Abstract：
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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).
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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).
UNASSIGNED: 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：
未经证实：ALK重排是通过免疫组织化学（IHC）检测到的唯一可成药的致癌驱动因素，不需要进一步确认获得一线特异性抑制剂的阳性。ALK阳性患者从基于培美曲塞的化疗中获得临床益处，可能是由于较低的胸苷酸合成酶（TS）水平。该研究评估了在37个FISH阳性NSCLC中与三种不同的ALKIHC克隆的一致性。通过实时（RT）-PCR将TS表达与ALKFISH阴性病例进行比较。
UNASSIGNED：使用三种不同的抗ALK抗体（ALK1，5A4和D5F3），对2010年至2015年在7个意大利中心诊断的37例ALKFISH阳性NSCLC病例进行了ICH调查。ALK1和5A4染色分级为0+，1+,2+,3+,而D5F3的评分记录为阴性或阳性。比例一致性分析使用科恩的未加权卡帕(K)进行。通过定量RT-PCR分析TS和β-肌动蛋白表达水平。用Mann-Whitney和Kruskal-Wallis试验对ALKFISH阳性标本和ALKFISH阴性标本中的TS表达进行了比较。
未经评估：考虑2+和3+为正，ALK1/5A4的IHC一致性比例为0.1691(95%CI0-0.4595),ALK1/D5F3为0.1691(95%CI0-0.4595),D5F3/5A4为1.考虑到3+是积极的，ALK1/5A4为0.1543（95%CI0-0.4665），ALK1/D5F3为0.0212（95%CI0-0.1736），5A4/D5F3为0.2269（95%CI0-0.5462）。TS表达中位数为6.07（1.28-14.94），ALK阳性病例的TS表达明显低于ALK阴性肿瘤（p=0.002）。
UNASSIGNED：IHC被证明是诊断ALK重排的NSCLC的可靠工具。D5F3和5A4克隆具有最高的一致性百分比。FISH阳性患者的TS水平明显较低。
UNASSIGNED：使用三种不同的抗ALK抗体（ALK1，5A4和D5F3），对2010年至2015年在7个意大利中心诊断的37例ALKFISH阳性NSCLC病例进行了ICH调查。ALK1和5A4染色分级为0+，1+,2+,3+,而D5F3的评分记录为阴性或阳性。比例一致性分析使用科恩的未加权卡帕(K)进行。通过定量RT-PCR分析TS和β-肌动蛋白表达水平。用Mann-Whitney和Kruskal-Wallis试验对ALKFISH阳性标本和ALKFISH阴性标本中的TS表达进行了比较。
未经评估：考虑2+和3+为正，ALK1/5A4的IHC一致性比例为0.1691(95%CI0-0.4595),ALK1/D5F3为0.1691(95%CI0-0.4595),D5F3/5A4为1.考虑到3+是积极的，ALK1/5A4为0.1543（95%CI0-0.4665），ALK1/D5F3为0.0212（95%CI0-0.1736），5A4/D5F3为0.2269（95%CI0-0.5462）。TS表达中位数为6.07（1.28-14.94），ALK阳性病例的TS表达明显低于ALK阴性肿瘤（p=0.002）。
UNASSIGNED：IHC被证明是诊断ALK重排的NSCLC的可靠工具。D5F3和5A4克隆具有最高的一致性百分比。FISH阳性患者的TS水平明显较低。
