人结肠癌羟基喜树碱耐药细胞株SW1116/HCPT的建立与鉴定

Construction and Characterization of Hydroxycamptothecin-resistant Human Colon Cancer Cell Line SW1116/HCPT

背景:肿瘤细胞的多药耐药性是造成化疗失败的主要原因,其机制是多种耐药相关基因表达的改变。目的:探讨消化道肿瘤的多药耐药机制和逆转策略。方法:应用药物浓度递增诱导法建立羟基喜树碱(HCPT)耐药结肠癌细胞株SW1116/HCPT;细胞计数法测定细胞生长曲线;流式细胞仪测定细胞周期分布;细胞染色体染色进行核型分析;四甲基偶氮唑蓝(MTT)比色法进行药物敏感实验;聚合酶链反应(PCR)-酶联免疫吸附测定(ELISA)检测细胞端粒酶活性;肿瘤药物耐受功能分类基因芯片筛选差异表达的耐药相关基因。结果:与亲代细胞相比,SW1116/HCPT细胞生长曲线无明显改变;对HCPT的耐药倍数增加120.0倍,并与阿霉素(48.2倍)、氧化苦参碱(2.1倍)、阿糖胞苷(2.0倍)、5-氟尿嘧啶(1.8倍)、顺铂(1.3倍)存在交叉耐药现象;细胞周期分布明显改变,G1期细胞增加,S期细胞减少;细胞染色体无明显变化,核型为非整倍体54-69;细胞端粒酶活性无明显改变;功能分类基因芯片显示,耐药细胞株表达改变的基因有30个,其中表达下调10个,上调20个。结论:SW1116/HCPT是研究消化道肿瘤多药耐药现象的模型之一。

Background： One of the main reasons for the failure of chemotherapy is multi-drug resistance of cancer cells. There are many kinds of drug resistant genes whose expression levels have altered. Aims： To study the multi-drug resistance mechanism of digestive tract tumors and its reverse strategy. Methods： Stepwise concentration-increasing method was used to construct SWI 116/HCPT which was a hydroxycamptothecin-resistant cell line of human colon cancer. Cell counting, flow cytometry, chromosome staining, methyl thiazolyl tetrazolium （MTT） analysis and polymerase chain reaction （PCR）-enzyme-hnked immunosorbent assay （ELISA） protocol were used to determine the cell growth curve, cell cycle distribution, karyotype analysis, drug sensitivity test and telomerase activity, respectively. Human cancer drug resistance ＆ metabolism gene array were used to determine the expression profile of drug resistant genes. Results： Compared with their parent cells, the cell growth curve of SW1116/HCPT had no obvious change. SW1116/HCPT was 120.0 times more resistant to cytotoxic action of hydroxycamptothecin and had cross-resistance to adriamycin （48.2）, oxymatrine （2.1）, arabinosylcytosin （2.0）, 5-FU （1.8） and CDDP （1.3）. The number of SW116/HCPT cells in GI phase increased and that of S phase decreased and was statistically significant （P〈0.01）. Karyotype and telomerase activity of SW116/HCPT cells had no apparent changes comparing with their parent cells. The karyotype was aneuploid 54-69. There were 30 drug resistant genes whose expression levels were altered. 10 were down-regulated and 20 up-regulated. Conclusions： SWI 116/HCPT can be one of the models for the study of multi-drug resistance in digestive tract tumors.