大骨节病患者GPX3启动子的CpG甲基化可能促进软骨细胞凋亡。

Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements Pub Date : 2021-01-13 DOI:10.21203/rs.3.rs-142170/v1

Rongqiang Zhang, Di Zhang, XiaoLi Yang, Dandan Zhang, Qiang Li, Chen Wang, Xuena Yang, Y. Xiong

{"title":"大骨节病患者GPX3启动子的CpG甲基化可能促进软骨细胞凋亡。","authors":"Rongqiang Zhang, Di Zhang, XiaoLi Yang, Dandan Zhang, Qiang Li, Chen Wang, Xuena Yang, Y. Xiong","doi":"10.21203/rs.3.rs-142170/v1","DOIUrl":null,"url":null,"abstract":"OBJECTIVE\nTo determine the methylation levels of CpGs in the GPX3 promoter region and explore their potential effects on the apoptosis of chondrocytes.\n\n\nMETHODS\nBlood specimens were collected from 32 participants; 16 KBD patients and 16 healthy subjects. Twenty-five CpGs in the promoter region of GPX3 were identified and detected by MALDI-TOF-MS. Methylation levels of CpGs were compared between KBD patients and healthy subjects as well as among the KBD patients with different degrees. C28/I2 human chondrocytes were treated with tBHP and Na2SeO3. Apoptosis in chondrocytes was examined under a fluorescence microscope.\n\n\nRESULTS\nThe methylation levels of GPX3-1_CpG_11 and GPX3-1_CpG_16 in KBD patients were significantly higher than those of healthy subjects (P < 0.05). The methylation levels of the other CpGs were not significantly different between the two groups (P > 0.05). The methylation level of GPX3-1_CpG_24 in KBD patients was significantly higher than those of healthy subjects (P < 0.05). MSP-PCR analysis indicated that the methylation rate of KBD group (9.41%) was significantly higher than that of healthy subjects (1.18%), and that GPX3 DNA methylation increased the risk of acquiring KBD 8 fold (OR = 8.000, 95% CI: 1.023-62.580); The mRNA expression of GPX3 in whole blood of KBD patients was lower than that of healthy subjects (P＜0.05); Compared with the control group, GPX3, GPX1 and GPX4 mRNA level of the tertbutyl hydroperoxide injury group decreased significantly (P < 0.05), after supplementation with Na2SeO3. The rate of chondrocytes apoptosis was decreased with the increasing of GPX3 and GPX4 mRNA levels (P＜0.05) and GPX3 mRNA showed a similar trend without statistically significant (P＞0.05).\n\n\nCONCLUSION\nThe methylation patterns of CpGs in GPX3 varied in KBD patients. The experiments indicated that the increased methylation of CpGs within the GPX3 promoter may down-regulate the expression of GPX3, thereby reducing the antioxidant function of GPX3 and promoting chondrocyte apoptosis, both of which accelerates the occurrence of KBD. We therefore propose a new understanding of GPX3's potential epigenetic and genetic mechanisms that contribute to KBD.","PeriodicalId":17536,"journal":{"name":"Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements","volume":"3 4","pages":"126943"},"PeriodicalIF":0.0000,"publicationDate":"2021-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"CpG methylation of the GPX3 promoter in patients with Kashin-Beck Disease potentially promotes chondrocyte apoptosis.\",\"authors\":\"Rongqiang Zhang, Di Zhang, XiaoLi Yang, Dandan Zhang, Qiang Li, Chen Wang, Xuena Yang, Y. Xiong\",\"doi\":\"10.21203/rs.3.rs-142170/v1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"OBJECTIVE\\nTo determine the methylation levels of CpGs in the GPX3 promoter region and explore their potential effects on the apoptosis of chondrocytes.\\n\\n\\nMETHODS\\nBlood specimens were collected from 32 participants; 16 KBD patients and 16 healthy subjects. Twenty-five CpGs in the promoter region of GPX3 were identified and detected by MALDI-TOF-MS. Methylation levels of CpGs were compared between KBD patients and healthy subjects as well as among the KBD patients with different degrees. C28/I2 human chondrocytes were treated with tBHP and Na2SeO3. Apoptosis in chondrocytes was examined under a fluorescence microscope.\\n\\n\\nRESULTS\\nThe methylation levels of GPX3-1_CpG_11 and GPX3-1_CpG_16 in KBD patients were significantly higher than those of healthy subjects (P < 0.05). The methylation levels of the other CpGs were not significantly different between the two groups (P > 0.05). The methylation level of GPX3-1_CpG_24 in KBD patients was significantly higher than those of healthy subjects (P < 0.05). MSP-PCR analysis indicated that the methylation rate of KBD group (9.41%) was significantly higher than that of healthy subjects (1.18%), and that GPX3 DNA methylation increased the risk of acquiring KBD 8 fold (OR = 8.000, 95% CI: 1.023-62.580); The mRNA expression of GPX3 in whole blood of KBD patients was lower than that of healthy subjects (P＜0.05); Compared with the control group, GPX3, GPX1 and GPX4 mRNA level of the tertbutyl hydroperoxide injury group decreased significantly (P < 0.05), after supplementation with Na2SeO3. The rate of chondrocytes apoptosis was decreased with the increasing of GPX3 and GPX4 mRNA levels (P＜0.05) and GPX3 mRNA showed a similar trend without statistically significant (P＞0.05).\\n\\n\\nCONCLUSION\\nThe methylation patterns of CpGs in GPX3 varied in KBD patients. The experiments indicated that the increased methylation of CpGs within the GPX3 promoter may down-regulate the expression of GPX3, thereby reducing the antioxidant function of GPX3 and promoting chondrocyte apoptosis, both of which accelerates the occurrence of KBD. We therefore propose a new understanding of GPX3's potential epigenetic and genetic mechanisms that contribute to KBD.\",\"PeriodicalId\":17536,\"journal\":{\"name\":\"Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements\",\"volume\":\"3 4\",\"pages\":\"126943\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2021-01-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.21203/rs.3.rs-142170/v1\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.21203/rs.3.rs-142170/v1","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}

引用次数: 1

摘要

目的检测GPX3启动子区CpGs的甲基化水平，探讨其对软骨细胞凋亡的潜在影响。方法采集32例患者血液标本;16例大骨节病患者和16例健康人。利用MALDI-TOF-MS对GPX3启动子区的25个CpGs进行了鉴定和检测。比较大骨病患者与健康人群以及不同程度大骨病患者的CpGs甲基化水平。用三必和必和必加Na2SeO3处理C28/I2人软骨细胞。荧光显微镜下观察软骨细胞凋亡。结果大肠癌患者GPX3-1_CpG_11和GPX3-1_CpG_16的甲基化水平显著高于健康人(P < 0.05)。大肠癌患者GPX3-1_CpG_24甲基化水平显著高于健康人群(P < 0.05)。MSP-PCR分析显示，大骨病组甲基化率(9.41%)显著高于健康组(1.18%)，GPX3 DNA甲基化使患大骨病的风险增加8倍(OR = 8.000, 95% CI: 1.023 ~ 62.580);大肠癌患者全血GPX3 mRNA表达量低于健康人(P<0.05);与对照组相比，添加Na2SeO3后，叔丁基过氧化氢损伤组GPX3、GPX1和GPX4 mRNA水平显著降低(P < 0.05)。软骨细胞凋亡率随GPX3、GPX4 mRNA表达水平的升高而降低(P<0.05)， GPX3 mRNA表达水平升高趋势相似，但无统计学意义(P>0.05)。结论大肠癌患者GPX3中CpGs的甲基化模式存在差异。实验表明，GPX3启动子内CpGs甲基化增加可能下调GPX3的表达，从而降低GPX3的抗氧化功能，促进软骨细胞凋亡，从而加速大骨节病的发生。因此，我们提出了对GPX3潜在的表观遗传和遗传机制的新理解，这些机制有助于大骨节病。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

CpG methylation of the GPX3 promoter in patients with Kashin-Beck Disease potentially promotes chondrocyte apoptosis.

OBJECTIVE To determine the methylation levels of CpGs in the GPX3 promoter region and explore their potential effects on the apoptosis of chondrocytes. METHODS Blood specimens were collected from 32 participants; 16 KBD patients and 16 healthy subjects. Twenty-five CpGs in the promoter region of GPX3 were identified and detected by MALDI-TOF-MS. Methylation levels of CpGs were compared between KBD patients and healthy subjects as well as among the KBD patients with different degrees. C28/I2 human chondrocytes were treated with tBHP and Na2SeO3. Apoptosis in chondrocytes was examined under a fluorescence microscope. RESULTS The methylation levels of GPX3-1_CpG_11 and GPX3-1_CpG_16 in KBD patients were significantly higher than those of healthy subjects (P < 0.05). The methylation levels of the other CpGs were not significantly different between the two groups (P > 0.05). The methylation level of GPX3-1_CpG_24 in KBD patients was significantly higher than those of healthy subjects (P < 0.05). MSP-PCR analysis indicated that the methylation rate of KBD group (9.41%) was significantly higher than that of healthy subjects (1.18%), and that GPX3 DNA methylation increased the risk of acquiring KBD 8 fold (OR = 8.000, 95% CI: 1.023-62.580); The mRNA expression of GPX3 in whole blood of KBD patients was lower than that of healthy subjects (P＜0.05); Compared with the control group, GPX3, GPX1 and GPX4 mRNA level of the tertbutyl hydroperoxide injury group decreased significantly (P < 0.05), after supplementation with Na2SeO3. The rate of chondrocytes apoptosis was decreased with the increasing of GPX3 and GPX4 mRNA levels (P＜0.05) and GPX3 mRNA showed a similar trend without statistically significant (P＞0.05). CONCLUSION The methylation patterns of CpGs in GPX3 varied in KBD patients. The experiments indicated that the increased methylation of CpGs within the GPX3 promoter may down-regulate the expression of GPX3, thereby reducing the antioxidant function of GPX3 and promoting chondrocyte apoptosis, both of which accelerates the occurrence of KBD. We therefore propose a new understanding of GPX3's potential epigenetic and genetic mechanisms that contribute to KBD.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements

自引率

0.00%

发文量