RNA干扰抑制PDGF-α受体对人视网膜色素上皮细胞增殖的影响

Effect of PDGF-α receptor RNA interference on the proliferation of human retinal pigment epithelial cells

目的:探讨针对血小板源性生长因子α受体(PDGFR-α)的RNA干扰(RNAi)对体外培养的人视网膜色素上皮(h RPE)细胞增生的影响。方法:PDGFR-αsh RNA转染h RPE细胞48h后,MTT检测h RPE细胞的增殖,并计算转染物对细胞的抑制率,RT-PCR和Western blot检测h RPE细胞中PDGFR-α基因m RNA和蛋白的表达,流式细胞仪分析h RPE细胞的细胞周期。结果:h RPE细胞增殖明显受到抑制且呈剂量依赖性(P<0.05);PDGFR-α基因m RNA和蛋白的表达明显下调(P<0.05);G1期细胞百分比明显升高。结论:PDGFR-αsh RNA可沉默h RPE细胞中PDGFR-α基因m RNA和蛋白的表达,抑制细胞增殖。

Objective To investigate the effect of RNAi targeting PDGFR-α on the proliferation of the hRPE ceils cultured in vitro. Methods 48 h after PDGFR-α shRNA were transfected into hRPE cells,the proliferation of hRPE cells was detected by MTT experiment, and the inhibitory rate of PDGFR-α shRNA on proliferation was also measured. The expression of mRNA and protein of PDGFR-α gene in the cells was assayed by reverse transcription-PCR and Western blot, and the flow cytometry method was applied to detect the change of cell cycle of hRPE cells. Results The proliferation of hRPE cells was significantly inhibited in a dose- dependent manner （P 〈 0.05）, and the expression of mRNA and protein of PDGFR-αgene was notably lowered （P 〈 0.05）. The ratio of G1 phase of cells significantly increased. Conclusion PDGFR-α shRNA can silence the expression of mRNA and protein of PDGFR-α gene in hRPE cells, and inhibit cell proliferation.