Zhao Ken, Chen Zhen, Lv Xu-Dong, Dong Gang, Xia Huan
Department of Ophthalmology, The Second Affiliated Hospital of Hubei Polytechnic College; The People's Hospital of Daye City Daye 435100, Hubei Province, P. R. China.
Department of Ophthalmology, Renmin Hospital of Wuhan University Wuhan 430060, Hubei Province, P. R. China.
Am J Transl Res. 2016 Dec 15;8(12):5723-5728. eCollection 2016.
This study aims to explore the impact of micro RNA miR-145 on retinal pigment epithelial cell proliferation and apoptosis.
A stable culture and passage system of hPNE cells was first established, and its migration ability was determined. Then, miR-145 lentiviral vectors were constructed to transfect hPRE cells. Thereafter, hRPE cell proliferation was detected by MTT assay after they were transfected by lentivirus, cell cycle was analyzed by flow cytometry, and apoptosis was detected by Annexin V/PI double staining immunofluorescence.
Cultured hPRE cells had good migrating and metastatic ability, in which subsequent lentivirus infection experiments can be carried out. After transfection by miR-145 lentiviral vectors, hPRE cell proliferation slowed down and RPE cells in the G phase was inhibited; thus, apoptosis rate increased.
MiR-145 can slow down retinal pigment epithelial cell proliferation and increase their apoptosis rate. This has a certain therapeutic potential for diseases caused by RPE cell proliferation such as PVR.
本研究旨在探讨微小RNA miR - 145对视网膜色素上皮细胞增殖和凋亡的影响。
首先建立hPNE细胞稳定的培养和传代体系，并测定其迁移能力。然后构建miR - 145慢病毒载体转染hPRE细胞。之后，慢病毒转染hRPE细胞后，采用MTT法检测细胞增殖，通过流式细胞术分析细胞周期，采用Annexin V/PI双染免疫荧光法检测细胞凋亡。
培养的hPRE细胞具有良好的迁移和转移能力，可在此基础上进行后续的慢病毒感染实验。经miR - 145慢病毒载体转染后，hPRE细胞增殖减缓，G期RPE细胞受到抑制，凋亡率增加。
MiR - 145可减缓视网膜色素上皮细胞增殖并提高其凋亡率。这对增殖性玻璃体视网膜病变（PVR）等由RPE细胞增殖引起的疾病具有一定的治疗潜力。
