MLH1 Methylation Status and Microsatellite Instability in Patients with Colorectal Cancer

• Published in: Genes 2025-02, Vol.16 (2), p.182
• Main Authors: Rico-Méndez, Manuel Alejandro, Trujillo-Rojas, Miguel Angel, Ayala-Madrigal, María de la Luz, Hernández-Sandoval, Jesús Arturo, González-Mercado, Anahí, Gutiérrez-Angulo, Melva, Romero-Quintana, José Geovanni, Valenzuela-Pérez, Jesús Alonso, Ramírez-Ramírez, Ruth, Flores-López, Beatriz Armida, Moreno-Ortiz, José Miguel
• Format: Article
• Language: English
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes16020182

Background/Objectives: The purpose of the current study was to compare the methylation of five regions of the CpG island of MLH1 with the presence of microsatellite instability (MSI) in colorectal cancer (CRC) patients. Methods: The study analyzed 138 CRC tumor samples. DNA extraction was performed, followed by bisulfite conversion. MLH1 gene methylation was assessed by methylation-specific PCR (MS-PCR), and the resulting fragments were analyzed using polyacrylamide gels. MSI was evaluated using multiplex PCR, and the fragments were run through capillary electrophoresis. R studio (v4.4.1) and SPSS (v29.0) software were used for the statistical analysis, and values of p < 0.05 were considered statistically significant. Results: The study showed 75.4% unmethylated, 21% partially methylated, and 3.6% fully methylated samples, with region A frequently methylated. MSI was observed in 7.2% of cases (MSI-H: 5.8%, MSI-L: 1.4%). BAT-26 was the most unstable marker. A significant difference between MLH1 methylation and MSI-H (p < 0.01) was identified, but there was no relationship with specific MLH1 regions. Conclusions: No differences were identified when analyzing specific methylation regions in relation to MSI. This study is the first to describe MSI frequency in Mexican patients regardless of age.