Analytic, Preanalytic, and Clinical Validation of p53 IHC for Detection of TP53 Missense Mutation in Prostate Cancer.
Analytic, Preanalytic, and Clinical Validation of p53 IHC for Detection of TP53 Missense Mutation in Prostate Cancer.
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DOI：
10.1158/1078-0432.ccr-17-0257
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发表时间：
2017-08-15
期刊：
影响因子：
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通讯作者：
Lotan TL
中科院分区：
文献类型：
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作者：
Guedes LB;Almutairi F;Haffner MC;Rajoria G;Liu Z;Klimek S;Zoino R;Yousefi K;Sharma R;De Marzo AM;Netto GJ;Isaacs WB;Ross AE;Schaeffer EM;Lotan TL
TP53错义突变可能有助于识别具有致死潜力的前列腺癌。在这里，我们通过预分析、分析和临床验证了一种强大的免疫组织化学（IHC）检测方法，以检测PCa中亚克隆和局灶性TP53错义突变。p53免疫组化检测在clia认可的实验室Ventana Benchmark免疫染色系统上进行。P53蛋白核聚集定义为在10%的肿瘤细胞中存在任何P53核标记。研究了54株来自NCI-60组的福尔马林固定石蜡包埋（FFPE）细胞株和103例已知TP53突变状态的FFPE PCa组织（88例原发性腺癌，15例转移）。DU145和VCaP异种移植物接受不同的固定条件，以研究分析前变量的影响。临床验证在两个部分重叠的根治性前列腺切除术（RP）队列中进行。在NCI-60组中，IHC检测p53核积累对TP53错义突变的灵敏度为100%（25/25错义突变被正确识别）。缺乏p53核积累的特异性为86%(25/29)，因为缺乏TP53错义突变。在FFPE前列腺肿瘤中，p53核积累对潜在错义突变的阳性预测值（PPV）为84%(38/45)，阴性预测值（NPV）为97%（56/58）。在RP术后经历生化复发的男性队列中，与没有p53核积累的患者相比，p53核积累患者发生转移的多变量危险比为2.55 （95% CI: 1.1-5.91）。免疫组化是评估临床PCa标本中是否存在有害和异质TP53错义突变的一种广泛可用的方法。
TP53 missense mutations may help to identify prostate cancer (PCa) with lethal potential. Here, we pre-analytically, analytically and clinically validated a robust immunohistochemistry (IHC) assay to detect subclonal and focal TP53 missense mutations in PCa. The p53 IHC assay was performed in a CLIA-accredited laboratory on the Ventana Benchmark immunostaining system. p53 protein nuclear accumulation was defined as any p53 nuclear labeling in >10% of tumor cells. 54 formalin fixed paraffin embedded (FFPE) cell lines from the NCI-60 panel and 103 FFPE PCa tissues (88 primary adenocarcinomas, 15 metastases) with known TP53 mutation status were studied. DU145 and VCaP xenografts were subjected to varying fixation conditions to investigate the effects of pre-analytic variables. Clinical validation was performed in two partially overlapping radical prostatectomy (RP) cohorts. p53 nuclear accumulation by IHC was 100% sensitive for detection of TP53 missense mutations in the NCI-60 panel (25/25 missense mutations correctly identified). Lack of p53 nuclear accumulation was 86% (25/29) specific for absence of TP53 missense mutation. In FFPE prostate tumors, the positive predictive value (PPV) of p53 nuclear accumulation for underlying missense mutation was 84% (38/45), while the negative predictive value (NPV) was 97% (56/58). In a cohort of men who experienced biochemical recurrence after RP, the multivariable hazard ratio for metastasis among cases with p53 nuclear accumulation compared to those without was 2.55 (95% CI: 1.1–5.91). IHC is widely available method to assess for the presence of deleterious and heterogeneous TP53 missense mutations in clinical PCa specimens.