A potential novel role of the R36P mutation in CRYGD in congenial cataract.

IF 1.8 3区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Vision Pub Date : 2024-06-26 eCollection Date: 2024-01-01

Chen Tan, Xueting Yu, Junyi Chen, Xinghuai Sun, Li Wang

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引用次数: 0

Abstract

Purpose: Congenital cataract is an important cause of visual impairment in childhood. Our previous study reported that the c.110G>C (p.R36P) mutation in the γD-crystallin gene (CRYGD) was associated with congenital cataract in a Chinese family. This study aimed to investigate the potential underlying mechanism through which the p.R36P mutation leads to congenital cataract.

Methods: Plasmids encoding wide-type human γD-crystallin and the mutant R36P γD-crystallin were transfected into HEK293T and SRA01/04 cells. Protein expression levels, including total, soluble, and insoluble fractions, were quantified by Western blotting. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to assess the mRNA expression of other crystallin genes. Cell viability and apoptosis were evaluated using the CCK-8 assay and flow cytometry, respectively.

Results: The total protein, especially the soluble fraction, was significantly reduced in the R36P mutant, while the insoluble part remained unaffected. The decrease of soluble R36P γD-crystallin could not be rescued by the proteinase inhibitor MG132. The mRNA expression of the R36P mutation was lower, but other crystallin RNAs were unchanged. Cell viability was slightly decreased (11%, p<0.05), and cell apoptosis was not significantly increased (12%, p=0.31).

Conclusions: The significant decrease in soluble R36P γD-crystallin may represent a novel mechanism underlying congenital cataract caused by CRYGD gene mutation.

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CRYGD 中的 R36P 突变在先天性白内障中的潜在新作用。

目的：先天性白内障是导致儿童视力障碍的重要原因之一。我们之前的研究发现，在一个中国家族中，γD-结晶素基因（CRYGD）的 c.110G>C（p.R36P）突变与先天性白内障有关。本研究旨在探讨 p.R36P 突变导致先天性白内障的潜在机制：方法：将编码宽型人γD-结晶素和突变体 R36P γD-结晶素的质粒转染到 HEK293T 和 SRA01/04 细胞中。蛋白表达水平（包括总、可溶和不可溶部分）通过 Western 印迹法进行量化。定量反转录聚合酶链反应（RT-PCR）用于评估其他晶体蛋白基因的 mRNA 表达。细胞活力和细胞凋亡分别采用 CCK-8 检测法和流式细胞术进行评估：结果：R36P 突变体的总蛋白，尤其是可溶性部分明显减少，而不溶性部分则不受影响。蛋白酶抑制剂 MG132 无法挽救可溶性 R36P γD-crystallin 的减少。R36P 突变的 mRNA 表达量较低，但其他晶体蛋白 RNA 的表达量没有变化。细胞活力略有下降（11%，p结论：可溶性 R36P γD-结晶素的明显减少可能是 CRYGD 基因突变导致先天性白内障的一种新机制。

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

Molecular Vision 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Molecular Vision is a peer-reviewed journal dedicated to the dissemination of research results in molecular biology, cell biology, and the genetics of the visual system (ocular and cortical). Molecular Vision publishes articles presenting original research that has not previously been published and comprehensive articles reviewing the current status of a particular field or topic. Submissions to Molecular Vision are subjected to rigorous peer review. Molecular Vision does NOT publish preprints. For authors, Molecular Vision provides a rapid means of communicating important results. Access to Molecular Vision is free and unrestricted, allowing the widest possible audience for your article. Digital publishing allows you to use color images freely (and without fees). Additionally, you may publish animations, sounds, or other supplementary information that clarifies or supports your article. Each of the authors of an article may also list an electronic mail address (which will be updated upon request) to give interested readers easy access to authors.