手工免疫组化法检测ALK融合蛋白在非小细胞肺癌中的表达

Manual immunohistochemistry for detecting ALK gene fusion antibody in 519 cases of nonsmall lung cancer

目的探讨手工免疫组织化学法（IHC）检测间变性淋巴瘤激酶（ALK）融合蛋白在非小细胞肺癌（NSCLC）中的表达及其意义。方法手工IHC检测519例NSCLC中ALK融合蛋白表达,分析ALK融合蛋白与临床特点、病理特征的关系,比较IHC检测手术与活检标本的差异,探究手工IHC与全自动免疫组化机器（Vantana）法、荧光原位杂交（FISH）的一致性。结果 ALK融合蛋白阳性率为11.37%（59/519）,病人较年轻（P=0.048）,以腺癌为主,多为黏液性腺癌亚型（P〈0.01）,ALK阳性是发生淋巴结转移的危险因素[OR=2.188（95%C.I∶1.161-4.122）];IHC检测ALK融合蛋白与组织大小无关（P〉0.05）;手工IHC强阳性病例与Vantana IHC、FISH有一致性。结论手工IHC法能够成为条件不足的基层医院筛查ALK重排的可靠手段。

Objective To investigate the expression of anaplastic lymphoma kinase（ALK） gene fusion antibody in non-small cell lung cancer（NSCLC） and explore the clinicopathological significance. Methods Using manual immunohistochemistry（IHC）with D5F3 rabbit monoclonal antibody, we detected the expression of ALK gene fusion protein in 519 cases of NSCLC. The relations of ALK fusion protein with the clinical characteristics of the patients and the histological classification of the tumors were analyzed. The expressions of ALK fusion protein were compared between surgical specimens and biopsy samples, and the consistency of manual IHC results was evaluated with the results of a fully automated IHC instrument and fluorescence in situ hybridization（FISH）. Results The positivity rate of ALK fusion protein was 11.37%（59/519） among the cases detected by manual IHC. The patients tended to have a young age of onset（P=0.048） and most of the tumors were adenocarcinoma. In the surgical specimens, ALK fusion protein was expressed mostly in invasive mucinous adenocarcinoma（P〈0.01）, and it was a high risk factor of lymph node metastasis [OR=2.188（95%C.I：1.161-4.122）]. No statistical difference was found in the test results of manual IHC between surgical specimens and biopsy samples. The results by manual IHC suggesting a strong expression were consistent with the results by automated IHC and FISH. Conclusion Manual IHC can be reliable for screening ALK fusion arrangement in patients with NSCLC.