Strizzi Luigi, Sandomenico Annamaria, Margaryan Naira V, Focà Annalia, Sanguigno Luca, Bodenstine Thomas M, Chandler Grace S, Reed David W, Gilgur Alina, Seftor Elisabeth A, Seftor Richard E B, Khalkhali-Ellis Zhila, Leonardi Antonio, Ruvo Menotti, Hendrix Mary J C
Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
Oncotarget. 2015 Oct 27;6(33):34071-86. doi: 10.18632/oncotarget.6049.
Nodal is highly expressed in various human malignancies, thus supporting the rationale for exploring Nodal as a therapeutic target. Here, we describe the effects of a novel monoclonal antibody (mAb), 3D1, raised against human Nodal. In vitro treatment of C8161 human melanoma cells with 3D1 mAb shows reductions in anchorage-independent growth and vasculogenic network formation. 3D1 treated cells also show decreases of Nodal and downstream signaling molecules, P-Smad2 and P-ERK and of P-H3 and CyclinB1, with an increase in p27. Similar effects were previously reported in human breast cancer cells where Nodal expression was generally down-regulated; following 3D1 mAb treatment, both Nodal and P-H3 levels are reduced. Noteworthy is the reduced growth of human melanoma xenografts in Nude mice treated with 3D1 mAb, where immunostaining of representative tumor sections show diminished P-Smad2 expression. Similar effects both in vitro and in vivo were observed in 3D1 treated A375SM melanoma cells harboring the active BRAF(V600E) mutation compared to treatments with IgG control or a BRAF inhibitor, dabrafenib. Finally, we describe a 3D1-based ELISA for the detection of Nodal in serum samples from cancer patients. These data suggest the potential of 3D1 mAb for selecting and targeting Nodal expressing cancers.
Nodal在多种人类恶性肿瘤中高表达，因此支持了将Nodal作为治疗靶点进行探索的理论依据。在此，我们描述了一种针对人Nodal产生的新型单克隆抗体（mAb）3D1的作用。用3D1 mAb体外处理C8161人黑色素瘤细胞，结果显示其非锚定依赖性生长和血管生成网络形成减少。经3D1处理的细胞还显示Nodal及下游信号分子P-Smad2和P-ERK以及P-H3和细胞周期蛋白B1减少，而p27增加。此前在人乳腺癌细胞中也报道过类似效应，在这些细胞中Nodal表达通常下调；用3D1 mAb处理后，Nodal和P-H3水平均降低。值得注意的是，用3D1 mAb处理的裸鼠体内人黑色素瘤异种移植物生长减缓，代表性肿瘤切片的免疫染色显示P-Smad2表达减少。与用IgG对照或BRAF抑制剂达拉非尼处理相比，在携带活性BRAF(V600E)突变的3D1处理的A375SM黑色素瘤细胞中，在体外和体内均观察到类似效应。最后，我们描述了一种基于3D1的ELISA方法，用于检测癌症患者血清样本中的Nodal。这些数据表明3D1 mAb在筛选和靶向表达Nodal肿瘤方面具有潜力。
