非小细胞肺癌恶性胸水中癌细胞EML4-ALK重排检测及其临床意义

Detection of EML4-ALK gene rearrangement in NSCLC cells and its clinical significance

摘要:

目的探讨非小细胞肺癌（NSCLC）患者恶性胸水（MPE）中癌细胞EML4-ALK 基因重排的可行性及与其病理特征之间的关系。方法选取138例伴有MPE的NSCLC患者，用瑞氏染色法鉴定MPE 中的癌细胞，用免疫组织化学法鉴定肿瘤组织中EML4-ALK 基因重排，运用微流控芯片技术检测NSCLC 患者MPE 中癌细胞，采用荧光染色法确定癌细胞，采用实时荧光定量聚合酶链反应（qRT-PCR）检测ALK基因重排状态。结果在138 例NSCLC 患者中，肿瘤组织免疫组织化学（IHC）染色有10 例EML4-ALK 基因阳性，qRT-PCR检测到10例EML4-ALK 融合基因重排。IHC和qRT-PCR的一致性为100%，两者敏感性和特异性均为100%。ALK 基因重排状况与患者EGFR突变等差异有统计学意义（P <0.05），而与病理分型、原发部位、临床分期及淋巴结转移等差异无统计学意义（P >0.05）。结论qRT-PCR 检测NSCLC 患者MPE 中癌细胞EML4-ALK 融合基因是筛选基因重排方法的一种补充，值得在临床中推广。

Abstract:

Objective To investigate the application of detecting echinoderm microtubule -associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK ) gene rearrangement by qRT-PCR in non-small cell lung cancer cells (NSCLC), and to evaluate the relationship between EML4-ALK gene rearrangement and clinical relevance. Methods A total of 138 patients confirmed with NSCLC in malignant pleural effusion (MPE) were recruited in this study. Cancer cells in MPE were identified by Wright's staining and microfluidic chip technology. EML4-ALK gene rearrangement in cancer cells was determined by Immunohistochemistry (IHC), and quantitative real time polymerase chain reaction (qRT-PCR). Results Ten out of 138 (7.25%) patients were identified with EML4-ALK rearrangement by IHC, who were also verified by qRT -PCR.Sensitivity and specificity of IHC and qRT -PCR were 100%. The EML4-ALK gene rearrangement was correlated with EGFR mutation (P < 0.05), while no relation between the EML4-ALK gene rearrangement and tumor characters including histological type, site of primary onset, clinical stage and lymphatic metastasis (P >0.05). Conclusions qRT-PCR is a reliable method for detection of EML4-ALK rearrangement detection in NSCLC.