三氧化二砷联合5-氟尿嘧啶对小鼠肝癌H_(22)细胞移植瘤的作用

Research of the effects of arsenic trioxide combined with 5-fluorouracil on mice liver cancer cell line H_(22) subcutaneously implanted liver cancer in vivo

目的探讨三氧化二砷(As2O3)联合5-氟尿嘧啶(5-FU)对小鼠肝癌H22细胞皮下移植瘤的作用,为提高治疗肝癌的临床效果、降低毒性和克服耐药提供理论和实验依据。方法建立昆明小鼠肝癌皮下移植瘤动物模型,按瘤体体积均衡分为4组进行干预,依次为对照组(NS)、As2O3组(0.2mg/kg),5-FU组(10 mg/kg),As2O3(0.2 mg/kg)+5-FU(10 mg/kg)组,每组10只,连续腹腔注射药物2周,停药后第2天处死动物,留取血液标本,解剖提取肿瘤标本。分别测定比较各组小鼠肿瘤体积、肿瘤组织病理变化、肿瘤细胞增殖情况、凋亡率及肝肾功能相关生化指标。结果各组肿瘤体积有明显差异(F=252.7,P<0.01),各药物干预组平均肿瘤体积均明显低于对照组(P<0.01),同时As2O3+5-FU组平均肿瘤体积显著低于单用As2O3和5-FU组(P<0.01)。As2O3组和5-FU组细胞增殖指数显著低于对照组(P<0.05),As2O3+5-FU组细胞增殖指数显著低于对照组和As2O3组(P<0.05),但与5-FU组比较无显著差异(P>0.05);As2O3组细胞凋亡率高于对照组(P<0.05),As2O3+5-FU组细胞凋亡率显著高于As2O3组和5-FU组(P<0.05),血清ALT、AST、Cr检测值各组间无显著性差异(P>0.05)。结论体内As2O3和5-FU联合应用能显著增强对小鼠肝癌皮下移植瘤的抑制作用,两者具有协同作用,机制可能与增强抑制细胞增殖和诱导凋亡有关。且两者联合使用并未发现明显毒副作用。

Objective To evaluate the effects of arsenic trioxide（As2O3） combined with 5-fluorouracil（5-FU） on mouse hepatocarcinoma H22 subcutaneously transplanted tumor and provide the theoretical and experimental evidence for improving the clinical effects,reducing toxicity and overcoming drug resistance in the treatment of hepatocellular carcinoma.Methods Treat hepatocarcinoma subcutaneously transplanted tumor in mice with As2O3 and 5-FU in vivo： hepatocarcinoma H22 subcutaneously transplanted tumor models were established in 24 mice which were divided into four groups based on the tumor volume with drug intervention： NS group,As2O3（0.2 mg/kg） group,5-FU（10 mg/kg） group and As2O3（0.2 mg/kg）＋5-FU（10 mg/kg） group.The mice was executed in the next day after discontinuing 2 weeks continuous intraperitoneal drug injection and blood and tumor specimens were conserved.The tumor volume was separately measured and compared,the tumor pathomorphological change after hematoxylin-eosin staining and the proliferating cell nuclear antigen（PCNA） were observed in each group.The transplantation of end labeling（TUNEL method） was used to observe the tumor cell proliferation situation,calculate apoptosis rate and test hepatic and renal functions relevant biochemical indexes in mice.Results The tumor volumes in each group had significant difference（F=252.7,P 0.01）.The average tumor volumes of the groups with drug intervention were significantly lower than that in NS group（P 0.01）.At the same time,the average tumor volume of As2O3＋5-FU group was significantly lower than those of groups with As2O3 or 5-FU alone（P0.01）.The cellproliferation index in As2O3 group was significantly lower than that in NS group（P0.05）.The cell prolif-eration index in As2O3＋5-FU group was significantly lower than those in NS group and As2O3group（P0.05）,less than those in 5-FU group but no significant difference with it（P0.05）.The apoptosis rate of the tumor in As2O3group was higher than that in NS group with statistical significance（P0.05）.The apoptosis rate of thetumor in As2O3＋5-FU group was significantly higher than those in As2O3 group and 5-FU group withstatistical significance（P0.05）.The measurement values of serum ALT,AST,Cr in each groups had nosignificant difference（P0.05）.Conclusion The combined application of As2O3 and 5-FU can significantlyenhance the inhibiting effect of mouse hepatocarcinoma subcutaneously transplanted tumor.Both of them had syner-gistic effect with the mechanism related to the enhancement on the inhibition of cell proliferation and induction of apoptosis.Also little side effects are found.