PAX6 promoter methylation correlates with MDA-MB-231 cell migration, MMP2 and MMP9 expression.

URRUTIA G; LAURITO SERGIO; CAMPOY EMANUEL; NASIF DANIELA; BRANHAM MT; MARÍA ROQUÉ

ASIAN PACIFIC JOURNAL OF CANCER PREVENTION

ASIAN PACIFIC ORGANIZATION CANCER PREVENTION

Año: 2018

1513-7368

Background: Breast cancer is a heterogeneous disease characterized by the accumulation of genetic and epigenetic alterations, that lead tumor cells to acquire characteristics like invasion and metastasis capacity. Although targeted therapies have improved the management and outcome of patients, metastasis remains being a mayor challenge, since more that 90% of patients diagnosed will die from its cause. Therefore, understanding of the molecular basis of this process, as identification of novel biomarkers can result in improved prevention, diagnosis and treatment of breast cancer patients. Objective: To understand how promoter DNA methylation regulates PAX6 gene expression and influences breast carcinoma cell migration. Methods: PAX6 promoter methylation was evaluated Methyl Specific-Multiplex Ligation Probe Amplification (MS-MLPA). Gene expression was evaluated using qRT-PCR, while effect of PAX6 on migration was evaluated by wound healing assay. In addition, MMP2 and MMP6 genes were studied using different bioinformatic tools. Results: PAX6 promoter is methylated on breast cancer cell lines and methylation in this region impacts on its expression. Migration assays revealed that PAX6 overexpression promotes cell migration, while PAX6 inhibition decreases it. More importantly, we found that migration is affected by PAX6 methylation status. Employing bioinformatic analysis, we found binding sites for PAX6 on the regulatory regions of the MMP2 and MMP9 genes and PAX6 overexpression increases MMP2 and MMP9 expression at the mRNA level. Conclusion: Our study provides novel insights into the epigenetic events that regulate PAX6 expression and molecular mechanisms by which PAX6 modifies the migration capacity of breast cancer cells.