Y Zhai1, Y-Y Zhu. 1. Health Management Division, The People's Hospital of Weifang, Weifang, Shandong, China. 13853650797@163.com.
Abstract
OBJECTIVE: To investigate both the relationship and underlying mechanism between miR-30a and migraine. PATIENTS AND METHODS: Peripheral blood samples were collected from migraine patients and healthy people to extract RNA for quantitative Real-time PCR (qRT-PCR). The relationship between mRNA expressions and patient data was analyzed by t-test. Target genes of miR-30a were predicted by the TargetScan. The binding of miR-30a to the target gene was verified by dual fluorescein reporter assay. The protein expression of calcitonin/alpha-CGRP gene (CALCA), the miR-30a target gene, was detected by Western blot. RESULTS: Expression levels of miR-30a in peripheral blood of migraine patients were significantly lower than those of healthy controls detected by qRT-PCR, and the methylation level of miR-30a in promoter region was remarkably increased. In addition, expression levels of miR-30a were significantly decreased in patients with bilateral seizures, persistent pain and high pain index. CALCA was found to be the target gene of miR-30a via bioinformatics analysis. We verified that miR-30a degrades CALCA by dual-luciferase reporter assay. Western blot results showed that overexpression of miR-30a down-regulated the CALCA expression, and knockdown of miR-30a upregulated the CALCA expression. CONCLUSIONS: Expression levels of miR-30a are significantly decreased in migraine patients and can relieve migraine through the degradation of CALCA.
OBJECTIVE: To investigate both the relationship and underlying mechanism between miR-30a and migraine. PATIENTS AND METHODS: Peripheral blood samples were collected from migrainepatients and healthy people to extract RNA for quantitative Real-time PCR (qRT-PCR). The relationship between mRNA expressions and patient data was analyzed by t-test. Target genes of miR-30a were predicted by the TargetScan. The binding of miR-30a to the target gene was verified by dual fluorescein reporter assay. The protein expression of calcitonin/alpha-CGRP gene (CALCA), the miR-30a target gene, was detected by Western blot. RESULTS: Expression levels of miR-30a in peripheral blood of migrainepatients were significantly lower than those of healthy controls detected by qRT-PCR, and the methylation level of miR-30a in promoter region was remarkably increased. In addition, expression levels of miR-30a were significantly decreased in patients with bilateral seizures, persistent pain and high pain index. CALCA was found to be the target gene of miR-30a via bioinformatics analysis. We verified that miR-30a degrades CALCA by dual-luciferase reporter assay. Western blot results showed that overexpression of miR-30a down-regulated the CALCA expression, and knockdown of miR-30a upregulated the CALCA expression. CONCLUSIONS: Expression levels of miR-30a are significantly decreased in migrainepatients and can relieve migraine through the degradation of CALCA.