HTRA1基因过表达对视网膜色素上皮细胞增殖及迁移影响

Effects on HTRA1 overexpression on proliferation and migration of RPE cells

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，lit—PCR和WesternBlot检测HTRA1mRNA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，Transwell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。

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 es- tablish 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 trans- duced 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.