染料木黄酮对小鼠肝癌高淋巴转移细胞株HepA-H的抑制作用

THE INHIBITORY EFFECTS OF GENISTEIN ON MURINE HEPATOCARCINOMA CELL LINE (HepA-H) WITH HIGH LYMPHATIC METASTATIC ABILITY

目的:研究染料木黄酮(genistein,Gen)对腹水型小鼠肝癌高淋巴转移细胞株HepA-H的抑制作用及其机制。方法:MTT法检测Gen对HepA-H的增殖抑制作用。应用电镜、琼脂糖凝胶电泳及流式细胞术检测Gen诱导的HepA-H细胞凋亡。同时,将HepA-H皮下接种于NIH小鼠的足垫,Gen混悬于2%的卵磷脂中,从细胞接种的D2起,每日以100或200mg/(kgbw.d)腹腔给药,D15取淋巴结观察,并计算转移率。TUNEL法检测淋巴结内凋亡的HepA-H细胞,并计算凋亡指数。结果:Gen在体外对HepA-H细胞具有良好的增殖抑制作用,呈量效、时效关系,并能诱导HepA-H细胞凋亡;在体内能减少荷瘤动物肿瘤体积和抑制肿瘤淋巴转移,给药组凋亡指数明显高于对照组(P<0.01)。结论:Gen能明显抑制HepA-H肝癌细胞增殖和淋巴转移,其作用机制可能与诱导细胞凋亡有关。

Objective： To investigate the inhibitory effects of genistein （Gen） on murine hepatocarcinoma cell line with high lymphatic metastasis （HepA-H）, and explore its mechanism. Method： The inhibitory effect on the proliferation of HepA-H cells was evaluated by MTT assay. Gen-induced apoptosis in HepA-H cells was observed by scanning electron microscopy （SEM）, transmission electron microscopy （TEM）, DNA agarose electrophoresis and flow cytometry. HepA-H cells were injected subcutaneously to the foot pad of NIH mice. Gen was suspended in 2% lecithin and administered at the dose of 100 or 200 mg/（kg bw .d） by intraperitoneal injection daily from the day before cell injection. The popliteal lymph nodes were taken and the metastatic rates were calculated after 15 d. Using TUNEL method, the apoptosis of HepA-H cells in lymph nodes was checked and apoptosis index （AI） was calculated. Results： Gen exhibited remarkable antiproliferative activity in dose and time dependent manner, and induced apoptosis of HepA-H. Gen could reduce the tumor size and inhibit the metastasis to lymphtic nodes. AI of two Gen groups was higher than the control（P〈0.01）. Conclusion： Gen can inhibit proliferation of tumor cells and matastasis to lymphtic nodes through induction of tumor cell apoptosis. [ACTA NUTRIMENTA SINICA, 2006, 28（1）： 66-70]