TNP-470联合丝裂霉素对小鼠原位移植性肝癌生长抑制及作用机制的实验研究

Study of TNP-470 in the enhancement of anti-tumor effect of mitomycin on hepatocellular carcinoma

目的 研究血管形成抑制剂TNP 4 70联合MMC对小鼠原位移植性肝癌生长抑制及作用机制。方法 小鼠H2 2肝癌细胞先在KM小鼠皮下造成实体瘤 ,再原位接种于KM小鼠肝脏。原位肝癌模型小鼠 4 0只 ,分为四组。 (1)对照组 :隔日皮下注射溶剂 (3%酒精 ) ;(2 )MMC组 :一周 2次腹腔注射MMC(2mg/kg) ;(3)TNP 4 70组 :隔日皮下注射TNP 4 70 (30mg/kg) ;(4)TNP 4 70 +MMC组 :隔日皮下注射TNP 4 70 (30mg/kg) ,一周 2次腹腔注射MMC(2mg/kg)。用药 2周后处死小鼠 ,检测原位肿瘤的体积 ,治疗前后小鼠体重 ;肿瘤组织送检ki 6 7、凋亡及微血管密度 ;另取肿瘤组织电镜观察。结果 (1)各治疗组肿瘤生长均较对照组有不同程度的抑制 ,联合用药组最显著 (P <0 0 5 ) ;各组治疗前后体重变化无明显差异 ;(2 )相比于对照组 ,TNP 4 70对增殖指数无明显影响 (P >0 0 5 ) ,凋亡指数增加 ,微血管密度明显减少 (P <0 0 5 )。联合用药组增殖指数及凋亡指数较对照组及单用药组明显增加 ,微血管密度较对照组及MMC组明显减少 (P <0 0 5 )。 (3)电镜显示 :联合用药组可见凋亡的内皮细胞及较多凋亡的肿瘤细胞。结论 联合TNP 4 70及MMC能明显提高单药对小鼠原位移植瘤生长抑制 ,TNP 4 70作用的靶点主要是血管内皮细胞 。

Objective To investigate the effect of TNP-470 enhancing the anti-tumor function of mitomycin (MMC)and the mechanism.Methods Forty KM mice bearing hepatocellular carcinoma with orthotopic transplantation were divided randomly into 4 groups:①control(3% ethanol,sc,eod);②TNP-470(30mg/kg,sc,eod);③MMC(2mg/kg,ip,biw);④TNP-470(30mg/kg,sc,eod)+MMC(2mg/kg,ip,biw);Total mice were sacrificed two weeks after administration.Then the volume of the tumor,the pre-therapeutic and the post-therapeutic weights of the mice were studied and immunohistochemistry was employed to invesitgate ki-67,apoptosis,MVD;and also apoptosis was observed by the electron microscope.Results 1.the tumor growth of every therapeutic groups were significantly different from the control,and the most significant difference existed between the combination group and the control.( P< 0.01).The weight changes of the mice between every pre-therapeutic group and the post-therapeutic group did not have significant difference( P> 0.05);2.there was no significant difference in the PI between the TNP-470 group and the control( P> 0.05),but,on the contrary,in the AI and MVD( P< 0.01),the AI and PI of the combination group were significantly different from the other 3 groups( P< 0.01),the MVD of the combination group was decreased more than those of the control and MMC groups( P< 0.01);3.The result of electron microscope indicated that the more apoptotic endothelial cells and apoptotic tumor cells can be seen in the combination group than those in other groups.Conclusion TNP-470 can enhance the anti-tumor effect of mitomycin(MMC).The direct target of TNP-470 is the endothelial cell,and the mechanism of additive effect of TNP-470 may be through the way of inducing the apoptosis of the endothelial cell.