Effect of p38 siRNA on biological behavior and sensitivities to sunitinib of human renal carcinoma cell line 786-O / 第二军医大学学报

ABSTRACT

Objective To investigate the effects of p38 gene sliencing on the proliferation, invasion, cell cycle and sensitivity to sunitinib of human renal carcinoma cell line 786-O. Methods We designed two sequence-specific small interfering RNA(siRNA531 and siRNA659) targeting p38 gene and transfected them into renal carcinoma cell line 786-0. 786-0 cells transfected with nonsense siRNA served asnegative control and those cultured with transfection medium served as blank control. The change of #38 gene expression was observed by RT-PCR and the expression of p38 protein was detected by Western blotting analysis. The proliferation, sensitivities to sunitinib, invasion capabilities, and the cell cycle of 786-0 cells were examined by CCK-8 assay, transwell chamber test and flow cytometry, respectively. Results RT-PCR and Western blotting analysis revealed that p38 expression in #38 siRNA group was significantly decreased compared with the controls. The cell proliferation rates were also significantly decreased 3-5 days after siRNA531 or siRNA659 transfection compared with the controls (P<0. 05, P<0. 01), and cells in the siRNA531 and siRNA659 groups become more sensitive to sunitinib compared with negative control group, with two IC50 values being significantly lower than that of the negative control group ([3. 2 ± 0. 3,, [1. 4±0. 1, μmol/mL vs [5. 4 ± 0. 2, μmol/mL; P<0. 05). In addition, analysis of cell cycle demonstrated a marked G50/G50 arrest of the 786-0 cells transfected with siRNA531 or siRNA659. We also noticed that 24 h after transfection, the cell invasion capabilities was significantly decreased in siRNA531 and siRNA659 compared with negative control (numbers of cells permeating septum 56. 43 ± 6. 02, 34. 00 ± 8. 12 vs 76. 27 ± 5. 08; P<0. 01). Conclusion We have successfully suppressed #38 gene expression by specific siRNA, which can inhibit the proliferation and invasion of human renal carcinoma cell line 786-0 and increase its sensitivity to sunitinib, paving a way for future treatment and targeted drug resistance of renal cell carcinoma.