DNA hypermethylation of COL4A1 in ultraviolet-B-induced age-related cataract models in vitro and in vivo.

IF 1.9 4区 医学 Q2 OPHTHALMOLOGY
International journal of ophthalmology Pub Date : 2024-10-18 eCollection Date: 2024-01-01 DOI:10.18240/ijo.2024.10.04
Li Wang, Dan Zhu, Yang Yang, Yuan He, Jing Sun, Yi-Ming Li, Zi-Jing Wang, Peng Li
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引用次数: 0

Abstract

Aim: To explore the DNA methylation of COL4A1 in ultraviolet-B (UVB)-induced age-related cataract (ARC) models in vitro and in vivo.

Methods: 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. COL4A1 expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction (RT-PCR). The localization of COL4A1 in HLEB3 cells was determined by immunofluorescence. The methylation status of CpG islands located in COL4A1 promoter was verified using bisulfite-sequencing PCR (BSP). DNMTs and TETs mRNA levels was examined by RT-PCR.

Results: UVB exposure decreased HLEB3 cells proliferation, while increased the apoptosis rate and cells were arrested in G0/G1 phase. COL4A1 expression was markedly inhibited in UVB treated cells compared to the controls. Hypermethylation status was detected in the CpG islands within COL4A1 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 COL4A1, hypermethylation status of the CpG islands at promoter of COL4A1 and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.

Conclusion: DNA hypermethylation of COL4A1 promoter CpG islands is correlated with decreased COL4A1 expression in UVB induced HLEB3 cells and anterior lens capsules of rats.

紫外线-B 诱导的老年性白内障模型体外和体内 COL4A1 的 DNA 超甲基化。
目的:探讨COL4A1的DNA甲基化在紫外线B(UVB)诱导的老年性白内障(ARC)模型中的体外和体内作用。方法:将人晶状体上皮B3(HLEB3)细胞和Sprague Dawley大鼠分别暴露于紫外线B。采用 MTT 法评估细胞增殖。流式细胞仪用于分析细胞凋亡和细胞周期。利用 Western 印迹和反转录聚合酶链反应(RT-PCR)评估了 COL4A1 在 HLEB3 细胞和晶状体前囊中的表达。免疫荧光法测定了 COL4A1 在 HLEB3 细胞中的定位。利用亚硫酸氢盐测序 PCR(BSP)验证了 COL4A1 启动子中 CpG 岛的甲基化状态。RT-PCR检测了DNMTs和TETs mRNA水平:结果:紫外线照射减少了 HLEB3 细胞的增殖,同时增加了细胞的凋亡率,细胞停滞在 G0/G1 期。与对照组相比,UVB 处理的细胞中 COL4A1 的表达明显受到抑制。在暴露于 UVB 的 HLEB3 细胞中,COL4A1 启动子内的 CpG 岛检测到了高甲基化状态。与对照组相比,经 UVB 处理的 HLEB3 细胞中 DNMTs(包括 DNMT1/2/3)的表达升高,而 TETs(包括 TET1/2/3)的表达则呈相反趋势。UVB处理大鼠模型的结果进一步证实了UVB暴露组中COL4A1的表达下降、COL4A1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:结论:COL4A1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中COL4A1表达的降低有关。
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来源期刊
CiteScore
2.50
自引率
7.10%
发文量
3141
审稿时长
4-8 weeks
期刊介绍: · International Journal of Ophthalmology-IJO (English edition) is a global ophthalmological scientific publication and a peer-reviewed open access periodical (ISSN 2222-3959 print, ISSN 2227-4898 online). This journal is sponsored by Chinese Medical Association Xi’an Branch and obtains guidance and support from WHO and ICO (International Council of Ophthalmology). It has been indexed in SCIE, PubMed, PubMed-Central, Chemical Abstracts, Scopus, EMBASE , and DOAJ. IJO JCR IF in 2017 is 1.166. IJO was established in 2008, with editorial office in Xi’an, China. It is a monthly publication. General Scientific Advisors include Prof. Hugh Taylor (President of ICO); Prof.Bruce Spivey (Immediate Past President of ICO); Prof.Mark Tso (Ex-Vice President of ICO) and Prof.Daiming Fan (Academician and Vice President, Chinese Academy of Engineering. International Scientific Advisors include Prof. Serge Resnikoff (WHO Senior Speciatist for Prevention of blindness), Prof. Chi-Chao Chan (National Eye Institute, USA) and Prof. Richard L Abbott (Ex-President of AAO/PAAO) et al. Honorary Editors-in-Chief: Prof. Li-Xin Xie(Academician of Chinese Academy of Engineering/Honorary President of Chinese Ophthalmological Society); Prof. Dennis Lam (President of APAO) and Prof. Xiao-Xin Li (Ex-President of Chinese Ophthalmological Society). Chief Editor: Prof. Xiu-Wen Hu (President of IJO Press). Editors-in-Chief: Prof. Yan-Nian Hui (Ex-Director, Eye Institute of Chinese PLA) and Prof. George Chiou (Founding chief editor of Journal of Ocular Pharmacology & Therapeutics). Associate Editors-in-Chief include: Prof. Ning-Li Wang (President Elect of APAO); Prof. Ke Yao (President of Chinese Ophthalmological Society) ; Prof.William Smiddy (Bascom Palmer Eye instituteUSA) ; Prof.Joel Schuman (President of Association of University Professors of Ophthalmology,USA); Prof.Yizhi Liu (Vice President of Chinese Ophtlalmology Society); Prof.Yu-Sheng Wang (Director of Eye Institute of Chinese PLA); Prof.Ling-Yun Cheng (Director of Ocular Pharmacology, Shiley Eye Center, USA). IJO accepts contributions in English from all over the world. It includes mainly original articles and review articles, both basic and clinical papers. Instruction is Welcome Contribution is Welcome Citation is Welcome Cooperation organization International Council of Ophthalmology(ICO), PubMed, PMC, American Academy of Ophthalmology, Asia-Pacific, Thomson Reuters, The Charlesworth Group, Crossref,Scopus,Publons, DOAJ etc.
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