摘要
目的探讨重组东亚钳蝎镇痛抗肿瘤肽(rAGAP)对胆管癌细胞化疗的增敏作用及其机制。方法体外培养人胆管癌细胞株HuCC-T1、QBC939和胆管上皮细胞HIBepic,单独用药:不同浓度的rAGAP、顺铂(DDP)和5-氟尿嘧啶(5-FU)处理三株细胞,四甲基偶氮唑盐比色法(MTT法)观察细胞的增殖抑制率及半抑制浓度(IC50)。联合用药:选用rAGAP(5μg/ml)、DDP(4μg/ml)、5-FU(50μg/ml)处理三株细胞,分为对照(A)组、rAGAP(B)组、DDP(C)组、rAGAP与DDP联用(D)组、5-FU(E)组、rAGAP与5-FU联用(F)组。MTT法检测细胞存活率;荧光定量PCR方法检测细胞多药耐药基因1(MDR-1)及多药耐药相关蛋白1(MRP-1)的mRNA表达。结果单独用药结果显示rAGAP抑制细胞增殖呈时间剂量依赖性,且对癌细胞具有一定的选择性;rAGAP组的IC50值明显小于DDP组和5-FU组(P<0.01)。联合用药结果显示,胆管癌细胞株HuCC-T1和QBC939中,D组较B、C组以及F组较B、E组细胞存活减少;而且MDR-1和MRP-1表达下调(P<0.01)。而在胆管上皮细胞HIBepic细胞系里,联合用药并未表现明显的增加化疗药效果的作用。结论 rAGAP可选择性抑制胆管癌细胞,与化疗药DDP和5-FU联用可增强化疗敏感性。
Objective To investigate the sensitivity-increasing effect of recommbinant analgesicantitumor peptide （rAGAP）on cholangiocarcinoma ceils to chemotherapy and its underlying mechanism. Methods The cholangioearcinoma cell lines HuCC-T1, QBC939 and bile duct epithelial cells HIBepic were cultured in vitro, and then exposed to rAGAP, eisplatin and 5-FU, respectively. The cells inhibition rate and 50 % inhibiting concentration（ICs0 ） were detected. The cells were divided into 6 groups of A（control）, B（treated with rAGAP 5μg/ml）, C（treated with DDP 4 μg/ml）, D（treated with 5-FU 50μg/ml plus DDP）, E（treated with 5-FU 50μg/ml） and F（treated with rAGAP plus 5- FU）. The cell survival rates were detected using MTT method and the mRNA expressions of multi- drug resistance-1 （MDR-1） and multidrug resistance protein-1 （MRP-1） were detected using RT-PCR Results Cholangiocarcinoma cells were time- and dosedependently inhibited by rAGAP which was selective for cancer. The ICs0 of rAGAP group was less than that of DDP and 5-FU groups （P〈0.01）. In H uCC-T1 and QBC939 cell lines, the cell survival rate was lower in group D than that in groups of B and C, which was lower in group F than that in groups of B and E（P〈0. 01）. Furthermore, the expressions of MDR-1 and MRP-1 were down-regulated（P〈0. 01）. The sensitivityincreasing effect of rAGAP on HIBepic cells to chemotherapy with combined use of drugs was not better than that with single drug. Conclusion rAGAP can inhibit cholangiocarcinoma cells growth and enhance the chemotherapy sensitivity when combined with cisplatin and 5-FU.
出处
《江苏医药》
CAS
北大核心
2013年第11期1241-1244,共4页
Jiangsu Medical Journal
基金
江苏省科技厅自然科学课题(BK2012871)
