Detection of „Hotspot Mutations in Catalytic Subunit of Phosphatidylinositol 3-Kinase (Pik3ca) by Allele-Specific Polymerase Chain Reaction

• Published in: Acta medica martiniana Vol. 14; no. 2; pp. 10 - 15
• Main Authors: A, Mendelova, V, Holubekova, P, Zubor, E, Jezkova, K, Gemzova, L, Plank, J, Danko, Z, Lasabova
• Format: Journal Article
• Language: English
• Published: Martin De Gruyter Open 01-09-2014; De Gruyter Poland; Sciendo
• Subjects: allele-specific PCR; Allelopathy; Catalytic oxidation; hotspots; Mutation; PI3K pathway; PIK3CA; Polymerase chain reaction
• ISSN: 1335-8421; 1335-8421; 1338-4139
• DOI: 10.2478/acm-2014-0007

The phosphatidylinositol 3-kinases (PI3Ks) are a family of proteins involved in the regulation of cell survival, growth, metabolism, and glucose homeostasis. Increased PI3K activity is associated with many cancers. PIK3CA gene (encoding p110 , the catalytic subunit of PI3K) is commonly mutated in breast cancer. In our study we focused on the detection of “hotspot” mutations in exons 9 and 20 of the PIK3CA gene in paraffin-embedded tissue of patients with breast cancer. We optimized conditions of allele specific polymerase chain reaction (PCR) and we used direct sequencing to verify our results. Overall, three “hotspot” mutations in PIK3CA gene in paraffin-embadded tissue from breast cancer were detected by allele-specific PCR. All results were verified by direct sequencing of PCR products and we observed 100% agreement between those two methods. We confirmed that allele-specific PCR assay is low cost method usefull for accurate detection of PIK3CA mutations.