摘要
目的探讨微小RNA-7(miRNA-7)对人视网膜色素上皮(human retinal pigment epithelium,hRPE)细胞增生和迁移的调控作用。方法利用脂质体Lipofectamine 2000将miRNA-7前体miRNA-145(miRNA-7前体干预组)和阴性对照物(阴性miRNA对照组)转染hRPE细胞,同时设立空脂质体组和空白对照组,采用Real-Time PCR检测成熟miRNA-7的表达,CCK-8检测细胞增殖抑制情况,用直径为8.5 mm的Boyden小室进行细胞迁移能力的检测,对4组结果作统计学分析。结果 miRNA-7对hRPE细胞增殖有明显抑制作用,miRNA-7前体干预组细胞增殖抑制率(23.58%)明显高于阴性miRNA对照组(2.35%)、空脂质体组(2.97%)与空白对照组(0%),差异均具有统计学意义(均为P<0.05),空脂质体组、阴性miRNA对照组与空白对照组之间差异均无统计学意义(均为P>0.05)。在miRNA-7前体干预组中,迁移细胞数明显较少,每高倍视野为(16.0±3.8)个,而在阴性miRNA对照组、空脂质体组和空白对照组中,迁移细胞数明显较多,每高倍视野分别为(32.2±7.8)个、(31.4±6.9)个和(33.9±8.4)个,miRNA-7前体干预组细胞迁移数较后3组显著减少,差异均有统计学意义(均为P<0.01),而后3组之间差异均无统计学意义(均为P>0.05)。结论 miRNA-7可明显抑制hRPE细胞的增殖和迁移,这种抑制作用可能是由于miRNA-7对hRPE细胞的EGFR通路产生了良好的靶向沉默后效应,直接下调EGFR表达,从而阻断或部分阻断了ERK1/2信号通路,使得hRPE细胞的增殖和迁移受到明显抑制。
Objective To evaluate the regulation function of microRNA-7 on pro- liferation and migration of human retinal pigment epithelial （ hRPE ） cells. Methods Using liposomes Lipofectamine2000 transmit microRNA-7 precursor （transfection group） and negative control （ control group） into hRPE cells, then the expression of ma- ture microRNA-7 was detected by RT-PCR, cell proliferative inhibition was detected by CCK 8, and cell migration ability was investigated with a diameter of 8.5 mm Boyden chamber. Results In the transfection group, blank control group, empty liposome group and negative control group, hRPE cell proliferation was significantly inhibited by microRNA-7, the proliferative inhibition rate of transfection group （23.58%）was signifi- cantly higher than that of negative control group （2. 35% ）, empty liposome group （2. 97% ）and blank control group （0%）, there were statistical differences between these groups （ all P 〈 0.05 ）, no statistical difference between empty liposome group, negative control group and blank control group （ all P 〉 0.05 ）. In microRNA-7 transfec- tion group, microRNA-7 inhibited RPE cell migration obviously, compared with the other three groups the cells of migration （ 16.0 ±3.8 ） were significantly reduced （ all P 〈0. 01 ）. In the negative control group, empty liposome group and blank control group, the number of migrating cells were 32.2 ±7.8,31.4 ±5.9 and 5.9 ±8.4, respectively, and there was no statistical difference between three groups （ all P 〉 0. 05）. Conclusion MicroRNA-7 can inhibit the proliferation and migration of hRPE cells obviously, which may be the gene silencing after-effects targeting EGFR pathway （ RPE cells response gene） to reduce EGFR expression directly with block or partially blocked ERK1/2 signaling pathways.
出处
《眼科新进展》
CAS
北大核心
2014年第2期122-126,共5页
Recent Advances in Ophthalmology
基金
广州市番禺区科技计划项目(编号:2011-Z-03-23)~~
