Zheng Hong-Hua, Xu Guo-Xing, Guo Jian, Fu Li-Cheng, Yao Yao
Fujian Institute of Ophthalmology, the First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, Fujian Province, China.
Int J Ophthalmol. 2016 Jan 18;9(1):15-20. doi: 10.18240/ijo.2016.01.03. eCollection 2016.
To investigate the role of Aquaporin-1 (AQP-1) in lens epithelial cells (LECs) and its potential target genes. AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance. Herein, AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.
LECs were transfected with lentivirus carrying AQP-1 small interfering RNA (siRNA). Real-time polymerase chain reaction (PCR) and Western blotting were conducted to detect AQP-1 expression in LECs from different groups. Meanwhile, cell counting kit-8 (CCK-8) assay and flow cytometry were performed to measure LEC proliferation and apoptosis, respectively.
AQP-1 expression was significantly reduced in LECs, both at mRNA and protein levels (P<0.05), after siRNA treatment. Decreased cell viability was detected by CCK-8 assay in LECs with siRNA interference, compared to control cells (P<0.05). The apoptosis rate significantly increased in cells after siRNA interference (P<0.05).
The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs. AQP-1 reduction might lead to changes of physiological functions in LECs, which might be associated with the occurrence and development of cataracts.
研究水通道蛋白-1(AQP-1)在晶状体上皮细胞(LECs)中的作用及其潜在的靶基因。AQP-1在眼部晶状体上皮细胞中特异性表达,对晶状体的内环境稳定和透明度维持具有重要意义。在此,研究了AQP-1在晶状体上皮细胞中的表达,以评估其对细胞存活的影响及其在白内障形成中的潜在作用。
用携带AQP-1小干扰RNA(siRNA)的慢病毒转染晶状体上皮细胞。采用实时聚合酶链反应(PCR)和蛋白质免疫印迹法检测不同组晶状体上皮细胞中AQP-1的表达。同时,分别采用细胞计数试剂盒-8(CCK-8)法和流式细胞术检测晶状体上皮细胞的增殖和凋亡情况。
siRNA处理后,晶状体上皮细胞中AQP-1的mRNA和蛋白水平均显著降低(P<0.05)。与对照细胞相比,CCK-8法检测发现siRNA干扰的晶状体上皮细胞活力降低(P<0.05)。siRNA干扰后细胞凋亡率显著增加(P<0.05)。
AQP-1下调后细胞活力降低主要是由于其诱导晶状体上皮细胞凋亡。AQP-1的减少可能导致晶状体上皮细胞生理功能的改变,这可能与白内障的发生和发展有关。