mRNA Expression and DNA Methylation of CXCL16 in Menstrual Blood and Endometrium Tissue of Subjects with Endometriosis and Pelvic Pain
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This study found increased mRNA expression and hypomethylation of CXCL16 in the menstrual blood of endometriosis patients compared to controls, suggesting it could be a diagnostic marker.
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This study investigated whether CXCL16 mRNA expression and CXCL16 DNA methylation differ between endometriosis and control groups by measuring both in menstrual blood and endometrium tissue from 35 women using qPCR for mRNA and pyrosequencing after bisulfite conversion for DNA methylation. CXCL16 mRNA expression was 2.42-fold higher in menstrual blood of subjects with endometriosis than in controls, and CXCL16 expression in menstrual blood was not significantly different from that in matched endometrial tissue. DNA methylation of CXCL16 was lower in menstrual blood of endometriosis subjects, indicating hypomethylation, but the authors note the need for validation and larger cohort studies to support CXCL16 as a diagnostic marker. This paper is centrally about endometriosis — it assesses CXCL16 mRNA expression and DNA methylation in menstrual blood and endometrium tissue as potential indicators of endometriosis-associated inflammation and pain.
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Abstract
BACKGROUND: The cytokine chemokine ligand 16 (CXCL16) plays an important role in the pathophysiology of endometriosis by regulating the inflammatory response and contributing to the pain-associated endometriosis. Despite this, the impact of epigenetic modifications, such as DNA methylation, on CXCL16 has yet to be fully understood. Therefore, this research was conducted to assess both the mRNA expression and DNA methylation levels of the proinflammatory gene CXCL16 in the endometrium tissue and menstrual blood of patients with and without endometriosis.METHODS: Thirty-five women with and without endometriosis were involved in this research. Subjects' menstrual blood samples were collected using filter paper pads, meanwhile the endometrium tissue were collected by performing biopsy, from which DNA and RNA were extracted. The DNA methylation levels of the CXCL16 were measured using the pyrosequencing method following bisulfite conversion treatment. Meanwhile, the mRNA expression level was measured using the quantitative polymerase chain reaction (qPCR) method and analyzed with the Livak method.RESULTS: The mRNA expression of CXCL16 in menstrual blood of endometriosis subjects was 2.42 times higher compared to control group (p=0.030). Furthermore, the expression of CXCL16 in menstrual blood was identical to that in endometrial tissue (p=0.173). DNA methylation analysis showed that CXCL16 in the menstrual blood of endometriosis subjctes had lower methylation levels compared to controls (p=0.004), indicating hypomethylation.CONCLUSION: Increased mRNA expression and hypomethylation of CXCL16 in the menstrual blood of endometriosis patients could serve as a direct marker for diagnosing endometriosis. However, further study to validate these findings and explore the potential of CXCL16 as a diagnostic tool, and additional research involving larger patient for the cohorts study is necessary.KEYWORDS: CXCL16, DNA methylation, endometrium, menstrual blood, mRNA expression, pain
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Abstract
BACKGROUND: The cytokine chemokine ligand 16 (CXCL16) plays an important role in the pathophysiology of endometriosis by regulating the inflammatory response and contributing to the pain-associated endometriosis. Despite this, the impact of epigenetic modifications, such as DNA methylation, on CXCL16 has yet to be fully understood. Therefore, this research was conducted to assess both the mRNA expression and DNA methylation levels of the proinflammatory gene CXCL16 in the endometrium tissue and menstrual blood of patients with and without endometriosis.
Methods
Thirty-five women with and without endometriosis were involved in this research. Subjects' menstrual blood samples were collected using filter paper pads, meanwhile the endometrium tissue were collected by performing biopsy, from which DNA and RNA were extracted. The DNA methylation levels of the CXCL16 were measured using the pyrosequencing method following bisulfite conversion treatment. Meanwhile, the mRNA expression level was measured using the quantitative polymerase chain reaction (qPCR) method and analyzed with the Livak method.
Results
The mRNA expression of CXCL16 in menstrual blood of endometriosis subjects was 2.42 times higher compared to control group (p=0.030). Furthermore, the expression of CXCL16 in menstrual blood was identical to that in endometrial tissue (p=0.173). DNA methylation analysis showed that CXCL16 in the menstrual blood of endometriosis subjctes had lower methylation levels compared to controls (p=0.004), indicating hypomethylation.
Conclusion
Increased mRNA expression and hypomethylation of CXCL16 in the menstrual blood of endometriosis patients could serve as a direct marker for diagnosing endometriosis. However, further study to validate these findings and explore the potential of CXCL16 as a diagnostic tool, and additional research involving larger patient for the cohorts study is necessary.
Keywords
CXCL16, DNA methylation, endometrium, menstrual blood, mRNA expression, pain
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DOI: https://doi.org/10.18585/inabj.v16i2.2958
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