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目的探讨蛋白激酶 C 抑制剂对肾癌细胞多药耐药性的逆转作用的机制。方法应用荧光显色法、RT- PCR、Western Blot 方法检测 PKCαcDNA 对肾癌 786- 0 细胞的转染前后细胞中 MDR 相关基因MDR1、MRP1、LRP 的表达变化。采用 MTT 法测定转染细胞系 PKC- α786- 0 与 786- 0 细胞分别对阿霉素( ADM) 与蛋白激酶 C 激动剂、蛋白激酶 C 抑制剂协同作用后的耐药性变化。结果 RT- PCR 结果显示肾癌转染细胞 PKCα786- 0 的 MDR1 表达水平高于肾癌 786- 0 细胞。阿霉素( ADM) 与蛋白激酶 C 抑制剂协同作用的细胞系的耐药性明显降低。786- 0 细胞对阿霉素( ADM) 的 IC50 为: 7.8015e-7(5.7046e-7 至 1.0669e-6);PKCα786- 0 对药物阿霉素( ADM) 的 IC50 为: 1.6588e-6(1.1621e-6 至 2.3677e-6); 蛋白激酶 C 激动剂 PMA 联合阿霉素处理 PKCα786- 0 的 IC50 为: 2.6794e-6(2.0521e-6 至 3.4983e-6); 蛋白激酶 C 抑制剂 Calphostin C 联合阿霉素处理 PKCα786- 0 的 IC50 为: 9.2506e-8(5.9337e-8～1.4422e-7)。结论蛋白激酶 C 抑制剂可以逆转人肾癌细胞的多药耐药性, 其途经可能与改变 MDR1 的表达相关。
Objective To investigate the mechanism of reversal of multidrug resistance in renal cell carcinoma by protein kinase C inhibitor. Methods The expression of MDR-related genes MDR1, MRP1, LRP in human renal carcinoma 786- 0 cells before and after transfection were detected by fluorescent chromometry, RT-PCR and Western Blot. MTT assay was used to determine the drug resistance changes of transfected cell lines PKC-α | 786-0 and 786-0 cells to doxorubicin (ADM) in combination with protein kinase C agonist and protein kinase C inhibitor respectively. Results The results of RT-PCR showed that the expression level of MDR1 in PKCα 786- 0 cells was higher than that in renal carcinoma 786-0 cells. The drug resistance of the cell lines that act in synergy with doxorubicin (ADM) and protein kinase C inhibitors is significantly reduced. The IC50 for 786-0 cells for doxorubicin (ADM) was 7.8015e-7 (5.7046e-7 to 1.0669e-6); the IC50 for PKCα 786-0 for drug doxorubicin (ADM) was 1.6588 (1.1621e-6 to 2.3677e-6); IC50 for protein kinase C agonist PMA in combination with doxorubicin treatment PKCα 786-0 was 2.6794e-6 (2.0521e-6 to 3.4983e-6 ); IC50 of protein kinase C inhibitor Calphostin C combined with doxorubicin treatment PKCα 786- 0 was: 9.2506e-8 (5.9337e-8 ~ 1.4422e-7). Conclusions Protein kinase C inhibitor can reverse the multidrug resistance of human renal cell carcinoma cells, and its possible mechanism is related to the change of MDR1 expression.