Effect of RBM20 and MURC digenic heterozygosity variation on the structure and biological characteristics of myocardial cells
10.12007/j.issn.0258-4646.2024.10.004
- VernacularTitle:RBM20和MURC双基因杂合变异对心肌细胞结构和生物学特性的影响
- Author:
Qiaowei LI
1
,
2
,
3
;
Yin YUAN
;
Wenqing ZHU
;
Yanfang YANG
;
Feng HUANG
Author Information
1. 福建医科大学省立临床医学院老年医学科,福州 350001
2. 福州大学附属省立医院老年医学科,福州 350001
3. 福建省临床老年病研究所,福州 350001
- Keywords:
RNA-binding motif protein 20;
muscle-restricted coiled-coil;
digenic heterozygosity variation;
cardiomyopathy
- From:
Journal of China Medical University
2024;53(10):882-892,899
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impact of variations in RNA-binding motif protein 20(RBM20)and muscle-restricted coiled-coil(MURC)digenic heterozygosity variation on the structural and biological characteristics of human cardiomyocyte AC 16(an adult left ventricular myocardial cell line).Methods Cardiomyocyte AC 16 cell lines were constructed with control,negative scramble,wild-type,MURC single-gene mutant,RBM20 single-gene mutant,and RBM20 and MURC digenic mutant groups.The localization of RBM20 and MURC in cardiomyocytes,cell area,cytoskeletal arrangement,cytoskeleton-related proteins,cell polarity,and intracellular calcium concentration were observed using Western blotting,immunofluorescence staining,and reverse transcription polymerase chain reaction.Myocardial apoptosis was detected using flow cytometry.Ki-67 staining and wound healing assays were performed to detect cardiomyo-cyte proliferation and migration,respectively.Results Digenic mutations had a more pronounced impact than single-gene mutations in RBM20 or MURC on the structural and biological characteristics of cardiomyocytes,manifested by increased cell area,upregulated mRNA expression of hypertrophy-related genes,such as myosin heavy chain 7 and alpha-actin,increased cytoskeleton disturbance,decreased flu-orescence intensity of cytoskeletal proteins β-tubulin and Vinculin(all P＜0.01);increased fluorescence intensity of the polarity protein Part 6(P＜0.05);and significantly elevated cardiomyocyte apoptosis rate,decreased proliferative activity,and elevated migration rate and intracellular calcium ion concentration(all P＜0.01).Conclusion The digenic heterozygous variation in RBM20 and MURC may induce changes in the morphological structure and biological characteristics of myocardial cells,including increased cell area,cytoskeleton dis-turbance,cell polarity,increased apoptosis rate and mobility,decreased cell proliferation activity,and calcium processing ability.
