Department of Clinical Genetics Chair of Laboratory and Clinical Genetics, Medical University of Lodz, 92-213 Lodz, Poland.
Department of Otolaryngology, Head and Neck Oncology, Medical University of Lodz, 93-143 Lodz, Poland.
Biomolecules. 2021 May 31;11(6):818. doi: 10.3390/biom11060818.
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: (10-12%), (6%), and (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 H1047R mutation by Real Time PCR (RT-qPCR) and droplet digital PCR (ddPCR). and 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 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 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 mutation assessment in FFPE samples.
头颈部鳞状细胞癌（HNSCC）是第七大常见的人类恶性肿瘤，由于诊断和治疗较晚，其生存率较低。其病因多种多样；然而，遗传因素很重要。HNSCC 中最常见的突变发生在以下基因中：(10-12%)、(6%)和(7-9%)。在某些情况下，这些生物标志物与复发或生存相关，具有潜在的预后和预测价值。共收集了 113 例福尔马林固定石蜡包埋（FFPE）的 HNSCC 肿瘤样本（口腔：35 例（31.0%）；口咽：30 例（26.0%）；喉：48 例（43.0%））。我们通过实时 PCR（RT-qPCR）和液滴数字 PCR（ddPCR）检测 H1047R 突变。通过 RT-qPCR 分析 和 突变，通过免疫组织化学评估 p16 蛋白表达。最后，我们通过 RT-qPCR 确定 HPV 感染。使用 Yates 校正卡方检验或 Fisher 确切检验评估基因组改变与临床参数之间的关系用于名义变量。Kaplan-Meier 图用于生存分析。我们的结果显示，通过 ddPCR 检测到 9 个 H1047R 突变：其中 8 个在 RT-qPCR 中为阴性。由于用于测试 基因突变的方法不同，可能会做出不同的治疗决策。这就是为什么使用最敏感的可用方法非常重要。我们证实了 ddPCR 在 FFPE 样本中 突变评估中的有用性。
