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首页> 外文期刊>Scientific reports. >Antibody validation and scoring guidelines for ABCG2 immunohistochemical staining in formalin-fixed paraffin-embedded colon cancer tissue
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Antibody validation and scoring guidelines for ABCG2 immunohistochemical staining in formalin-fixed paraffin-embedded colon cancer tissue

机译：福尔马林固定石蜡包埋的结肠癌组织中ABCG2免疫组织化学染色的抗体验证和评分指南

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Overexpression of the ATP-dependent drug efflux pump ABCG2 is a major molecular mechanism of multidrug resistance in cancer and might be a predictive biomarker for drug response. Contradictory results have been reported for immunohistochemical studies of ABCG2 protein expression in colorectal cancer (CRC), probably because of the use of different antibodies and scoring approaches. In this study, we systematically studied six commercially available anti-ABCG2 antibodies, using cell lines with up-regulation of ABCG2, and selected one antibody for validation in CRC tissue. Furthermore, we established scoring guidelines for ABCG2 expression based on the clinically used guidelines for HER2 immunohistochemistry assessment in gastric cancer. The guidelines provide a semi-quantitative measure of the basolateral membrane staining of ABCG2 and disregard the apical membrane staining and the cytoplasmic signal. Intra-tumor heterogeneity in ABCG2 immunoreactivity was observed; however, statistical analyses of tissue microarrays (TMAs) and the corresponding whole sections from primary tumors of 57 metastatic CRC patients revealed a strong positive correlation between maximum TMA scores and whole sections, especially when more than one core was used. In conclusion, here, we provide validated results to guide future studies on the associations between ABCG2 immunoreactivity in tumor cells and the benefits of chemotherapeutic treatment in patients with CRC.
机译：ATP依赖性药物外排泵ABCG2的过表达是癌症多药耐药性的主要分子机制，可能是药物反应的预测生物标志物。已针对大肠癌（CRC）中ABCG2蛋白表达的免疫组织化学研究报告了相反的结果，可能是因为使用了不同的抗体和评分方法。在这项研究中，我们使用具有ABCG2上调的细胞系系统地研究了六种市售抗ABCG2抗体，并选择了一种抗体用于CRC组织中的验证。此外，我们根据临床使用的胃癌HER2免疫组化评估指南，建立了ABCG2表达评分指南。该指南提供了ABCG2基底外侧膜染色的半定量测量，而忽略了顶膜染色和细胞质信号。观察到ABCG2免疫反应性的肿瘤内异质性;然而，对57例转移性CRC患者的原发性肿瘤组织微阵列（TMA）和相应的整个切片的统计分析显示，最大TMA得分与整个切片之间存在很强的正相关性，尤其是在使用多个核的情况下。总之，在这里，我们提供经过验证的结果，以指导今后关于肿瘤细胞中ABCG2免疫反应性与CRC患者进行化学治疗的益处之间的相关性研究。

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