人结肠癌LS174T/5-Fu多药耐药细胞系的建立

Establishment of Human Colon Cancer Multi-Drug Resistant Cell Line LS174T/5-Fu

目的建立人结肠癌多药耐药细胞系,并研究其生物学特性。方法以5-氟尿嘧啶(5-Fu)为诱导药物,LS174T细胞株为研究对象,利用浓度递增诱导法,建立LS174T/5-Fu多药耐药细胞系。MTT法测定药物敏感性,光镜、细胞计数法、流式细胞仪及RT-PCR等方法观察其生物学特征及多药耐药相关基因(MRP)mRNA表达。结果历时6个月成功建立人结肠癌多药耐药细胞系LS174T/5-Fu,对5-Fu耐药指数为40.24,且与羟基喜树碱、顺铂、依托泊苷和脱氧氟尿苷等多种化疗药物有不同程度的交叉耐药性;细胞形态发生改变;倍增时间较亲代细胞延长;细胞周期发现其G1和S期细胞增加,G2期细胞减少;MRP mRNA表达增高。结论LS174T/5-Fu细胞是一种可靠的人结肠癌多药耐药细胞系,该细胞系的建立为深入研究5-Fu诱导人结肠癌多药耐药的发生机制奠定了基础。

Objective To establish a human colon cancer multi-drug resistant cell line LS174T/5-fluorouracil（5- Fu） and to explore its biological characteristics. Methods A resistant human colon cancer line LS174T/5-Fu was established by stepwise increasing concentrations of fluorouracil selection from the parental cell line LS174T. Drug sensitivity was detected by MTT assay, and the changes of biological characteristics were determined by light microscopy, cell counting,flow cytometry and RT-PCR. Results LS174T/5-Fu cell line was developed after 6 months with stable resistance to 5-Fu and a resistance index of 40.24. The cells exhibited cross-resistance to cisplatin, etoposide, doxifluridine and hydeoxycamptothecin. LS174T/5-Fu cells grew more slowly than LS174T cells, which was longer than LS174T cell line. Cell cycle distribution of LS174T/5-Fu cells was changed compared with parental cells. The percentage of cells in G1 and S phase was increased,while in G2 phase was decreased. The level of MRP mRNA expression was increased according to the concentration of 5-Fu in resistant cell lines. Conclusion LS174T/5-Fu cells is a reliable multi-drug resistant cell subline of human colon cancer. The successful establishment of this cell subline has laid a solid foundation for further study of the multi-drug resistant mechanism of human colon cancer induced by 5- fluorouracil.