蛋白酶体抑制剂对人晶状体上皮细胞的增殖抑制作用

摘要

Objective To investigate the effect of proteasome inhibitor MG132 on the proliferation of human lens epithelial cells SRA01/04. Methods The SRA01/04 cells were treated with MG132 by different concentrations (0, 0. 1, 0. 5, 1. 0, 2. 5, 5.0, 10. 0μmol/L) for 36 hours. The cell viability in all groups was determined using methylthiazoltetrazolium (MTT) test. The effect of MG132 on the apoptosis and regulation of cell cycle about SRA01/04 cells were detected by flow cytometry (FCM). The SRA01/04 cells treated with MG132 were observed after Annexin V/FITC-PI staining by fluorescence microscope. Results The inhibitory effect of MG132 on SRA01/04 cells proliferation was enhanced with the increase of MG132 concentration. The 50% inhibiting concentration ( IC50 ) of MG132 was 2. 50μmol/L after SRA01/04 cells were treated with MG132 for 36 hours. The apoptosis index of the cells treated by MG132 at 2. 5μmol/L and 5 μmol/L for 36 hours was 6. 55 ± 0. 35% and 13.75 ± 3.18%, and 0. 75 ± 0. 21% for 5.0μmol/L for 36 hours in control group. After cells were treated with MG132 for 48h, the percentages of cells at G0/G1 phase were (42. 57 ± 0. 64) %, (73.42 ± 3.10) %, ( 80. 95 ± 3.83 ) % 0, 2. 5,5.0 μmol/Lgroups respectively, and those at S phase were (49. 44±1.36)%, ( 17. 40 ± 1.50)%, ( 19. 57 ± 1.29)%.Annexin V/FITC-PI staining was used, and MG132 was found to result to apoptosis. Conclusions MG132 could inhibit the proliferation of SRA01/04 cells by the effect of inducing apoptosis and regulation of cell cycle. The proteasome inhibitor-might play a key role in the prevention of posterior capsular opacification.%目的 探讨蛋白酶体抑制剂MG132对SRA01/04细胞增殖的影响.方法 以不同浓度MG132处理SRA01/04细胞36 h,通过MTT微量比色法检测MG132对SRA01/04细胞活力的影响,通过流式细胞术(FCM)检测MG132对SRA01/04细胞凋亡及细胞周期的影响,通过AnnexinV/FITC-PI双染法观察MG132对SRA01/04细胞的影响.结果 随着浓度的提高(0、0.1、0.5、1.0、2.5、5.0、10.0 μmol/L),MG132对SRA01/04细胞增殖的抑制作用逐渐增强.36 h时,半数有效抑制浓度IC50为2.50μmol/L.FCM检测细胞凋亡结果:2.5、5.0 μmol/L的MG132作用于SRA01/04细胞36 h,细胞早期凋亡率分别为(6.55±0.35)%和(13.75±3.18)%,而对照组为(0.75±0.21)%,差异有统计学意义(P＜0.01).2.5、5.0 μmol/L MG132处理SAR01/04细胞48 h,G0/G1期细胞比例分别为(73.42±3.10)%,(80.95%±3.83)%,而对照组为(42.57±0.64)%,差异有统计学意义(P＜0.01);S期细胞比例分别为(17.40±1.50)%,(19.57±1.29)%,而对照组为(49.44±1.36)%,差异有统计学意义(P＜0.01).免疫荧光镜下,MG132诱导SRA01/04细胞的早期凋亡.结论 蛋白酶体抑制剂MG132诱导细胞凋亡、影响细胞周期来抑制SRA01/04细胞的增殖.蛋白酶体抑制剂可起到防治后发性白内障的作用.