靶向PDGF受体-α的RNA干扰对RPE细胞增殖和凋亡的影响  

Effects of RNA interference targeting PDGF receptor-α on RPE cells proliferation and apoptosis

作　　者：秦贤杰[1] 彭燕一[1] 秦程[1] 

机构地区：[1]桂林医学院附属医院眼科,桂林541000

出　　处：《安徽医科大学学报》2016年第2期180-184,共5页Acta Universitatis Medicinalis Anhui

基　　金：广西自然科学基金项目(编号:KY2012021);广西科学实验中心开放课题项目(编号:KFJJ2011-11)

摘　　要：目的探讨针对血小板源性生长因子受体-α（PDGFR-α）的RNA干扰对体外培养的人视网膜色素上皮（hRPE）细胞增殖和凋亡的影响，为临床治疗增殖性玻璃体视网膜病变（PVR）提供实验依据。方法利用脂质体将合成的shRNA转染体外培养的hRPE细胞，MTT法检测hRPE细胞增殖活性；RT-PCR法检测细胞中PDGFR—α mRNA的表达；Western blot法检测PDGFR-α蛋白表达水平；Hoeclast 33258荧光染料核染分析细胞凋亡；流式细胞仪检测细胞周期及凋亡率。结果PDGFR-α shRNA作用hRPE细胞24～72h后，细胞增殖明显受到抑制，呈时间剂量效应（P〈0．05）；PDGFR-α shRNA作用hRPE细胞48h后，实验组PDGFR—α基因mRNA和蛋白的表达水平较空白对照组及阴性对照组明显降低（P〈0．05）；实验组细胞的凋亡率分别为（37．10±0．55）％、（50．8±1．41）％，与空白对照组（11．7±1．11）％及阴性对照组（13．5±1．05）％相比，差异有统计学意义（P〈0．05）；实验组G1期细胞百分比分别为（64．76±1．16）％、（75．64±0．93）％，与空白对照组（54．51±1．40）％及阴性对照组（56．04±2．19）％相比，差异有统计学意义（P〈0．05）。结论PDGFR-α shRNA能抑制hRPE细胞的增殖并诱导其凋亡。Objective To investigate the effect of RNA interference targeting platelet-derived growth factor receptor-α （PDGFR-α） on hRPE cell proliferation and apoptosis in vitro, so that experimental evidence was provided for the clinical treatment of proliferative vitreoretinopathy. Methods Retinal pigment epithelium cells were cultured in vitro, and shRNA which was chemically synthesized was transfected into hRPE cell mediated by cationic liposomes, the cell proliferation ability was detected by MTT; the expression of PDGFR-α mRNA of hRPE cells was detected by Reverse Transcription Polymerase Chain Response; the expression of PDGFR-α protein was detected by Western blot ; cell apoptosis was analyzed by Hochest 33258 ; the Flow Cytometry was used to analyze the effect of cycle and apoptosis. Results PDGFR-α shRNA had effect on hRPE cells 24 ～ 72 h, the proliferation of hRPE cells were inhibited, and showed a time-dose response （P 〈 0. 05 ） ; after treated with PDGFR-α shRNA 48 h, the expression of mRNA and protein of PDGFR-α gene was significantly reduced compared with the control group and negative control group（P 〈0. 05） ; the apoptosis rate of experimental group was （37. 10 ±0. 55）%, （50. 8 ±1.41 ）%, compared with the blank control group （11.7 ±1.11 ）% and the negative control group（ 13.5 ±1.05 ）%, the difference was statistically significant （ P 〈0. 05 ） ; the percentage of G1 phase of experimental group was （64.76± 1.16） %, （75.64 ±0. 93）%, compared with the blank control group （54. 51 ±1.40）% and the negative control group （56.04 ±2. 19） %, the difference was statistically significant （ P 〈 0. 05 ）. Conclusion PDGFR-α shRNA can inhibit hRPE cells proliferation and induce apoptosis.

关 键 词：血小板源性生长因子受体-α RNA干扰 视网膜色素上皮细胞 增殖 凋亡 

分 类 号：R774.1[医药卫生—眼科]