie brilliant blue staining. MTT and cell counting method were used to detect the effect of genistein on the proliferation of HepG2 cells. the Alpha-fetoprotein(AFP) secretary amount of the cells was assayed by radioimmunoassay(RIA), the activities of alkaline phosephatase(ALP) and γ-glutamyl transpeptidase(γ-GT) were investigated by enzymatic reaction kit, the synthesis of TAT was determined by Diamondstone spectrophotometry. RESULTS After treated with 10 μmol·L^-1 and 100 μmol·L^-1 genistein for 48?h respectively, the proliferation of HepG2 cells were all obviously inhibited, and the inhibition degrees were related to dose of the drugs(P<0.05, P<0.01). Treated with 10 μmol·L^-1 genistein, the nucleocytoplasm ratio of the cells was reduced significantly (P<0.01), the specific activities of TAT and ALP, biomarkers of liver cancer cells, were all increased evidently, and the secretory amount of AFP and the specific activities of γ-GT were decreased obviously(P<0.01). CONCLUSION Genistein can inhibit the proliferation of HepG2 human primary hepatocacinoma cells, and induce them to differentiate to mature cells.