Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University , Xi'an, Shaanxi 710038, China.
ACS Appl Mater Interfaces. 2015 Jan 14;7(1):359-69. doi: 10.1021/am508122e. Epub 2014 Dec 26.
Somatic mutations in the epidermal growth factor receptor (EGFR) gene are common in patients with lung adenocarcinomas and are associated with sensitivity to the small-molecule tyrosine kinase inhibitors (TKIs). For 10%-50% of the patients who experienced malignant pleural effusion (MPE), pathological diagnosis might rely exclusively on finding lung cancer cells in the MPE. Current methods based on polymerase chain reaction were utilized to test EGFR mutation status of MPE samples, but the accuracy of the test data was very low, resulting in many patients losing the chance of TKIs treatment. Herein, we synthesized the sea-urchin-like Au nanocluster (AuNC) with an average diameter of 92.4 nm, composed of 15-nm nanopricks. By introducing abundant sharp nanopricks, the enhancement factor of AuNC reached at 1.97 × 10(7). After capped with crystal violet (CV), polyethylene glycol, and EGFR mutation specific antibody, the AuNC-EGFR had excellent surface-enhanced Raman scattering (SERS) activity and EGFR mutation targeted recognition capability in lung cancer cells. Characteristic SERS signal at 1617 cm(-1) of CV was linear correlation with the number of H1650 cells, demonstrating the minimum detection limit as 25 cells in a 1-mL suspension. The gold mass in single H1650 cells exposed to AuNC-E746_750 for 2 h ranged from 208.6 pg to 231.4 pg, which approximately corresponded to 56-62 AuNCs per cell. Furthermore, SERS was preclinically utilized to test EGFR mutation status in MPE samples from 35 patients with lung adenocarcinoma. Principal component analysis (PCA) and the support vector machine (SVM) algorithm were constructed for EGFR mutation diagnostic analysis, yielding an overall accuracy of 90.7%. SERS measurement based on sea-urchin-like AuNC was an efficient method for EGFR mutation detection in MPE, and it might show great potential in applications such as predicting gene typing of clinical lung cancer in the near future.
表皮生长因子受体 (EGFR) 基因的体细胞突变在肺腺癌患者中很常见，并且与对小分子酪氨酸激酶抑制剂 (TKI) 的敏感性相关。对于 10%-50%经历恶性胸腔积液 (MPE) 的患者，病理诊断可能完全依赖于在 MPE 中发现肺癌细胞。目前基于聚合酶链反应的方法被用于测试 MPE 样本中的 EGFR 突变状态，但测试数据的准确性非常低，导致许多患者失去 TKI 治疗的机会。在这里，我们合成了平均直径为 92.4nm 的海胆状 Au 纳米团簇 (AuNC)，由 15nm 的纳米锐点组成。通过引入丰富的尖锐纳米锐点，AuNC 的增强因子达到 1.97×10(7)。用结晶紫 (CV)、聚乙二醇和 EGFR 突变特异性抗体封闭后，AuNC-EGFR 在肺癌细胞中具有优异的表面增强拉曼散射 (SERS) 活性和 EGFR 突变靶向识别能力。CV 在 1617cm(-1)处的特征 SERS 信号与 H1650 细胞的数量呈线性相关，表明在 1mL 悬浮液中最低检测限为 25 个细胞。在暴露于 AuNC-E746_750 2 小时的单个 H1650 细胞中，金的质量范围为 208.6pg 至 231.4pg，这大约相当于每个细胞 56-62 个 AuNC。此外，SERS 被用于预临床测试 35 名肺腺癌患者的 MPE 样本中的 EGFR 突变状态。构建了主成分分析 (PCA) 和支持向量机 (SVM) 算法进行 EGFR 突变诊断分析，总准确率为 90.7%。基于海胆状 AuNC 的 SERS 测量是一种有效的 MPE 中 EGFR 突变检测方法，在预测临床肺癌基因分型等方面可能具有很大的应用潜力。
