Smith David Hersi, Fiehn Anne-Marie Kanstrup, Fogh Louise, Christensen Ib Jarle, Hansen Tine Plato, Stenvang Jan, Nielsen Hans Jørgen, Nielsen Kirsten Vang, Hasselby Jane Preuss, Brünner Nils, Jensen Sussie Steen
1] R&D, Dako A/S, Produktionsvej 42, DK-2600 Glostrup, Denmark [2] Section for Molecular Disease Biology, Institute of Veterinary Disease Biology, Faculty of Health and Medical Sciences, Strandboulevarden 49, DK-2100 Copenhagen Ø, Denmark.
Department of Pathology, Copenhagen University Hospital, Blegdamsvej 9, DK-2100 Copenhagen Ø, Denmark.
Sci Rep. 2014 Mar 7;4:4313. doi: 10.1038/srep04313.
Platinum chemotherapy remains part of standard therapies in the management of a variety of cancers. Severe side effects and a high degree of resistance to platinum drugs have led numerous researchers to search for predictive biomarkers, which could aid in identifying patients that are the most likely to respond to therapy. The ERCC1-ERCC4 endonuclease plays a critical role in the repair of platinum-DNA damage and has widely been studied in relation to sensitivity to platinum chemotherapy. The standard method to evaluate ERCC1 protein expression is through the use of immunohistochemistry with monoclonal antibody 8F1, an antibody that was recently found to bind an unrelated protein. The present study determines the specificity of a novel antibody, monoclonal antibody 4F9, and presents a method to evaluate ERCC1 expression in colorectal tumor specimens. Using relevant cell lines as controls, the specificity of antibody 4F9 was tested by immunoblotting, immunohistochemistry and immunofluorescence. Scoring guidelines to aid in the evaluation of ERCC1 tumor expression were developed and evaluated in archival formalin-fixed paraffin embedded colorectal cancer specimens. Antibody 4F9 was found to be specific by all methods applied and it was possible to evaluate the ERCC1 expression in the majority (85%) of colorectal cancer tumor specimens.
铂类化疗仍然是多种癌症治疗中标准疗法的一部分。严重的副作用和对铂类药物的高度耐药性促使众多研究人员寻找预测性生物标志物,这有助于识别最可能对治疗产生反应的患者。ERCC1-ERCC4核酸内切酶在铂-DNA损伤修复中起关键作用,并且已针对铂类化疗敏感性进行了广泛研究。评估ERCC1蛋白表达的标准方法是使用单克隆抗体8F1进行免疫组织化学,而该抗体最近被发现可结合一种不相关的蛋白质。本研究确定了一种新型抗体单克隆抗体4F9的特异性,并提出了一种评估结直肠肿瘤标本中ERCC1表达的方法。以相关细胞系作为对照,通过免疫印迹、免疫组织化学和免疫荧光检测抗体4F9的特异性。制定了有助于评估ERCC1肿瘤表达的评分指南,并在存档的福尔马林固定石蜡包埋的结直肠癌标本中进行了评估。通过所有应用方法发现抗体4F9具有特异性,并且有可能在大多数(85%)结直肠癌肿瘤标本中评估ERCC1表达。