title: Detection of EML4-ALK in lung adenocarcinoma using pleural effusion with FISH, IHC, and RT-PCR methods.
abstract: Anaplastic lymphoma kinase (ALK) and echinoderm microtubule-associated protein-like 4 (EML4) gene rearrangements occur in approximately 5% of non-small-cell lung cancers (NSCLC), leading to the overexpression of anaplastic lymphoma kinase and predicting a response to the targeted inhibitor, crizotinib. Malignant pleural effusion occurs in most patients with advanced lung cancer, especially adenocarcinoma, and tissue samples are not always available from these patients. We attempted to clarify the feasibility of detecting the EML4-ALK fusion gene in pleural effusion cells using different methods. We obtained 66 samples of pleural effusion from NSCLC patients. The pleural effusion fluid was centrifuged, and the cellular components obtained were formalin fixed and paraffin embedded. The EML4-ALK fusion gene status was determined with fluorescent in situ hybridization (FISH), reverse transcription-polymerase chain reaction (RT-PCR), and immunohistochemistry (IHC). EML4-ALK was detected in three of 66 patient samples (4.5%) with RT-PCR. When the RT-PCR data were used as the standard, one false positive and one false negative samples were identified with IHC; and one false negative sample was identified with FISH. These results suggest that a block of pleural effusion cells can be used to detect the EML4-ALK fusion gene. IHC had good sensitivity, but low specificity. FISH had low sensitivity, but high specificity. RT-PCR is a good candidate method for detecting EML4-ALK in blocks of pleural effusion cells from lung cancer patients.
authors.affiliation: Department of Pathology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China.
authors.forename: Leilei
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Medicine, Nanjing Thoracic Hospital, Nanjing, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhan
authors.affiliation: Department of Pathology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China.
authors.forename: Xiaodie
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Respiratory Medicine, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of Pathology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Medicine, Nanjing Thoracic Hospital, Nanjing, China.
authors.forename: Like
authors.initials: L
authors.lastname: Yu
authors.affiliation: Department of Pathology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China.
authors.forename: Jiandong
authors.initials: J
authors.lastname: Wang
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology; D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008175:Lung Neoplasms / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D015514:Oncogene Proteins, Fusion / Q000096:biosynthesis*; D016066:Pleural Effusion, Malignant / Q000378:metabolism* / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: C522504:EML4-ALK fusion protein, human; D015514:Oncogene Proteins, Fusion