奥沙利铂体外干预结直肠癌细胞株HT-29的耐药基因及相关蛋白的表达

Expression of drug resistance genes and their proteins in colorectal cancer cell line HT-29 after oxaliplatin intervention

目的:检测结直肠癌(colorectal cancer,CRC)奥沙利铂(oxaliplatin,L-OHP)耐药细胞株HT-29/L-OHP中多药耐药基因(multidrug resistance gene 1,MDR-1)、多药耐药相关蛋白的基因及他们编码的蛋白P-糖蛋白(P-glycoprotein,P-gp)、多药耐药相关蛋白(multidrug resistance associated protein,MRP)的表达,探讨其在L-OHP耐药中的作用。方法:利用MTT法检测L-OHP对原代细胞HT-29和耐药细胞HT-29/L-OHP的半数抑制率(IC50);利用RT-PCR及蛋白质印迹法检测HT-29/L-OHP中耐药基因及蛋白的表达情况。结果:L-OHP对HT-29和HT-29/L-OHP细胞的IC50分别为(4.15±0.17)μg.ml-1和(32.01±1.87)μg.ml-1,耐药指数为7.71;与原代细胞相比,HT29/L-OHP中耐药基因及蛋白表达均有所增加。结论:结直肠癌耐药细胞株中MDR-1、MRP基因及其编码的蛋白P-gp、MRP的表达高于原代细胞,这为预测CRC的多药耐药提供了新的依据。

Objective： To detect the differential expression of multidrug resistance gene 1,multidrug resistance associated protein gene and their proteins P-gp,MRP in colorectal cancer（CRC）HT-29/L-OHP cell line and its parental cell,in order to investigate their fuctions in L-OHP drug-resistance.Methods： IC50 of L-OHP in HT-29 and HT-29/L-OHP cell lines were detected by MTT;RT-PCR was used to exam the expressions of MDR-1 and MRP gene;Western blotting was used to exam the expressions of P-gp and MRP.Results： IC50 of L-OHP in HT-29 was（4.15±0.17）μg·ml^-1,IC50 of L-OHP in HT-29/L-OHP was（32.01±1.87） μg·ml^-1,the resistance index was 7.71.The expression levels of multidrug resistance genes and proteins in HT-29/L-OHP were higher than in HT-29 cell line.Conclusion： The expression levels of MDR-1,MRP gene,P-gp and MRP in HT-29/L-OHP were higher than in HT-29 cell line,detecting their expressions may predict the multidrug resistance in CRC.