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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: Multidrug resistance of tumor cells is the main cause of chemotherapy failure, and its mechanism is the change of expression of multiple resistance-related genes. Objective: To explore the multidrug resistance mechanism and reversal strategy of digestive tract tumors. METHODS: HCPT-resistant colon cancer cell line SW1116/HCPT was established by drug concentration increasing induction method; cell growth curve was measured by cell counting method; cell cycle distribution was measured by flow cytometry; chromosome karyotype analysis was performed by cell chromosome staining. ; MTT colorimetric method for drug sensitivity test; polymerase chain reaction (PCR) - enzyme linked immunosorbent assay (ELISA) to detect cell telomerase activity; tumor drug resistance functional classification gene chip Screening differentially expressed resistance-associated genes. Results: Compared with the parental cells, there was no significant change in the growth curve of SW1116/HCPT cells; the resistance to HCPT was increased by 120.0-fold, and it was associated with doxorubicin (48.2-fold), oxymatrine (2.1-fold), and arabinose. Cross-resistance was observed in glycosides (2.0-fold), 5-fluorouracil (1.8-fold), and cisplatin (1.3-fold); cell cycle distribution changed significantly, G1 phase cells increased, S phase cells decreased, and cell chromosomes did not change significantly. Karyotype There was no significant change in telomerase activity in aneuploidy 54-69. Function-based gene microarray showed that there were 30 genes with altered expression in resistant cell lines, of which 10 were down-regulated and 20 were up-regulated. Conclusion: SW1116/HCPT is one of the models for studying multidrug resistance in digestive tract tumors.