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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 multidrug resistance gene (MDR-1) in oxaliplatin (L-OHP) resistant cell line HT-29/L-OHP in colorectal cancer (CRC) The expression of multidrug resistance-associated proteins and their encoded proteins P-glycoprotein (P-glycoprotein, P-gp) and multidrug resistance associated protein (MRP) are discussed in L- The role of OHP resistance. METHODS: The half inhibition rate (IC50) of L-OHP on primary cell HT-29 and drug resistant cell HT-29/L-OHP was detected by MTT assay; HT-29/L- was detected by RT-PCR and Western blotting. Resistance gene and protein expression in OHP. Results: The IC50 values of L-OHP to HT-29 and HT-29/L-OHP cells were (4.15 ± 0.17) μg.ml-1 and (32.01 ± 1.87) μg.ml-1, respectively, and the resistance index was 7.71; Compared with primary cells, the expression of drug resistance genes and proteins in HT29/L-OHP increased. Conclusion: The expression of MDR-1 and MRP genes and their encoding proteins P-gp and MRP are higher in colorectal cancer-resistant cell lines than in primary cells, which provides a new basis for predicting multidrug resistance of CRC.