免疫组织化学分析胸苷酸合成酶表达分析的间变性淋巴瘤激酶荧光原位杂交阳性非小细胞肺癌中 3 种不同克隆：一项多中心、回顾性、意大利研究。

Immunohistochemistry with 3 different clones in anaplastic lymphoma kinase fluorescence in situ hybridization positive non-small-cell lung cancer with thymidylate synthase expression analysis: a multicentre, retrospective, Italian study.

机构信息

Department of Oncology, San Luigi Gonzaga Hospital, University of Torino, Orbassano, Italy.

Unit of Pathology, Department of Oncology, Fondazione Poliambulanza Hospital Institute, Brescia, Italy.

出版信息

Pathologica. 2022 Aug;114(4):278-287. doi: 10.32074/1591-951X-756. Epub 2022 Sep 8.

DOI:10.32074/1591-951X-756

PMID:36083243

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9624140/

Abstract

INTRODUCTION

rearrangement is the only druggable oncogenic driver detectable by immunohistochemistry (IHC) not requiring further confirmation of positivity in accessing first-line specific inhibitors. ALK-positive patients experience clinical benefit from pemetrexed-based chemotherapy possibly due to lower thymidylate synthase (TS) levels. This study assesses agreement with three different ALK IHC clones in 37 FISH-positive NSCLC. TS expression by real time (RT)-PCR was compared with FISH-negative cases.

MATERIALS AND METHODS

37 ALK FISH-positive NSCLC cases diagnosed between 2010 and 2015 in 7 Italian centres were investigated with ICH using three different anti-ALK antibodies (ALK1, 5A4 and D5F3). Staining for ALK1 and 5A4 was graded as 0+,1+,2+, and 3+, while the scoring for D5F3 was recorded as negative or positive. Proportion agreement analysis was done using Cohen's unweighted kappa (k). TS and β-actin expression levels were analysed by quantitative RT-PCR. Comparison between TS expression in ALK FISH-positive specimens and a control cohort of ALK FISH-negative ones was performed with the Mann-Whitney and Kruskal-Wallis tests.

RESULTS

Considering 2+ and 3+ as positive, the proportion of IHC agreement was 0.1691 (95% CI 0-0.4595) for ALK1/5A4, 0.1691 (95% CI 0-0.4595) for ALK1/D5F3, and 1 for D5F3/5A4. Considering 3+ as positive, it was 0.1543 (95% CI 0-0.4665) for ALK1/ 5A4, 0.0212 (95% CI 0-0.1736) for ALK1/D5F3, and 0.2269 (95% CI 0-0.5462) for 5A4/D5F3. Median TS expression was 6.07 (1.28-14.94) and ALK-positive cases had a significant lower TS expression than ALK-negative tumours

CONCLUSIONS

IHC proved to be a reliable tool for the diagnosis of ALK-rearranged NSCLC. D5F3 and 5A4 clones have the highest percentage of agreement. TS levels are significantly lower in FISH-positive patients.

摘要

简介

重排是唯一可通过免疫组织化学（IHC）检测到的药物驱动致癌基因，在使用一线特异性抑制剂时不需要进一步确认其阳性。ALK 阳性患者从培美曲塞为基础的化疗中获得临床获益，可能是由于胸苷酸合成酶（TS）水平较低。本研究评估了三种不同 ALK IHC 克隆在 37 例 FISH 阳性 NSCLC 中的一致性。通过实时（RT）-PCR 比较了 TS 的表达与 FISH 阴性病例。

材料和方法

7 家意大利中心在 2010 年至 2015 年间诊断了 37 例 ALK FISH 阳性 NSCLC 病例，使用三种不同的抗 ALK 抗体（ALK1、5A4 和 D5F3）进行 IHC 检测。ALK1 和 5A4 的染色被评为 0+、1+、2+和 3+，而 D5F3 的评分记录为阴性或阳性。使用 Cohen 的未加权 kappa（k）进行比例一致性分析。通过定量 RT-PCR 分析 TS 和 β-肌动蛋白的表达水平。用 Mann-Whitney 和 Kruskal-Wallis 检验比较 ALK FISH 阳性标本和 ALK FISH 阴性对照标本的 TS 表达。

结果

如果将 2+和 3+判定为阳性，则 ALK1/5A4、ALK1/D5F3 的 IHC 一致性比例分别为 0.1691（95%CI 0-0.4595）和 0.1691（95%CI 0-0.4595），D5F3/5A4 为 1。如果将 3+判定为阳性，则 ALK1/5A4、ALK1/D5F3 的一致性比例分别为 0.1543（95%CI 0-0.4665）和 0.0212（95%CI 0-0.1736），D5F3/5A4 为 0.2269（95%CI 0-0.5462）。TS 的中位表达为 6.07（1.28-14.94），ALK 阳性病例的 TS 表达明显低于 ALK 阴性肿瘤。