Wang Li, Zhu Dan, Yang Yang, He Yuan, Sun Jing, Li Yi-Ming, Wang Zi-Jing, Li Peng
Department of Optometry, Xi'an Medical University, Xi'an 710021, Shaanxi Province, China.
Department of Ophthalmology, the Second Affiliated Hospital of Xi'an Medical University, Xi'an 710038, Shaanxi Province, China.
Int J Ophthalmol. 2024 Oct 18;17(10):1791-1799. doi: 10.18240/ijo.2024.10.04. eCollection 2024.
To explore the DNA methylation of in ultraviolet-B (UVB)-induced age-related cataract (ARC) models and .
Human lens epithelium B3 (HLEB3) cells and Sprague Dawley rats were exposure to UVB respectively. The MTT assay was utilized to evaluate cell proliferation. Flow cytometry was employed for analysis of cell apoptosis and cell cycle. expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction (RT-PCR). The localization of in HLEB3 cells was determined by immunofluorescence. The methylation status of CpG islands located in promoter was verified using bisulfite-sequencing PCR (BSP). DNMTs and TETs mRNA levels was examined by RT-PCR.
UVB exposure decreased HLEB3 cells proliferation, while increased the apoptosis rate and cells were arrested in G0/G1 phase. expression was markedly inhibited in UVB treated cells compared to the controls. Hypermethylation status was detected in the CpG islands within promoter in HLEB3 cells subjected to UVB exposure. Expressions of DNMTs including DNMT1/2/3 were elevated in UVB treated HLEB3 cells compared to that in the controls, while expressions of TETs including TET1/2/3 showed the opposite trend. Results from the UVB treated rat model further confirmed the decreased expression of , hypermethylation status of the CpG islands at promoter of and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.
DNA hypermethylation of promoter CpG islands is correlated with decreased expression in UVB induced HLEB3 cells and anterior lens capsules of rats.
探讨紫外线B（UVB）诱导的年龄相关性白内障（ARC）模型中[具体基因名称未给出]的DNA甲基化情况。
分别将人晶状体上皮B3（HLEB3）细胞和Sprague Dawley大鼠暴露于UVB。采用MTT法评估细胞增殖。运用流式细胞术分析细胞凋亡和细胞周期。使用蛋白质免疫印迹法和逆转录-聚合酶链反应（RT-PCR）评估HLEB3细胞和晶状体前囊膜中[具体基因名称未给出]的表达。通过免疫荧光法确定HLEB3细胞中[具体基因名称未给出]的定位。使用亚硫酸氢盐测序PCR（BSP）验证位于[具体基因名称未给出]启动子区域的CpG岛的甲基化状态。通过RT-PCR检测DNA甲基转移酶（DNMTs）和TET蛋白家族（TETs）的mRNA水平。
UVB照射降低了HLEB3细胞的增殖，同时增加了凋亡率，细胞停滞在G0/G1期。与对照组相比，UVB处理的细胞中[具体基因名称未给出]的表达明显受到抑制。在UVB照射的HLEB3细胞中，检测到[具体基因名称未给出]启动子内的CpG岛存在高甲基化状态。与对照组相比，UVB处理的HLEB3细胞中包括DNMT1/2/3在内的DNMTs表达升高，而包括TET1/2/3在内的TETs表达呈现相反趋势。UVB处理的大鼠模型结果进一步证实了UVB暴露组中[具体基因名称未给出]表达降低、[具体基因名称未给出]启动子CpG岛的高甲基化状态以及DNMT1/2/3和TET1/2的异常表达。
[具体基因名称未给出]启动子CpG岛的DNA高甲基化与UVB诱导的HLEB3细胞和大鼠晶状体前囊膜中[具体基因名称未给出]表达降低相关。
