<abstract xmlns="http://www.w3.org/1999/xhtml"><p> 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.</p></abstract>

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.

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

Department of Obstetrics and Gynecology, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Slovak Republic

Department of Molecular Biology, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Slovak Republic

Mgr., Department of Molecular Biology, Jessenius Faculty of Medicine, Comenius University, Martin, Kalinciaka 2, 038 61 Vrutky, Slovakia. Phone: 00421-43-4286517

Department of Pathological Anatomy, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Slovak Republic

Acta Medica Martiniana

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

The phosphatidylinositol 3-kinases (PI3Ks) are a family of proteins involved in the regulation of cell survival, growth, metabolism, and glucose homeostasis....

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

Publié en ligne: 03 oct. 2014

Pages: 10 - 15

Reçu: 22 avr. 2014

Accepté: 23 mai 2014

DOI: https://doi.org/10.2478/acm-2014-0007

Mots clés
PIK3CA, “hotspots”, allele-specific PCR, PI3K pathway

© by Mendelova A.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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

Publié en ligne: 03 oct. 2014

Pages: 10 - 15

Reçu: 22 avr. 2014

Accepté: 23 mai 2014

DOI: https://doi.org/10.2478/acm-2014-0007

Mots clésPIK3CA, “hotspots”, allele-specific PCR, PI3K pathway

© by Mendelova A.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Mots clés
PIK3CA, “hotspots”, allele-specific PCR, PI3K pathway