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目的 探讨多药耐药蛋白P 170拮抗剂维拉帕米对胰腺癌细胞株化疗耐药的逆转作用。方法 采用RT PCR法对胰腺癌细胞株SW 1990和CAPAN 1中多药耐药基因MDR1mRNA的表达进行半定量分析 ;罗丹明外排试验检测胰腺癌细胞株中P 170的功能 ;MTT比色法评估胰腺癌细胞株对化疗药物的敏感性 ,并比较单纯化疗与化疗联合维拉帕米对其的疗效差异。结果 RT PCR检测结果显示MDR1mRNA在SW 1990中的表达丰度为 0 5 75 0± 0 0 76 4 ,在CAPAN 1中的表达丰度为 0 186 4± 0 0 5 36 ,两者具有显著差异 (P <0 0 1)。罗丹明外排试验表明 :SW1990阳性细胞百分数明显低于CAPAN 1(P <0 0 1) ;维拉帕米可使SW 1990阳性细胞百分数明显增加 (P <0 0 1)。MTT结果表明 :SW1990较CAPAN 1对ADM、MMC耐药 ;维拉帕米可部分逆转其对ADM和MMC的耐药性。结论 MDR1可能参与胰腺癌细胞株化疗耐药 ;中等剂量的维拉帕米联合化疗可逆转具有MDR1表型的人胰腺癌细胞株对ADM和MMC的耐药现象。
Objective To investigate the reversal effect of verapamil, a multidrug-resistant protein P 170 antagonist, on the chemoresistance of pancreatic cancer cell lines. Methods The expression of multidrug resistance gene MDR1 mRNA in pancreatic cancer cell line SW 1990 and CAPAN 1 was semi-quantitatively analyzed by RT-PCR. Rhodamine efflux assay was used to detect the function of P 170 in pancreatic cancer cell lines. MTT colorimetric assay Pancreatic cancer cell lines chemosensitivity, and compare the chemotherapy and chemotherapy combined with verapamil efficacy differences. Results The RT-PCR results showed that the expression abundance of MDR1 mRNA in SW 1990 was 0 5 75 0 ± 0 0 76 4 and that in CAPAN 1 was 0 186 4 ± 0 0 5 36 P <0 0 1). Rhodamine efflux assay showed that the percentage of SW1990 positive cells was significantly lower than CAPAN 1 (P <0.01); verapamil significantly increased the percentage of SW 1990 positive cells (P <0.01). The results of MTT showed that SW1990 was more resistant to ADM and MMC than CAPAN 1, and verapamil partially reversed its resistance to ADM and MMC. Conclusion MDR1 may be involved in chemoresistance in pancreatic cancer cell lines. Median dose of verapamil combined with chemotherapy may reverse the drug resistance of ADM and MMC in human pancreatic cancer cell lines with MDR1 phenotype.