目的 探讨大豆甙元(daidzein)诱导H22肿瘤细胞凋亡的作用及机制。方法用四甲基偶氮噻唑蓝(MTT)法检测不同剂量大豆甙元(0.1～150.0μmol/L)对H22细胞增殖的影响;昆明雄性小鼠60只,随机分为6组,每组10只,腹腔注射给药,分别为模型组、对照组(不含血清的RPMI 1640培养液),低、中、高剂量大豆甙元组(大豆甙元分别为501、00、200 mg/kg),阳性对照组(环磷酰胺30 mg/kg);给药14 d后比较各组小鼠体重、脾脏指数、胸腺指数及瘤重;采用western blot检测bax、bcl-2的蛋白表达。结果 MTT检测结果显示大豆甙元对H22细胞有抑制作用,150μmol/L大豆甙元作用72 h,抑制率达30.3%,与低剂量组比较有明显抑制作用(P<0.05);大豆甙元能使Bax表达上调,Bcl-2表达下调,与对照组比较,10.0μmol/L大豆甙元组bax表达灰度值增加50.05%,凋亡抑制基因bcl-2表达减少52.62%;中、高剂量大豆甙元组小鼠胸腺指数和脾脏指数均明显升高(P<0.01),与模型组比较,大豆甙元各组瘤重指数下降明显(P<0.01)。结论 大豆甙元对小鼠肝癌H22细胞的生长具有明显抑制作用,可诱导H22细胞凋亡,并在一定程度上抑制小鼠移植性肿瘤的生长,增强宿主免疫功能。
Objective To explore the effect of daidzein on the apoptosis of H22 tumor cells and its mechanism in mice. Methods The proliferation of H22 cells was detected by methyl thiazlyl tetrazolimm(MTT)assay.Sixty Kunming male mice were randomly divided into 6 groups(10 in each group).The mice in model and normal control group were given intraperitoneal injection of serum free RPMI 1640.The high,moderate,and low dose of daidzein groups were injected intraperitoneally with daidzein at different dose(50,100,and 200 mg/kg).The mice in positive control group were injected intraperitoneally with cyclophosphamide solution.After 14 days,the body weight,spleen index,thymus index,and tumor weight of the mice in different groups were compared.Bax and bcl-2 expressions in H22 cells were detected with western-blot. Results MTT assays showed that daidzein could inhibit the proliferation of H22 cells.After exposed to daidzein at a dose of 150 μmol/L for 72 hr,the proliferation inhibitory rate of the H22 cells was 30.3% (P<0.05).Compared to that of previous culture cycle,the growth inhibitory rate gradually increased.Thymus and spleen indexes in high and moderate dose daidzein groups were significantly higher(P<0.01).Compared with that of the model group,the tumor weight index of the three daidzein treatment groups decreased significantly(P<0.01 for all).Daidzein could regulated upward the expression of bcl-2 and downward the expression of bax.Compared with the 10.0μmol/L daidzein treatment group,the gray value for bax expression increased by 50.05% and that of bcl-2 expression decreased by 52.62%. Conclusion Daidzein inhibits the proliferation of H22 cells and induces apoptosis of H22 tumor cells.Daidzein can inhibit the growth of implanted tumor and promote the immune function of the mice.