Investigation of simultaneous of CADM1 gene methylation pattern and HPV virus quantification as a diagnostic biomarker in Cervical Cancer

Abstract

Background and Aim: Researches have shown that promoter methylation of the CADM1 gene is one of the main mechanisms is to silence its expression in cancer cells. The objective of this study was to determine the methylation pattern of the CADM1 gene promoter and to quantify the human papillomavirus. Materials and Methods: In this research, 75 tumor tissues and 75 tumor marginal tissue samples taken from patients with definitive diagnosis of cervical cancer were examined. After extraction of DNA and ensuring the quality and quantity of DNA extracted by spectrophotometry and the agarose gel electrophoresis method, the methylation pattern of the CADM1was performed by the HRM method. Determining the methylation pattern of the CADM1 gene, without considering the type of involved variants was performed by using the Eva Green method and comparing the high-resolution melting point and HPV quantities were determined after isolation from the block. Results: Methylation of CADM1 gene in cervical tumor cells increased significantly (P< 0.0001) in comparison with the tumor margin before and after radiotherapy. But the change in the level of methylation of tumor cells before and after radiotherapy was not significant. This means that radiotherapy does not affect the level of methylation of the CADM1 gene. Moreover, the level of methylation of CADM1 gene in patients with HPV titer higher than 500 ml / IU was significantly more than that of patients with HPV titer lower than 500 ml / IU. Conclusion: Use of DNA methylated of the cancer cells as a prognosis for cervical cancer in HPV positive women it’s appropriate, also the CADM1 gene can be used as a marker of cervical cancer.