Detection of <i>PIK3CA</i> Gene Mutation in Head and Neck Squamous Cell Carcinoma Using Droplet Digital PCR and RT-qPCR

• الحاوية / القاعدة: Biomolecules
• المؤلفون الرئيسيون: Edyta M. Borkowska, Magda Barańska, Magdalena Kowalczyk, Wioletta Pietruszewska
• التنسيق: مقال
• اللغة: الإنجليزية
• منشور في: MDPI AG 2021-05-01
• الموضوعات: head and neck squamous cell carcinoma; <i>PIK3CA</i> gene; human papilloma virus; <i>BRCA</i>; p16
• الوصول للمادة أونلاين: https://www.mdpi.com/2218-273X/11/6/818

Head and neck squamous cell carcinomas (HNSCC) are the seventh cause of human malignancy with low survival rate due to late diagnosis and treatment. Its etiology is diverse; however genetic factors are significant. The most common mutations in HNSCC were found in the genes: <i>PIK3CA</i> (10–12%), <i>BRCA1</i> (6%), and <i>BRCA2</i> (7–9%). In some cases, these biomarkers correlate with recurrences or survival showing a potential of prognostic and predictive value. A total of 113 formalin-fixed paraffin embedded (FFPE) tumor samples were collected from patients with HNSCC (oral cavity: 35 (31.0%); oropharynx: 30 (26.0%); larynx: 48 (43.0%)). We examined <i>PIK3CA</i> H1047R mutation by Real Time PCR (RT-qPCR) and droplet digital PCR (ddPCR). <i>BRCA1</i> and <i>BRCA2</i> mutations were analyzed by RT-qPCR while p16 protein expression was assessed by immunohistochemistry. Finally, we identified HPV infection by RT-qPCR. The relationships between genomic alterations and clinical parameters were assessed using the Yates’ corrected Chi-squared test or Fisher’s exact test for nominal variables. Kaplan Meier plots were applied for survival analysis. Our results revealed 9 <i>PIK3CA</i> H1047R mutations detected by ddPCR: 8 of them were negative in RT-qPCR. Due to the use of different methods to test the presence of the <i>PIK3CA</i> gene mutation, different treatment decisions might be made. That is why it is so important to use the most sensitive methods available. We confirmed the usefulness of ddPCR in the <i>PIK3CA</i> mutation assessment in FFPE samples.