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Phosphatidylinositol-3-kinases (PI3K) are essential for cell signaling,proliferation, differentiation and survival. The catalytic subunit of PI3K, encodedby the PIK3CA oncogene, is mutated in 18-45% of breast carcinomas. These mutations,involved in tumorigenic processes, activate the PI3K/AKT/mTOR signaling pathway.Resistance to anti‑human epidermal growth factor receptor, hormonal or anti-PI3Ktherapies have been described in breast carcinomas bearing activation of the PI3Ksignaling pathway. The present study reports the evaluation of PIK3CA exon 9 and20 mutations in 149 invasive breast cancer cases using a validated PCR-high resolutionmelting assay (PCR-HRM). An amplification refractory mutation system (PCR-ARMS)using allele-specific scorpion primers was used to detect hotspot mutations inexons 9 (c.1624G→A and c.1633G→A) and 20 (c.3140A→G and c.3140A→T) in 118 tumorspecimens. No correlation was observed with age at diagnosis, histological type,hormone receptor and HER2 status. PIK3CA exon 9 and 20 mutations were found tobe related to Scarff-Bloom-Richardson (SBR) grade with a lower rate of mutationsand a higher frequency of exon 9 mutations in SBRI and exon 20 mutations in SBRII/IIItumors. No difference was observed in the incidence rates of the two differentmutations screened for each exon in any subcategory. A statistically significantcorrelation was found between PCR-HRM and PCR-ARMS (κ=0.845; P&lt;0.001). PCR-ARMSwas found to be more sensitive than PCR-HRM (sensitivity 0.5 and 5-10% of mutatedDNA, respectively). We propose that PCR-HRM and PCR-ARMS can be combined for thecost-effective routine clinical identification of PIK3CA mutations for the purposeof personalizing therapy for invasive breast cancers.

Analysis of PIK3CA exon 9 and 20 mutations in breast cancers using PCR-HRM and PCR-ARMS: Correlation with clinicopathological criteria