CAJ | 학술논문

目的研究HTRA1基因过表达对人视网膜色素上皮细胞(RPE)的增殖和迁移能力的影响,探讨HTRA1基因在老年黄斑变性(AMD)发病中的作用.方法用携带人HTRA1基因的慢病毒感染体外培养的人RPE细胞,荧光显微镜下观察细胞的感染效率,RT-PCR和Westem Blot检测HTRAlmRNA和蛋白的表达,MTT比色法检测感染细胞毒性及细胞增殖能力,TransweH小室检测细胞迁移能力变化.结果与空载体慢病毒感染RPE细胞相比,HTRA1基因慢病毒感染的RPE细胞HTRA1mRNA和蛋白的表达量明显增加,细胞增殖能力及迁移能力均明显下降,差异有统计学意义(P＜0.01).结论过表达HTRA1基因能抑制人视网膜色素上皮细胞的增殖及迁移,THRA1基因可能通过影响RPE细胞功能参与AMD的进程.
목적 연구HTRA1기인과표체대인시망막색소상피세포(RPE)적증식화천이능력적영향,탐토HTRA1기인재노년황반변성(AMD)발병중적작용.방법 용휴대인HTRA1기인적만병독감염체외배양적인RPE세포,형광현미경하관찰세포적감염효솔,RT-PCR화Westem Blot검측HTRAlmRNA화단백적표체,MTT비색법검측감염세포독성급세포증식능력,TransweH소실검측세포천이능력변화.결과 여공재체만병독감염RPE세포상비,HTRA1기인만병독감염적RPE세포HTRA1mRNA화단백적표체량명현증가,세포증식능력급천이능력균명현하강,차이유통계학의의(P＜0.01).결론 과표체HTRA1기인능억제인시망막색소상피세포적증식급천이,THRA1기인가능통과영향RPE세포공능삼여AMD적진정.
Objective To study the effects of HTRA1 gene overexpression on proliferative and migration ability of human retinal pigment epithelium (RPE) cells,and investigate the possible way of HTRA1 acting on age-related macular degeneration (AMD). Methods RPE cells was transduced of Lentivirus containing pGC-FU-HTRA1 plasmid and inspected under fluorescent microscopy to establish the infection efficiency.MTT and Transwell assay were performed to detect the proliferation and migration of RPE cells respectively. Results As compared with negative control lentivirus transduced RPE cells,the proliferation and migration of RPE cells overexpressing HTRA1 gene were significantly inhibited. Conclusions Overexpression of HTRA1 gene can inhibit proliferation and migration of RPE cells in vitro,indicating that RPE cells is a possible target for HTRA1 promoting the initiation and development of AMD.