LRWD1 expression is regulated through DNA methylation in human testicular embryonal carcinoma cells.

IF 2.4 3区医学 Q2 ANDROLOGY Basic and Clinical Andrology Pub Date : 2021-05-20 DOI:10.1186/s12610-021-00130-y

Jui-Hsiang Hung, Han-Yi Cheng, Yung-Chieh Tsai, Hsien-An Pan, Hany A Omar, Chien-Chih Chiu, Yin-Mei Su, Yung-Ming Lin, Yen-Ni Teng

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引用次数: 2

Abstract

Background: Sperm growth and maturation are correlated with the expression levels of Leucine-rich repeat and WD repeat-containing protein 1 (LRWD1), a widely expressed protein in the human testicles. The decrease in LRWD1 cellular level was linked to the reduction in cell growth and mitosis and the rise in cell microtubule atrophy rates. Since DNA methylation has a major regulatory role in gene expression, this study aimed at exploring the effect of the modulation of DNA methylation on LRWD1 expression levels.

Results: The results revealed the presence of a CpG island up of 298 bps (- 253 ~ + 45) upon LRWD1 promoter in NT2/D1 cells. The hypermethylation of the LRWD1 promoter was linked to a reduction in the transcription activity in NT2/D1 cells, as indicated by luciferase reporter assay. The methylation activator, floxuridine, confirmed the decrease in the LRWD1 promoter transcriptional activity. On the other hand, 5-Aza-2'-deoxycytidine (5-Aza-dc, methylation inhibitor), significantly augmented LRWD1 promoter activity and the expression levels of mRNA and proteins. Furthermore, DNA methylation status of LRWD1 promoter in human sperm genomic DNA samples was analyzed. The results indicated that methylation of LRWD1 promoter was correlated to sperm activity.

Conclusions: Thus, the regulation of LRWD1 expression is correlated with the methylation status of LRWD1 promoter, which played a significant role in the modulation of spermatogenesis, sperm motility, and vitality. Based on these results, the methylation status of LRWD1 promoter may serve as a novel molecular diagnostic marker or a therapeutic target in males' infertility.

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LRWD1在人睾丸胚胎癌细胞中的表达通过DNA甲基化调控。

背景:精子的生长和成熟与人类睾丸中广泛表达的富含亮氨酸重复序列和含有亮氨酸重复序列的蛋白1 (LRWD1)的表达水平有关。LRWD1细胞水平的降低与细胞生长和有丝分裂的减少以及细胞微管萎缩率的上升有关。由于DNA甲基化在基因表达中具有重要的调控作用，本研究旨在探讨DNA甲基化调控对LRWD1表达水平的影响。结果:在NT2/D1细胞中，LRWD1启动子上存在CpG岛，CpG岛长度为- 253 ~ + 45，高298 bps。荧光素酶报告基因实验表明，LRWD1启动子的高甲基化与NT2/D1细胞中转录活性的降低有关。甲基化激活剂氟尿定证实了LRWD1启动子转录活性的降低。另一方面，5-Aza-2′-脱氧胞苷(5-Aza-dc，甲基化抑制剂)显著增强了LRWD1启动子活性以及mRNA和蛋白的表达水平。此外，我们还分析了人类精子基因组DNA样本中LRWD1启动子的DNA甲基化状态。结果表明，LRWD1启动子的甲基化与精子活性相关。结论:LRWD1的表达调控与LRWD1启动子的甲基化状态相关，在精子发生、精子运动和活力的调节中发挥重要作用。基于这些结果，LRWD1启动子的甲基化状态可能作为一种新的分子诊断标记或男性不育症的治疗靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Basic and Clinical Andrology Medicine-Urology

CiteScore

3.50

自引率

0.00%

发文量

审稿时长

22 weeks

期刊介绍： Basic and Clinical Andrology is an open access journal in the domain of andrology covering all aspects of male reproductive and sexual health in both human and animal models. The journal aims to bring to light the various clinical advancements and research developments in andrology from the international community.