RNA干扰血管内皮生长因子受体2基因表达提高结肠癌细胞对奥沙利铂敏感性的研究

Study on the sensitivity of colon cancer cells for Oxaliplatin enhanced by RNA interference in the expression of vascular endothelial growth factor receptor-2

目的研究RNA干扰血管内皮生长因子受体2(VEGFR-2)基因表达对人结肠癌细胞生长的抑制作用和对奥沙利铂的化疗增敏作用。方法构建靶向VEGFR-2基因的小分子干扰RNA(siRNA)表达质粒并转染人结肠癌细胞株SW480;通过RT-PCR和Western blot方法检测VEGFR-2 siRNA对人结肠癌细胞株SW480 VEGFR-2mRNA和蛋白表达的影响;采用MTT实验和流式细胞术检测该siRNA及协同奥沙利铂对细胞增殖及凋亡的影响。结果 VEGFR-2 siRNA显著下调结肠癌细胞VEGFR-2 mRNA和蛋白的表达(P<0.05)。VEGFR-2 siRNA协同奥沙利铂使结肠癌细胞的生长变得更加缓慢,增殖受到明显抑制,细胞抑制及凋亡明显增强(P<0.05)。结论RNA干扰VEGFR-2基因表达可以抑制人结肠癌细胞株SW480的增殖,促进其凋亡,并能提高结肠癌细胞对奥沙利铂的敏感性。

Objective To study the inhibitory effect of RNA interference in the expression of vascular endothelial growth factor receptor-2（VEGFR-2） for the growth of human colon cancer cells and the chemosensitive effect of Oxaliplatin.Methods The expression plasmid of small interferon RNA（si RNA） targeting VEGFR-2 gene was constructed and human colon cancer cell line SW480 was transfected. The effect of VEGFR-2 si RNA for the expression of human colon cancer cells SW480 VEGFR-2 m RNA and protein was detected by RT-PCR and Western blot. The effect of si RNA or combining with Oxaliplatin for the cell proliferation and apoptosis was detected by MTT assay and flow cytometry. Results The m RNA and protein expression of VEGFR-2 were significantly down-regulated by VEGFR-2 si RNA（P 0.05）. The combination of VEGFR-2 si RNA and Oxaliplatin could slowdown the growth of colon cancer cells, inhibit the proliferation, and significantly increase the inhibition of cell proliferation and apoptosis（P 0.05）. Conclusion Silencing VEGFR-2gene by the si RNA technology can inhibit the proliferation of SW480 cells, induce the apoptosis of SW480 cell, and enhance the chemotherapy sensitivity of Oxaliplatin for human colon cancer cells.