中文摘要：

目的：建立人结肠癌奥沙利铂（oxaliplatin,L-OHP）耐药细胞HCT116/L-OHP,并初步探讨其可能的耐药机制。方法：通过逐步增加作用于亲代细胞HCT116的L-OHP浓度与间断大剂量L-OHP作用,建立耐药细胞HCT116/L-OHP;MTT法检测L-OHP、5-氟尿嘧啶（5-fluorouracil,5-FU）、顺铂（cisplatin）和7-乙基-10-羟基喜树碱（7-ethyl-10-hydrol-camptothecin,HCPT）对HCT116和HCT116/L-OHP细胞的细胞毒性;蛋白质印迹法检测相关耐药蛋白的表达;基因芯片检测信号通路的改变。结果：成功构建了稳定耐药的耐药细胞HCT116/L-OHP,耐药倍数为17倍,与5-FU、DDP和HCPT有不同程度的交叉耐药性。与HCT116细胞比较,HCT116/L-OHP细胞中P糖蛋白（P-glycoprotein,P-gp）、肺耐药蛋白（lung resistance protein,LRP）和谷胱甘肽S转移酶（glutathione S-transferase,GST）的表达上调（P〈0.01）,9条信号通路上调（P〈0.05）,其中细胞周期信号通路上调最明显,p53信号通路次之。结论：HCT116/L-OHP细胞具有稳定的耐药性,其耐药机制可能与细胞耐药蛋白表达、细胞周期和p53信号通路表达上调有关。

英文摘要：

Objective： To establish an oxaliplatin （L-OHP）-resistant human colon carcinoma HCT116/L-OHP cell, and to preliminarily explore the mechanism of resistance. Methods： Oxaliplatin-resistant HCT116/L-OHP cells were established by gradually increasing the concentration of L-OHP and intermittent treatment with high-dose concentration on parental cells （HCT116）. The cytotoxicities in HCT116 and HCT116/L-OHP cells induced by L-OHP, 5-fluorouracil （5-FU）, cisplatin （DDP） and 7-ethyl-10-hydrol-camptothecin （HCPT） were detected by MTT assay. The expressions of resistance-associated proteins were determined by Western blotting. The changes of signaling pathway were measured by gene chips. Results： The HCT116/L-OHP cells with stable resistance were successfully established. The resistance fold was 17. The HCT116/L-OHP cells had cross-resistance with 5-FU, DDP and HCPT. The expression levels of permeability-glycoprotein （P-gp）, lung resistance protein （LRP） and glutathione S-transferase （GST） were higher in HCT116/L-OHP cells than those in HCT116 cells （P0.01）. There were nine signaling pathways up-regulated （P0.05）. The most obviously up-regulated signaling pathway was cell cycle signaling pathway, and the next was the p53 signaling pathway. Conclusion： The mechanism of HCT116/L-OHP cells with stable resistance may be related with the up-regulation of resistance-associated protein expression, the cell cycle and the p53 signaling pathway.