地锦草黄酮醇诱导U14宫颈癌小鼠肿瘤细胞凋亡作用及其机制

Effect and mechanism of flavonol from Euphorbia Humifusa Willd on the apoptosis of tumor cells in U14 cervical cancer mice

目的研究地锦草黄酮醇(Euphorbia Humifusa Willd Flavonol,EHWF)诱导U14宫颈癌小鼠肿瘤组织细胞凋亡作用和分子机制。方法将40只U14宫颈癌小鼠随机分成阴性对照组、环磷酰胺(CTX)阳性对照组、EHWF低剂量组和EHWF高剂量组四组,每组各10只。接种肿瘤24小时后,小鼠灌胃给药,EHWF剂量分别为100 mg/kg和200 mg/kg,连续给药14天。阴性对照组灌服蒸馏水,阳性对照组腹腔注射环磷酰胺(25 mg/kg)。第15天,取肿瘤组织。TUNEL染色检测细胞的凋亡情况;流式细胞仪分析小鼠肿瘤细胞周期与细胞凋亡关系;Western-blot分析Caspase-8、Caspase-3、CyclinD1和P16在凋亡过程中蛋白表达变化。结果TUNEL染色分析发现高剂量组肿瘤组织中凋亡细胞明显高于对照组。与对照组比较,细胞周期中的S期及G2/M期比例明显降低。凋亡相关蛋白Caspase-8、Caspase-3和P16在高剂量组中表达明显增加,而周期蛋白CyclinD1表达下调。结论EHWF参与诱导U14宫颈癌小鼠肿瘤细胞凋亡过程。

Objective To study the effect of Euphorbia Humifusa Willd Flavonol(EHWF) on apoptosis of tumor cells in U14 cervical cancer mice and its mechanism.Methods 40 U14 cervical cancer mice were randomly divided into 4 groups containing the negative control group,cyclophosphamide(CTX) group,low dose EHWF group and high dose EHWF group with 10 mice in each group.24 hours after inoculated with tumor cells,mice were orally administrate with 100 mg/kg and 200 mg/kg of EHWF.Mice in negative control group were intragastric administrated with distilled water and the positive control group were intraperitoneal injected with 25 mg/kg of CTX.The treatment were lasted for 14 days continuously.On day 15,tumor tissue was isolated frommice.TUNEL staining was used to detect cell apoptosis.The relationship between tumor cell cycle and apoptosiswas analyzed by flow cytometry.The protein expressions of Caspase-8,Caspase-3,CyclinD1 and P16 duringapoptosis were analyzed by Western-blot analysis.Results TUNEL staining analysis showed that the apoptoticcells in high-dose group were significantly higher than those in control group.Compared with the control group,theratio of S phase and G2/M phase in high dose group were significantly decreased.The expression of Caspase-8,Caspase-3 and P16 were increased in high dose group,and the expression level of Cyclin D1 was decreased(P<0.05).Conclusions EHWF is involved in inducing apoptosis of U14 cervical cancer mice.