2006•Zhonghua miniao waike zazhiRequires access
The effects of protein kinase C alpha cDNA on the expression of genes of multidrug resistance in renal cell carcinoma 786-0 cell line
Lizhuo Li
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Abstract
Objective To explore the effects of protein kinase C alpha(PKCα) cDNA on the expression of genes of multidrug resistance (MDR) in renal cell carcinoma(RCC) 786-0 cell line.Methods LR/BR recombination reactions were used to generate mammalian expression vectors of PKCα cDNA with C-terminal fused green fluorescent protein(GFP),and vectors were transfected into human RCC cell with Lipofectimine 2000.Western blot method and inverted fluorescent microscope were used to determine the expression of PKCα in RCC cells transfected by PKCα cDNA.RT-PCR was used to determine the expression of MDR-related genes MDR1,MRP1 and LRP in RCC cells transfected by PKCα cDNA.Results After transfection of 786-0 cells with pcDNA-DEST47-PKCα-GFP vectors,the results of Western blot showed that PKCα was highly expressed in human RCC 786-0 cells;and inverted fluorescent microscopy showed that GFP was highly expressed in RCC 786-0 cells.The results of semi-quantitative RT-PCR analysis showed that the expression level of MDR1 was higher in RCC cells transfected by PKCα cDNA than in RCC cells.Conclusions The expression of MDR1 mRNA in 786-0 cell line can be up-regulated by PKCα cDNA,which suggests PKCα cDNA can induce the increase of MDR in renal cell carcinoma.