目的:通过新兴的纳米粒子基因载体观察p21cip1基因对视网膜色素上皮(retinal pigment epithelium,RPE)细胞增殖的抑制情况。方法:制备p21cip1基因纳米粒子,将其转染到培养的人RPE细胞,通过免疫组织化学检测p21cip1蛋白的表达,流式细胞仪检测细胞周期的相关细胞量的变化。结果:p21cip1纳米粒子的DNA含量为3%,包封效率为78%,p21cip1基因在体内由于PLGA-PVA载体的保护作用,可以维持比质粒更长时间的有效期,减少质粒易被生物体内核酸酶降解的问题。流式细胞仪检测显示转染了目的基因的RPE细胞发生G1期阻滞,细胞增殖明显受到抑制。免疫组织化学检测结果显示转染了目的基因的RPE细胞p21cip1蛋白表达明显增强。结论:p21cip1基因有可能作为一个目的基因,借助新兴的基因载体纳米粒子用于抑制细胞增殖的基因治疗。

AIM:To study the inhibition of p21cip1 gene on proliferation of cultured human retinal pigment epithelial(RPE) cells by application of nano-technology.METHODS:Preparation of p21cip1 gene nano-particles was transfected into cultured human RPE cells for detection of expression of p21cip1by immunohistochemistry.Cell cycle related cell volume changes were detected by flow cytometry.RESULTS:DNA content of p21cip1 nano-particles was 3%,encapsulation efficiency was 78%.Because of the protective effects of PLGA-PVA vector p21cip1 gene in the body could maintain a longer valid period than naked plasmid,overcoming problems of the naked plasmid prone to nuclease degradation in vivo.Flow cytometry results showed RPE cells of transfected object gene had G1 phase arrest,significantly inhibited cell proliferation.Immunohistochemical detection results showed RPE cells of transfected object gene had markedly stronger expression of p21cip1.CONCLUSION:p21cip1 gene may serve as a target gene,with the emerging nano-particles of gene vector,to be used for gene therapy in inhibition of cell proliferation.

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