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| 组蛋白甲基化酶SUV39H1过表达对C2C12细胞成骨分化的影响 |
| Effect of histone methylation enzyme SUV39H1 over-expression on osteogenic differentiation of C2C12 cells |
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| DOI：10.3969/j.issn.1006.7108.2022.09.002 |
| 中文关键词: 组蛋白甲基化酶 SUV39H11 C2C12细胞 成骨分化 |
| 英文关键词:histone methylesterase SUV39H1 C2C12 cells osteogenic differentiation |
| 基金项目:国家重点研发计划项目（2018YFC1704300）；国家自然科学基金项目（81973883，81730107）；中国博士后科学基金（2021M692155）；上海市自然科学基金项目（19ZR1458000） |
| 作者 | 单位 | | 王乾1,2 黄晨1,2 黄廷锐1,2 孙攀1,2 赵永见1,2,3 施杞1,2,3 王拥军1,2,3 唐德志1,2,3* | 1.上海中医药大学附属龙华医院, 上海 200032
2.上海中医药大学脊柱病研究所,上海 200032
3.教育部筋骨理论与治法重点实验室,上海 200032 |
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| 中文摘要: |
| 目的 组蛋白甲基化修饰作为组蛋白密码的重要组成部分在表观遗传学研究中占有重要地位。本文旨在探讨组蛋白甲基化酶SUV39H1过表达对骨形态发生蛋白2（bone morphogenetic protein 2，BMP2）诱导的C2C12细胞成骨分化的影响。方法 利用SUV39H1慢病毒感染C2C12细胞构建SUV39H1过表达稳转株。然后利用100 ng/mL的BMP2成骨诱导液对SUV39H1过表达稳转株成骨诱导7 d，随后进行碱性磷酸酶（alkaline phosphatase，ALP）染色和茜素红染色检测成骨分化和矿化相关指标，实时荧光定量PCR(qRT-PCR)、细胞免疫荧光、Western blot检测成骨相关蛋白表达。结果 ALP和茜素红染色显示，与过表达GFP对照组比较，SUV39H1过表达会抑制C2C12细胞早期和晚期成骨分化和矿化程度，qRT-PCR显示，与过表达GFP对照组比较，SUV39H1过表达会降低Runt相关转录因子2（runt-related transcription factor 2，Runx2）、ALP、骨桥蛋白（osteopontin, OPN）、骨钙素（osteocalcin，OCN）基因的表达(P<0.05)。Western blot和免疫荧光结果显示，与过表达GFP对照组比较，SUV39H1过表达会抑制Runx2和ALP成骨蛋白的表达，同时会提高组蛋白H3第9位点赖氨酸三甲基化（histone H3 lysine 9 trimethylation，H3K9me3）蛋白表达水平（P<0.05）。结论 SUV39H1过表达会抑制C2C12细胞的成骨分化水平。 |
| 英文摘要: |
| Objective Histone methylation modifications as an important part of the histone code are important in epigenetic studies. This study aims to investigate the effect of histone methylation enzyme SUV39H1 over-expression on osteogenic differentiation of C2C12 cells induced by bone morphogenetic protein 2 (BMP2). Methods The SUV39H1 over-expression strain was constructed by infecting C2C12 cells with SUV39H1 lentivirus. The osteogenic induction of the SUV39H1 over-expression strain was then performed using 100ng/ml of BMP2 osteogenic induction solution for 7 days. Alkaline phosphatase (ALP) staining and Alizarin red staining were used to detect osteogenic differentiation and mineralization-related indices. Real-time fluorescence quantitative PCR (qRT-PCR), cellular immunofluorescence, and Western blotting were performed to detect osteogenic-related protein expressions. Results ALP and Alizarin red staining results showed that SUV39H1 over-expression inhibited early and late osteogenic differentiation and mineralization in C2C12 cells compared to the GFP over-expression control. qRT-PCR results showed that SUV39H1 over-expression decreased the expression of Runt-related transcription factor 2 (RUNX2), ALP, osteopontin (OPN) and osteocalcin (OCN) genes compared to the GFP over-expression control (P<0.05). Western blotting and immunofluorescence results showed that SUV39H1 over-expression suppressed the expression of Runx2 and ALP osteogenic protein, while increased the expression level of histone H3 lysine9 trimethylation protein compared to the GFP over-expression control (P<0.05). Conclusion Over-expression of histone methylesterase SUV39H1 inhibits osteogenic differentiation of C2C12 cells. |
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