骨关节炎软骨细胞中hsa-miR-3202的生物信息分析与功能验证  

Bioinformatics analysis and functional verification of hsa-miR-3202 in osteoarthritic chondrocytes

作　　者：张佳琪 刘艳虹 梁慧婷 周晶晶 王雅雯 许憬瑜 李煜爽 雷立健 胡晓琴 Zhang Jiaqi;Liu Yanhong;Liang Huiting;Zhou Jingjing;Wang Yawen;Xu Jingyu;Li Yushuang;Lei Lijian;Hu Xiaoqin(Department of Epidemiology,School of Public Health,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学公共卫生学院流行病学教研室,山西省太原市030001

出　　处：《中国组织工程研究》2025年第12期2458-2465,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81803326),项目负责人:胡晓琴;山西省华晋骨科公益基金会资助项目(20211120),项目负责人:梁慧婷;山西省华晋骨科公益基金会资助项目(20221116),项目负责人:周晶晶;山西省应用基础研究项目(201801D221265),项目负责人:胡晓琴;山西省高等学校教学改革创新项目(J20220372),项目负责人:胡晓琴。

摘　　要：背景:软骨细胞增殖与凋亡的失衡在骨关节炎发生发展中起着重要作用,已有研究发现hsa-miR-3202参与调节多种细胞的增殖和凋亡过程,然而尚未有研究探讨hsa-miR-3202与骨关节炎的相关性。目的:探讨hsa-miR-3202在骨关节炎软骨细胞中的表达及其对软骨细胞增殖与凋亡能力的影响。方法:①通过生物信息分析筛选骨关节炎软骨细胞中差异表达的microRNA,基于国内外研究现状,选择hsa-miR-3202进行后续研究,采用GO功能富集和KEGG通路富集对其进行靶基因预测。②选择对数生长期的人正常软骨细胞系C28/I2,随机分4组培养:正常组加入普通培养基培养24 h,更换普通培养基培养6 h,更换普通培养基继续培养;脂多糖组加入含脂多糖的培养基培养24 h,更换普通培养基培养6 h,更换普通培养基继续培养;脂多糖+NC组加入含脂多糖的培养基培养24 h,加入has-miR-3202 mimics对照转染6 h,更换普通培养基继续培养;脂多糖+hsa-miR-3202 mimics组加入含脂多糖的培养基培养24 h,加入has-miR-3202 mimics转染6 h,更换普通培养基继续培养。继续培养48 h后,采用RT-qPCR检测细胞hsa-miR-3202表达,流式细胞术检测细胞凋亡;继续培养0-72 h,采用CCK-8检测细胞增殖活力。结果与结论:①生物信息分析结果显示,hsa-miR-3202表达在骨关节炎软骨细胞中显著下调;GO功能富集与KEGG通路富集结果显示,hsa-miR-3202靶基因功能与细胞生长、凋亡密切相关。②体外细胞实验结果显示,与正常组比较,脂多糖组软骨细胞hsa-miR-3202表达与细胞增殖能力下降(P<0.05),细胞凋亡率增加(P<0.05);与脂多糖组比较,脂多糖+hsa-miR-3202 mimics组软骨细胞hsa-miR-3202表达与细胞增殖能力升高(P<0.05),细胞凋亡率减少(P<0.05)。③结果表明,hsa-miR-3202在骨关节炎软骨细胞中表达下调,其表达下调可抑制细胞增殖并促进细胞凋亡,进而影响骨关节炎疾病的发生和发展。BACKGROUND:The imbalance between proliferation and apoptosis of chondrocytes plays an important role in the occurrence and development of osteoarthritis.Previous studies have found that hsa-miR-3202 is involved in regulating the proliferation and apoptosis of various cells.However,no studies have explored the correlation between hsa-miR-3202 and osteoarthritis.OBJECTIVE:To investigate the expression of hsa-miR-3202 in osteoarthritic chondrocytes and its effect on the proliferation and apoptosis of chondrocytes.METHODS:(1)MicroRNAs differentially expressed in osteoarthritic chondrocytes were screened by biogenic analysis.Based on the current research situation at home and abroad,hsa-miR-3202 was selected for follow-up studies,and its target genes were predicted by gene ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.(2)Human normal chondrocyte cell lines C28/I2 in logarithmic growth phase were selected and randomly divided into four groups for culture:in normal group,cells were cultured in normal medium for 24 hours,the medium was then changed to normal medium for another 6 hours of culture,and changed to normal medium for subsequent culture;in lipopolysaccharide group,cells were cultured in lipopolysaccharide-containing medium for 24 hours,the medium was then changed to normal medium for another 6 hours,and changed to normal medium for subsequent culture;in lipopolysaccharide+NC group,cells were cultured in lipopolysaccharide-containing medium for 24 hours,and then transfected with has-miR-3202 mimics control for 6 hours,and the medium was change to normal medium for subsequent culture;in lipopolysaccharide+hsa-miR-3202 mimics group,cells were cultured in lipopolysaccharide-containing medium for 24 hours and then transfected with has-miR-3202 mimics for 6 hours,and the medium was changed to normal medium for subsequent culture.After further 48 hours of culture,the expression level of hsa-miR-3202 was detected by fluorescence quantitative PCR and cell apoptosis

关 键 词：has-miR-3202 骨关节炎 生物信息学 软骨细胞 炎症 细胞增殖 细胞凋亡 老年人 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R684.3