Clinicas Hospital, Faculty of Medicine, State University of São Paulo, Botucatu 18618-682, Brazil.
Department of Pathology and Legal Medicine, Ribeirão Preto School of Medicine, University of São Paulo, Ribeirão Preto 14049-900, Brazil.
Hum Pathol. 2019 Jan;83:177-191. doi: 10.1016/j.humpath.2018.08.026. Epub 2018 Sep 13.
To demonstrate the usefulness of complementary next-generation sequencing (NGS) and immunohistochemistry (IHC) counting, we analyzed 196 patients with non-small cell lung cancer who underwent surgical resection and adjuvant chemotherapy. Formalin-fixed, paraffin-embedded samples of adenocarcinoma (ADC), squamous cell carcinoma, and large cell carcinoma were used to prepare tissue microarrays and were examined by protein H-score IHC image analysis and NGS for oncogenes and proto-oncogenes and genes of tumor suppressors, immune checkpoints, epithelial-mesenchymal transition factors, tyrosine kinase receptors, and vascular endothelial growth factors. In patients with brain metastases, primary tumors expressed lower PIK3CA protein levels. Overexpression of p53 and a higher PD-L1 protein H-score were detected in patients who underwent surgical treatment followed by chemotherapy as compared with those who underwent only surgical treatment The absence of brain metastases was associated with wild-type sequences of genes EGFR, CD267, CTLA-4, and ZEB1. The combination of protein overexpression according to IHC and mutation according to NGS was rare (ie, represented by a very low percentage of concordant cases), except for p53 and vascular endothelial growth factor. Our data suggest that protein levels detected by IHC may be a useful complementary tool when mutations are not detected by NGS and also support the idea to expand this approach beyond ADC to include squamous cell carcinoma and even large cell carcinoma, particularly for patients with unusual clinical characteristics. Conversely, well-pronounced immunogenotypic features seemed to predict the clinical outcome after univariate and multivariate analyses. Patients with a solid ADC subtype and mutated genes EGFR, CTLA4, PDCD1LG2, or ZEB1 complemented with PD-L1 or p53 protein lower expression that only underwent surgical treatment who develop brain metastases may have the worst prognosis.
为了展示互补的下一代测序（NGS）和免疫组织化学（IHC）计数的有用性，我们分析了 196 名接受手术切除和辅助化疗的非小细胞肺癌患者。使用福尔马林固定、石蜡包埋的腺癌（ADC）、鳞状细胞癌和大细胞癌样本制备组织微阵列，并通过蛋白 H 评分 IHC 图像分析和 NGS 检测致癌基因和原癌基因以及肿瘤抑制基因、免疫检查点、上皮-间充质转化因子、酪氨酸激酶受体和血管内皮生长因子。在有脑转移的患者中，原发性肿瘤表达的 PIK3CA 蛋白水平较低。与仅接受手术治疗的患者相比，接受手术治疗加化疗的患者中检测到 p53 过表达和更高的 PD-L1 蛋白 H 评分。没有脑转移与基因 EGFR、CD267、CTLA-4 和 ZEB1 的野生型序列有关。根据 IHC 进行的蛋白过表达和根据 NGS 进行的突变的组合很少（即代表极少数一致性病例），除了 p53 和血管内皮生长因子。我们的数据表明，当 NGS 未检测到突变时，IHC 检测到的蛋白水平可能是一种有用的补充工具，并且还支持将这种方法扩展到 ADC 以外，包括鳞状细胞癌甚至大细胞癌，特别是对于具有不寻常临床特征的患者。相反，明显的免疫基因型特征似乎可以预测单因素和多因素分析后的临床结果。仅接受手术治疗的具有固体 ADC 亚型和突变基因 EGFR、CTLA4、PDCD1LG2 或 ZEB1 并伴有 PD-L1 或 p53 蛋白低表达的患者发生脑转移，可能预后最差。
