芪术抗癌方及其联合5-氟尿嘧啶对CT26.WT结肠癌原位移植瘤模型小鼠肿瘤组织中凋亡相关蛋白表达的影响

and Its Combination with 5-Fluorouracil on the Expression of Apoptosis-related Proteins in Tumor Tissues of CT26.WT Colon Cancer Orthotopic Transplantation Tumor Model Mice

目的探讨芪术抗癌方抗结肠癌的可能作用机制。方法将90只BALB/c雄性小鼠随机分为空白组,模型组,5-氟尿嘧啶(5-Fu)组,芪术抗癌方高、中、低剂量组,联合高、中、低剂量组,每组10只。除空白组外其余各组小鼠建立CT26.WT结肠癌原位移植瘤模型。术后24h空白组与模型组予生理盐水10ml/(kg·d)灌胃;5-Fu组予注射用5-Fu 25mg/(kg·d)腹腔注射给药,隔日1次;芪术抗癌方高、中、低剂量组分别灌胃芪术抗癌方药液24、12、6g/(kg·d),每日1次;联合高、中、低剂量组分别灌胃芪术抗癌方相应剂量和5-Fu腹腔注射给药,用药剂量和方法同上。各组均干预3周后处死小鼠取肿瘤组织,称重并计算抑瘤率;检测各组小鼠肿瘤组织中半胱天冬氨酸蛋白水解酶3(Caspase-3)、B淋巴细胞瘤2(Bcl-2)及B淋巴细胞瘤2相关X蛋白(Bax)蛋白及mRNA表达。结果与模型组比较,其余各组小鼠瘤体质量均降低(P<0.01);联合高剂量组抑瘤率最高,达82.700%。各给药组对肿瘤组织中Caspase-3、Bax蛋白及mRNA表达均不同程度上调,而Bcl-2蛋白及mRNA表达下调,并且芪术抗癌方及其联合5-Fu对Caspase-3、Bax、Bcl-2蛋白及mRNA表达调节呈一定剂量依赖性(P<0.05或P<0.01)。结论芪术抗癌方可以抑制CT26.WT原位移植瘤的增殖,且与5-Fu同用具有增效作用,其机制可能与上调肿瘤组织中Caspase-3、Bax表达及下调Bcl-2表达有关。

Objective To explore the effects and possible mechanisms of Qizhu Kangai Formula(芪术抗癌方)on colon carcinoma.Methods A total of 90 male BALB/c mice were randomly divided into a blank group,a model group,a 5-fluorouracil(5-Fu)group,Qizhu Kangai Formula high,medium and low dose group,combined high,medium and low dose group,with 10 mice in each group.Except for the blank group,CT26.WT colon cancer orthotopic transplantation models of mice were established.Twenty-four hours after operation,the blank group and the model group were given 10 ml/(kg·d)of normal saline for gavage;the 5-Fu group was given intraperitoneal injection of 5-fluorouracil 25 mg/(kg·d)once every other day;Qizhu Kangai Formula high-dose,medium-dose,and low-dose groups were fed with Qizhu Kangai Fang 24,12,6 g/(kg·d)liquid solution once a day;Combined high-dose,middle-dose,and low-dose groups,were administered the corresponding doses of Qizhu Kangai Fang and 5-fluorouracil intraperitoneally.The dosages and methods were the same as above.Three weeks after intervention,the mice in all groups were killed and tumor tissues were taken to weigh and calculate the tumor inhibition rate;Caspase-3 and B lymphoma 2(Bcl-2)and blymphoma2 related Bax protein(Bax)and mRNA expression in each group were detected.Results Compared with the model group,the tumor mass weight of the mice in other groups decreased(P<0.01);the combined high-dose group had the highest tumor inhibition rate,reaching 82.700%.All administration groups up-regulated the expression of Caspase-3,Bax protein and mRNA in tumor tissue to varying degree,while the expression of Bcl-2 protein and mRNA was down-regulated.The inhibition and regulation effect of Qizhu Kangai Formula and combination with 5-fluorouracil on Caspase-3,Bax,Bcl-2 protein and mRNA expression was dose-dependent(P<0.05 or P<0.01).Conclusion Qizhu Kangai Formula could inhibit the proliferation of CT26.WT orthotopic transplanted tumors,and it has a synergistic effects with 5-Fu.The mechanisms may be related to up-regulation of Caspase-3 and Bax expression and down-regulation of Bcl-2 expression in tumor tissues.