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Vol. 16. Issue 3.
Pages 229-234 (May - June 2020)
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Vol. 16. Issue 3.
Pages 229-234 (May - June 2020)
Original Article
DOI: 10.1016/j.reumae.2018.06.010
Methylation Status of Interleukin-6 Gene Promoter in Patients with Behçet's Disease
Estatus de la metilación del promotor del gen interleucina-6 en pacientes con síndrome de Behçet
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Shahriar Alipoura,b, Ebrahim Sakhiniac, Alireza Khabbazib, Nasser Samadid, Zohreh Babalooe, Mahdi Azadf, Somayeh Abolhasanig, Jafar Farhadia,b, Golamreza Jadideslama,b, Neda Roshanravanh, Mohammad Nouric,
Corresponding author
nourimd@yahoo.com

Corresponding author.
a Molecular Medicine, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Iran
b Connective Tissue Diseases Research Center, Tabriz University of Medical Science, Iran
c Department of Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
d Cancer Biochemistry, Cancer Biotechnology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
e Department of Immunology Medicine Faculty, Tabriz University of Medical Sciences, Tabriz, Iran
f Department of Medical Laboratory Sciences, Faculty of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, Iran
g Faculty of Dentistry, Tabriz University of Medical Sciences, Tabriz, Iran
h Cardiovascular Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
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Table 1. PCR primers and product size.
Table 2. Clinical profile of patients with IL-6 gene expression and its methylation.
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Abstract
Background

IL-6 mRNA expression is significantly high in many autoimmune diseases such as Behçet's disease; this is often related with more aggressive phenotypes. Nevertheless, the essential molecular process for its high expression has not been completely realized. The aim of this study was undertaken to estimate the gene copy number variation and promoter methylation to IL-6's high expression.

Methods

This study was performed on 51 patients and 61 healthy controls. Initially, DNA and RNA were extracted from all specimens. Promoter methylation levels of IL-6 were evaluated by MeDIP-qPCR technique. Also, IL-6 gene expression was measured by Real-time PCR. After that, we evaluated the relationship between gene expression and methylation, as well as their relationship with clinical specification.

Results

As we expected, the expression level of IL-6 gene increased significantly in the patient group compared to the healthy subjects. Also, the relative promoter methylation level of the IL-6 mRNA was significantly lower in patient group compared to healthy group (p<0.001).

Discussion

We disclosed that the promoter hypomethylation may be considered as one of the main defects for IL-6 mRNA high expression in patients with Behçet's disease.

Keywords:
Behçet's disease
IL-6
MeDIP-qPCR
Methylation
Resumen
Antecedentes

La expresión de ARNm de IL-6 es significativamente elevada en muchas enfermedades autoinmunes, tales como el síndrome de Behçet, y ello se relaciona a menudo con fenotipos más agresivos. Sin embargo, no se ha comprendido plenamente el proceso molecular esencial para esta expresión elevada. El objetivo de este estudio fue la estimación de la variación del número de copias del gen, y la metilación del promotor de la expresión elevada de IL-6.

Métodos

Este estudio se realizó en 51 pacientes y 61 controles sanos. Al inicio, se extrajo ADN y ARN de todas las muestras. Se evaluaron los niveles de metilación del promotor de IL-6 mediante la técnica MeDIP-qPCR. También se midió la expresión del gen IL-6 mediante PCR a tiempo real. Tras ello, evaluamos la relación entre la expresión del gen y la metilación, así como su relación con la especificación clínica.

Resultados

Según lo previsto, el nivel de expresión del gen IL-6 se incrementó significativamente en el grupo de pacientes, con respecto a los sujetos sanos. También encontramos que el nivel relativo de metilación del promotor de ARNm de IL-6 fue considerablemente menor en el grupo de pacientes, con respecto al grupo sano (p<0,001).

Discusión

Concluimos que la hipometilación del promotor puede considerarse uno de los defectos principales de la expresión elevada de ARNm de IL-6, en los pacientes con síndrome de Behçet.

Palabras clave:
Síndrome de Behçet
IL-6
MeDIP-qPCR
Metilación

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