摘要
目的探讨蛋白激酶C（PKC）抑制剂对肾癌细胞多药耐药（MDR）逆转作用的机制。方法应用荧光显色法、RT-PCR和Western blot方法，检测PKCα cDNA转染肾癌786-0细胞前后，细胞中多药耐药基因1（MDR1）、多药耐药相关蛋白1（MRP1）和肺耐药蛋白（LRP）表达的变化。采用MTT方法，分别测定阿霉素（ADM）与PKC激动剂以及与PKC抑制剂协同作用后，786-0细胞和肾癌转染细胞系PKCα／786-0耐药性的变化。结果RT—PCR结果显示，肾癌转染细胞系PKCα／786-0的MDR1表达水平高于肾癌786-0细胞。ADM与PKC抑制剂协同作用的PKCα／786-0细胞系耐药性明显降低。786-0细胞对ADM的IC50为7．8015e^-7（5．7046e^-7～1．0669e^-6）；PKCα／786-0对ADM的IC50为1．6588e^-6（1．1621e^-6～2．3677e^-6）。PKC激动剂佛波醇酯（PMA）联合ADM处理PKCα／786-0的IC50为2．6794e^-6（2．0521e^-6-3．4983e^-6）；PKC抑制剂Calphostin C联合ADM处理PKCα／786-0的IC50为9．2506e^-8（5．9337e^-8～1．4422e^-7）。结论PKC抑制剂可以逆转入肾癌细胞的多药耐药性，其途径可能与改变MDR1的表达有关。
Objective To explore the mechanism of reversal of multidrug resistance in renal carcinoma cells by protein kinase C inhibitor. Methods RT-PCR, Western blot and inverted fluorescent microscopy were used to determine the expression of PKCα and MDR related gene MDR1, MRP1, LRP in RCC cells transferred by PKCα cDNA. Also effects of activator and inhibitor of PKC in combination with adriamycin on muhidrug resistance in RCC cells were evaluated by MTT. Results The results of semiquantitative RT-PCR analysis showed that the expression level of MDR1 was higher in RCC cells transferred by PKCα cDNA than in RCC cells, the reversal effectiveness of PKC inhibitors in combination with adriamycin （ADM）was apparently favorable. IC50 of ADM in 786-0 cells was 7. 8015e^-7 （5. 7046e^-7 to 1. 0669e^-6 ） ; IC50 of ADM in PKCα/786-0 cells was 1. 6588e^-6 （ 1. 1621e^-6 to 2. 3677e^-6 ） ; IC50 of ADM in combination with PMA in PKCα/786-0 cells was 2.6794e^-6 （2.0521e^-6 to 3. 4983e^-6 ） ; IC50 of ADM in combination with calphostin C in PKCα/786-0 cells was 9.2506e^-8 （5. 9337e^-8 to 1. 4422e^-7 ）. Conclusion PKC inhibitors can reverse multidrug resistance in renal carcinoma cells in vitro via changes of expression of MDR1.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2006年第2期92-95,共4页
Chinese Journal of Oncology
基金
辽宁省科学技术基金资助项目(9910500107)
关键词
蛋白激酶C
肾癌细胞
多药耐药
Protein kinase C
Renal cell carcinoma
Multidrug resistance
