Nicoletti Roberta, Lopez Salvatore, Bellone Stefania, Cocco Emiliano, Schwab Carlton L, Black Jonathan D, Centritto Floriana, Zhu Liancheng, Bonazzoli Elena, Buza Natalia, Hui Pei, Mezzanzanica Delia, Canevari Silvana, Schwartz Peter E, Rutherford Thomas J, Santin Alessandro D
Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, 06520, USA.
Clin Exp Metastasis. 2015 Jan;32(1):29-38. doi: 10.1007/s10585-014-9688-8. Epub 2014 Nov 15.
Ovarian and uterine carcinosarcoma (CS) are characterized by their aggressive clinical behavior and poor prognosis. We evaluated the efficacy of trastuzumab-emtansine (T-DM1), against primary HER2 positive and HER2 negative CS cell lines in vitro and in vivo. Eight primary CS cell lines were evaluated for HER2 amplification and protein expression by fluorescence in situ hybridization, immunohistochemistry, flow cytometry and qRT-PCR. Sensitivity to T-DM1-induced antibody-dependent-cell-mediated-cytotoxicity (ADCC) was evaluated in 4-h-chromium-release-assays. T-DM1 cytostatic and apoptotic activities were evaluated using flow cytometry based proliferation assays. In vivo activity of T-DM1 was also evaluated. HER2 protein overexpression and gene amplification were detected in 25 % (2/8) of the primary CS cell lines. T-DM1 and T were similarly effective in inducing strong ADCC against CS overexpressing HER2 at 3+ levels. In contrast, T-DM1 was dramatically more effective than T in inhibiting cell proliferation (P < 0.0001) and in inducing G2/M phase cell cycle arrest in the HER2 expressing cell lines (shift of G2/M: mean ± SEM from 14.87 ± 1.23 to 66.57 ± 4.56 %, P < 0.0001). Importantly, T-DM1 was highly active at reducing tumor formation in vivo in CS xenografts overexpressing HER2 (P = 0.0001 and P < 0.0001 compared to T and vehicle respectively) with a significantly longer survival when compared to T and vehicle mice (P = 0.008 and P = 0.0001 respectively). T-DM1 may represent a novel treatment option for the subset of HER2 positive CS patients with disease refractory to chemotherapy.
卵巢和子宫癌肉瘤（CS）具有侵袭性的临床行为和较差的预后。我们评估了曲妥珠单抗-恩美曲妥珠单抗（T-DM1）在体外和体内对原发性HER2阳性和HER2阴性CS细胞系的疗效。通过荧光原位杂交、免疫组织化学、流式细胞术和qRT-PCR评估了8种原发性CS细胞系的HER2扩增和蛋白表达。在4小时铬释放试验中评估了对T-DM1诱导的抗体依赖性细胞介导的细胞毒性（ADCC）的敏感性。使用基于流式细胞术的增殖试验评估了T-DM1的细胞生长抑制和凋亡活性。还评估了T-DM1的体内活性。在25%（2/8）的原发性CS细胞系中检测到HER2蛋白过表达和基因扩增。T-DM1和曲妥珠单抗（T）在诱导针对HER2过表达水平为3+的CS的强烈ADCC方面同样有效。相比之下，T-DM1在抑制细胞增殖（P<0.0001）和诱导HER2表达细胞系中的G2/M期细胞周期停滞方面比T显著更有效（G2/M期的变化：平均值±SEM从14.87±1.23变为66.57±4.56%，P<0.0001）。重要的是，T-DM1在体内对过表达HER2的CS异种移植瘤的肿瘤形成具有高度活性（与T和溶剂相比，P分别为0.0001和P<0.0001），与T和溶剂处理的小鼠相比，生存期显著延长（分别为P=0.008和P=0.0001）。T-DM1可能是对化疗难治性疾病的HER2阳性CS患者亚组的一种新的治疗选择。
