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Research article
Yongmin Xiong
Xi'an Jiaotong University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-3351-2128
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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, including 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 GPX3-8_CpG_11, GPX3-8_CpG_16, GPX3-8_CpG_20 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 GPX3-8_CpG_24 in KBD patients with degree III was significantly higher than those in KBD patients with degree I/II (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 logarithmic increase in mRNA levels of GPX3, GPX1 and GPX4 decreased the rate of apoptosis in chondrocytes.
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 development of KBD.
Keywords
Kashin-Beck Disease, GPX3, DNA methylation, CpG
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A new preprint design is coming!
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research article
Yongmin Xiong
Xi'an Jiaotong University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-3351-2128
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
View the published article at Journal of Trace Elements in Medicine and Biology.
Version 1
Posted 13 Jan, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
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License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Journal of Trace Elements in Medicine and Biology.
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, including 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 GPX3-8_CpG_11, GPX3-8_CpG_16, GPX3-8_CpG_20 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 GPX3-8_CpG_24 in KBD patients with degree III was significantly higher than those in KBD patients with degree I/II (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 logarithmic increase in mRNA levels of GPX3, GPX1 and GPX4 decreased the rate of apoptosis in chondrocytes.
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 development of KBD.
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