非小细胞肺癌间变性淋巴瘤激酶融合基因的荧光原位杂交检测规范化流程探讨

Standard protocol of ALK fusion gene assessment by fluorescent in situ hybridization in non-small cell lung cancer

目的探讨非小细胞肺癌（NSCLC）患者荧光原位杂交（FISH）方法检测间变性淋巴瘤激酶（ALK）融合基因的操作规范。方法收集中国医学科学肿瘤医院病理科2011年1月至2012年7月共146例NSCLC患者肿瘤病理组织学标本，应用Vysis公司的ALK基因断裂探针、采用FISH方法对ALK融合基因进行检测，荧光显微镜下根据荧光信号进行结果判读。结果146例合格NSCLC患者病理组织学标本，其中手术切除标本110例（75．4％），肺活检标本11例（7．5％），淋巴结转移标本19例（13．0％），其他转移灶6例（4．1％）。ALK融合基因阳性率为8．9％（13／146）。结论采用标准化的流程进行病理组织标本切片、切片预处理、FISH方法检测ALK融合基因是可行的。该流程适用于检测包括手术切除和支气管镜活检标本、淋巴结及其他转移灶标本在内的常见标本ALK融合基因的检测。

Objective To investigate the standard protocol for anaplastic lymphoma kinase （ALK） fusion geue assessment by fluorescent in situ hybridization （FISH） in non-small cell lung cancer （NSCLC）. Methods Tissue specimens of NSCLC cases were retrospectively collected from Jan. 2011 to July 2012. ALK fusion gene was examined by FISH using break-apart ALK gene probes （Vysis company）. The identification of ALK fusion gene was determined by fluorescent signals under a fluorescence microscope. Results One hundred and forty-six eligible NSCLC tumor specimens were tested for ALK fusion gene by FISH. The specimens included 110 cases （75.4%） of surgically-removed tissues, ll cases （7.5%） of biopsy, 19 cases （ 13.0% ） of lymph node and 6 cases （4. 1% ） of other metastatic tissues. The positivity of ALK fusion gene was 8.9% （ 13/146 ）. Conclusions The assessment of ALK fusing gene by FISH using standard protocol for formalin-fixed, paraffin-embedded （FFPE） tissue is feasible. The protocol can used to test in surgically-removed tissues, biopsies, metastatic lymph nodes and other metastastic specimens.