肠胃清对结肠癌肝转移裸鼠生存期及肿瘤组织铜转运相关蛋白表达的影响

Effect of Changweiqing on Survival Duration and Copper-transporting Tumor Tissues-related Protein Expression of Nude Mice in Colon Carcinoma Hepatic Metastasis Model

目的:观察肠胃清对结肠癌肝转移裸小鼠生存期、肿瘤生长及对肿瘤组织铜转运相关蛋白表达的影响,探讨肠胃清对结直肠癌化疗增效的作用机制。方法:BALB/C裸小鼠,采用HCT116细胞建立裸小鼠结肠癌肝转移模型,随机分为模型组,肠胃清组、化疗组、肠胃清联合化疗组(联合组),每组20只,肠胃清组ig给予相应药物,ip生理盐水,化疗组ip奥沙利铂(L-OHP),5-氟尿嘧啶(5-FU),ig给予生理盐水,联合组ig给予肠胃清,ip L-OHP,5-FU,模型组ig和ip生理盐水,连续给药3周。观察肠胃清对荷瘤小鼠生存期及肿瘤生长的影响;利用免疫组化观察肿瘤组织人铜离子转运蛋白1(h CTR1),铜转运蛋白α链(ATP7A)和P型铜转运ATP酶(ATP7B),多药耐药相关蛋白2(MRP2),谷胱甘肽S-转移酶-π(GST-π)蛋白表达的影响;采用TUNEL法观察移植瘤组织细胞凋亡率。结果:与模型组比较,肠胃清组、化疗组、联合组抑瘤率分别为45.33%,70.51%,72.07%,以联合组明显(P<0.01);肠胃清组、化疗组、联合组裸小鼠生存时间均长于模型组(P<0.05);肠胃清组、联合组肿瘤组织中h CTR1的表达阳性率均高于模型组和化疗组(P<0.05);肠胃清组、联合组ATP7A,ATP7B和GST-π的表达阳性率均低于模型组和化疗组(P<0.05);肠胃清组、联合组的MRP2表达阳性率低于模型组(P<0.05);联合组凋亡指数高于化疗组和模型组(P<0.01)。结论:肠胃清能上调肝转移瘤组织h CTR1的表达,下调ATP7A和ATP7B蛋白表达,降低GST-π和MRP2的表达,并有增加肿瘤组织内铂药物含量的趋势,同时能诱导细胞凋亡,增加肿瘤细胞凋亡率,从而提高化疗敏感性。

Objective： To study the effect of Changweiqing on survival duration,tumor growth and copper-transporting tumor tissues-related protein expression of nude mice in colon carcinoma liver metastasis model,in order to discuss the synergistic mechanism of Changweiqing on the chemotherapy for colorectal cancers.Method： BALB / C nude mice were selected to establish the nude mice colorectal cancer hepatic metastasis model and randomly divided into the model group, the Changweiqing group, the chemotherapy group and the Changweiqing plus chemotherapy（ combination） group,with 20 mice in each group. The Changweiqing group was orally administered with corresponding drugs and intraperitoneally injected with normal saline. The chemotherapy group was intraperitoneally injected with oxaliplation（ L-OHP） and 5-fluorouracil（ 5-FU） and orally administered with normal saline. The combination group was orally administered with Changweiqing and intraperitoneally injected with L-OHP and 5-FU; The model group was orally administered and intraperitoneally injected with normal saline. All group were treated for three weeks. The effect of the Changweiqing on survival duration and tumor growth of colorectal cancer hepatic metastasis in nude mice was observed. Human copper transporter 1（ h CTR1）,ATPase Cu2 ＋transporting alpha polypeptide（ ATP7A）, copper transporting P-type adenosine triphosphotase（ ATP7B）,multidrug resistanceassociated protein 2（ MRP2） and glutathione-S-transferase-π（ GST-π） protein expressions in tumor tissue were detected by the immunohistochemical method. The apoptosis rate of transplanted tumor tissues was observed by TUNEL method. Result： The Changweiqing group,the chemotherapy group and the Combination group revealed the tumor inhibition rate of 45. 33%,70. 51%,72. 07%,particularly the Combination group（ P 〈 0. 01）. Those group showed a longer nude mice survival time compared with the control group（ P 〈0. 05）. The Changweiqing group and the Combination group showed higher h CTR1 but lower ATP7 A,ATP7B,MRP2,GST-π protein expressions in nude mouse tumors than the model group and the chemotherapy group（ P 〈0. 05）. The Combination group displayed a higher apoptosis index than the chemotherapy group and the model group（ P 〈 0. 01）. Conclusion： The Changweiqing group can up-regulate the expression of h CTR1 in liver metastases tumor tissues, down-regulate ATP7 A and ATP7 B protein expressions, reduce GST-π and MRP2 expressions,increase platinum content in tumor tissues,induce apoptosis and increase the apoptosis rate,so as to enhance the chemosensitivity.