TGF-β1基因转染可抑制人晶状体上皮细胞增殖.docVIP

TGF-β1基因转染可抑制人晶状体上皮细胞增殖 【摘要】 　　目的：探讨脂质体介导的TGF-β1基因(pEGFP-TGF-β1)转染对人晶状体上皮细胞(HLEC)系B3(HLEC- B3)增殖、凋亡及细胞周期的 影响 。 方法 ：将pEGFP-TGF-β1转染HLEC-B3,观察细胞形态变化;采用生长曲线法、噻唑蓝比色法 分析 细胞增殖改变;使用流式细胞仪测定细胞凋亡变化、分析细胞周期改变(DNA倍体法)。结果：pEGFP-TGF-β1成功转染后，HLEC-B3逐渐变圆、脱壁。转染后24～48h细胞的增殖受到抑制,与相应对照组比较，差异均有统计学意义(P 0.01)；凋亡细胞比例(转染24h为（21.0±1.7） %，转染48h至（43.6±1.4）%明显升高,与相应对照组[24h为（0.42±0.06）%，48h为（0.60±0.02%)]比较,差异均有统计学意义(P 0.01)；细胞出现G1期阻滞现象,表现为G1期细胞比例[转染24h为（72.0±1.9 ）%，转染48h为（ 74.7±2.2）%]增加和S期细胞比例[转染24h为（20.4±2.2）%，转染48h为（19.4±1.4）%]下降,与相应对照组比较，差异均有统计学意义(P 0.01)。结论：pEGFP-TGF-β1可成功转染HLEC-B3,诱发细胞G1期阻滞,有效抑制细胞增殖,诱导细胞凋亡。 【关键词】 晶状体上皮细胞　TGF-β1基因　转染　凋亡 　　Inhibitory effect of TGF-β1 gene transfection on the proliferation of human lens epithelial cell Abstract AIM:To investigate the effects of pEGFP-TGF-β1 plasmid DNA on proliferation and survival of human lens epithelial cell. METHODS:Immortalized human lens epithelial cell line(HLEC- B3) was transfected by pEGFP-TGF-β1 genes, respectively. Cell proliferation was analyzed by cell growth curve and MTT colorimetric assay, cell apoptosis and cell cycle of HLEC-B3 were examined by flowcytometry. RESULTS:HLEC-B3 cells were transfected successfully, pEGFP-TGF-β1 suppressed the growth rate, enhanced the cell apoptosis(21.0±1.7) % after 24 hours and (43.6±1.4 )% after 48 hours transfection. The proportions of G1 phase cells were increased to (72.0±1.9)% after 24 hours and (74.7±2.2)% after 48 hours transfection, while the proportions of S phase cells were decreased to(20.5±2.2)% after 24 hours and (19.4±1.4)% after 48 hours transfection. CONCLUSION: pEGFP-TGF-β1 genes can successfully transfect HLEC-B3, inhibit proliferation and induce apoptosis of HLEC-B3. · KEYWORDS: lens epithelial cells;　TGF-β1 genes; transfection; apoptosis 　　0引言 转化生长因子TGF-β1可以参与并调节多种生物学反应过程,其中包括:细胞的生长与分化、细胞粘连及细胞外基质的形成、伤口的愈合、免疫调节及胚胎发育过程等。许多 研究 表明,高表达活性TGF-β1的转基因鼠和其体外培养鼠晶状体均出现晶状体浑浊的症状[1,2]，实验证明TGF-β1可抑制许多正常细胞和肿瘤细胞的生长,并使这些细胞停滞在细胞周期的第一个间隔区:G1期,从而阻止细胞进入S期[3]。本研究的目的是通过 应用 基因转染技术将外源性TGF-β1基因转染至体