Paolo Bironzo, Laura Melocchi, Valentina Monica, Dario Trebeschi, Fausto Barbieri, Evaristo Maiello, Maria Rita Migliorino, Alessandra Lombardi, Marcello Tiseo, Luisella Righi, Paolo Graziano, Giulio Rossi, Silvia Novello
{"title":"3个不同克隆间变性淋巴瘤激酶荧光原位杂交阳性非小细胞肺癌胸苷合酶表达分析的免疫组织化学:一项多中心，回顾性，意大利研究。","authors":"Paolo Bironzo, Laura Melocchi, Valentina Monica, Dario Trebeschi, Fausto Barbieri, Evaristo Maiello, Maria Rita Migliorino, Alessandra Lombardi, Marcello Tiseo, Luisella Righi, Paolo Graziano, Giulio Rossi, Silvia Novello","doi":"10.32074/1591-951X-756","DOIUrl":null,"url":null,"abstract":"<p><strong>Introduction: </strong><i>ALK</i> 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 <i>ALK</i> FISH-negative cases.</p><p><strong>Materials and methods: </strong>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.</p><p><strong>Results: </strong>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 <i>(p = 0.002).</i></p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":45893,"journal":{"name":"PATHOLOGICA","volume":null,"pages":null},"PeriodicalIF":4.4000,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/39/8c/pathol-2022-04-278.PMC9624140.pdf","citationCount":"0","resultStr":"{\"title\":\"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.\",\"authors\":\"Paolo Bironzo, Laura Melocchi, Valentina Monica, Dario Trebeschi, Fausto Barbieri, Evaristo Maiello, Maria Rita Migliorino, Alessandra Lombardi, Marcello Tiseo, Luisella Righi, Paolo Graziano, Giulio Rossi, Silvia Novello\",\"doi\":\"10.32074/1591-951X-756\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Introduction: </strong><i>ALK</i> 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 <i>ALK</i> FISH-negative cases.</p><p><strong>Materials and methods: </strong>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.</p><p><strong>Results: </strong>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 <i>(p = 0.002).</i></p><p><strong>Conclusions: </strong>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.</p>\",\"PeriodicalId\":45893,\"journal\":{\"name\":\"PATHOLOGICA\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.4000,\"publicationDate\":\"2022-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/39/8c/pathol-2022-04-278.PMC9624140.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"PATHOLOGICA\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.32074/1591-951X-756\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/9/8 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q1\",\"JCRName\":\"PATHOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"PATHOLOGICA","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.32074/1591-951X-756","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/9/8 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"PATHOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
ALK重排是免疫组化(IHC)检测到的唯一可用药的致癌驱动因素，在获得一线特异性抑制剂时无需进一步确认阳性。alk阳性患者从培美曲塞为基础的化疗中获益，可能是由于胸腺苷酸合成酶(TS)水平较低。本研究在37例fish阳性NSCLC中评估了三种不同的ALK IHC克隆的一致性。实时(RT)-PCR法比较ALK - fish阴性患者的TS表达情况。材料和方法:2010年至2015年间，意大利7个中心诊断的37例ALK fish阳性非小细胞肺癌患者使用三种不同的抗ALK抗体(ALK1, 5A4和D5F3)进行ICH调查。ALK1和5A4染色分为0+、1+、2+和3+，D5F3评分为阴性或阳性。采用Cohen's未加权kappa (k)进行比例一致性分析。采用定量RT-PCR分析TS和β-actin表达水平。采用Mann-Whitney和Kruskal-Wallis试验比较ALK fish阳性标本和ALK fish阴性对照标本中TS的表达。结果:考虑2+和3+为阳性，ALK1/ 5a4的IHC一致性比例为0.1691 (95% CI 0-0.4595)， ALK1/D5F3为0.1691 (95% CI 0-0.4595)， D5F3/ 5a4为1。考虑到3+为阳性，ALK1/ 5A4为0.1543 (95% CI 0-0.4665)， ALK1/D5F3为0.0212 (95% CI 0-0.1736)， 5A4/D5F3为0.2269 (95% CI 0-0.5462)。TS的中位表达量为6.07 (1.28-14.94)，alk阳性患者的TS表达量显著低于alk阴性患者(p = 0.002)。结论:免疫组化是诊断alk重排非小细胞肺癌的可靠工具。D5F3和5A4克隆的一致性百分比最高。fish阳性患者的TS水平显著降低。
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.
Introduction: ALK 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 ALK 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 (p = 0.002).
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.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
