Detection of RASSF1A methylation in human prostate cancer using methylation-sensitive high-resolution melting (MS-HRM)

• Main Authors: Cano, Jason Kidd B., Concepcion, Joey Brianne C., Jerez, Gabrielle Rose R., Juridico, Kristine Faith T., Castillo, Josefino R.
• Format: text
• Published: Animo Repository 2019
• Subjects: Prostate—Cancer; Prostate—Diseases—Diagnosis; Methylation; Male Urogenital Diseases
• Online Access: https://animorepository.dlsu.edu.ph/faculty_research/11202
• Institution: De La Salle University

Prostate cancer (PCa) is considered as the second leading cause of death due to cancer in men worldwide and the seventh leading cause of high cancer mortality rate in the Philippines. As other diagnostic methods have shown to be less specific, an epigenetic method called DNA methylation has been the focus of more recent studies. PCa is characterized by aberrant hypermethylation patterns at the CpG islands of the DNA, which are found to be correlated with gene inactivation. Ras-associated domain family member 1 isoform A (RASSF1A), a tumor suppressor gene, was utilized in this study. Thirty (30) formalin-fixed paraffin-embedded (FFPE) prostate cancer samples obtained from University of Santo Tomas Hospital underwent DNA isolation. Extracted DNA were subjected to amplification via polymerase chain reaction and its presence was then confirmed using agarose gel electrophoresis. Subsequently, the DNA isolates were subjected to bisulfite treatment that converted cytosine to uracil, then to MS-HRM and RASSF1A methylation status analysis. RASSF1A was found to have a high sensitivity of 72% and a low specificity of 20%. A secondary test with a low sensitivity/high specificity ratio is recommended to check for the negatives in the cohort.