白杨素对人肝癌HepG2细胞的诱导分化作用

摘要

Objective To investigate the effects of chrysin(ChR) on the induction of differentiation and apoptosis-promoting of HepG 2 human primary hepatocacinoma cells. Methods The HepG 2 cells were cultured in vitro, and then treated with ChR and all-trans retinotic Acid (RA), respectively, the alterations of nucleocytoplasm and tubulin arrangement after Gimsa staining and Coomassie brilliant blue staining were observed. The survival rate and the inhibitory rates of HepG 2 cells were determine by trypan blue counting method and MTT assay. The Alpha-fetoprotein(AFP) secretory amounts of the cells were detected by radioimmunoassay(RIA). The activities of alkaline phosphatase(ALP) andγ-glutamyltranspeptidase(γ-GT) were assayed by enzymatic reaction kit. The synthesis of tyrosine-α-ketoglutaric acid transaminase(TAT) in cells were investigated by Diamondstone spectrophotometry. Results After treatment with ChR or RA at 1.0~100μmol/L for 48 h, the proliferation of HepG 2 cells were inhibited significantly, compared with vehicle group (P<0.05 or P<0.01), the inhibitory potency of both ChR and RA on HepG 2 cells was equivalent and indicated in dose-dependent manner. After treatment with 10μmol/L ChR or RA for 48 h, HepG 2 cells disaggregated and grew to spindle-shape, their nuclei became smaller and the number of nucleolus were fewer. Furthermore, tubulin arrangement of cells tended to be more ordered and the tubulin synthesis increased significantly. At 24~96 hours treated with 10μmol/L ChR, the activities of TAT and ALP in cells were all increased distinctly (P<0.05, P<0.01), and the secretory amounts of AFP and the specific activities ofγ-GT were decreased significantly (P<0.05, P<0.01). Conclusion Chrysin can inhibit the proliferation of HepG 2 cells and induce them to differentiate to mature cells.%目的：探讨白杨素对人肝癌HepG2细胞的诱导分化和凋亡作用。方法体外培养人肝癌HepG2细胞，以全反式维甲酸为阳性对照，用白杨素干预细胞，台盼蓝计数法和MTT法检测药物对细胞增殖活力的影响；瑞氏-姬姆萨和考马斯亮蓝染色观察细胞核质比和微管微丝排列的变化；放射免疫法检测细胞甲胎蛋白（Alpha-fetoprotein，AFP）的分泌量；酶促反应试剂盒检测细胞中γ-谷胺酰转肽酶（γ-glutamyltranspeptidase，γ-GT）和碱性磷酸酶（alkaline phosephatase，ALP）的活性；Diamondstone分光光度法测定细胞中酪氨酸α-酮戊二酸转氨酶（tyrosine-α-ketoglutaric acid transaminase，TAT）的合成情况。结果1～100μmol/L白杨素和全反式维甲酸处理HepG2细胞48 h后，能显著抑制肝癌细胞的增殖（P＜0.05，P＜0.01），2种药物对细胞增殖的抑制效价相当并存在量效关系。10μmol/L药物作用48 h，细胞的形态和微管微丝排列由肿瘤细胞向成熟细胞分化；药物处理24～96 h后，细胞AFP的分泌量和γ-GT的活性明显降低，ALP和TAT的活性则显著升高（P＜0.05，P＜0.01）。结论白杨素具有抑制人肝癌HepG2细胞增殖并诱导其向正常细胞分化的作用。