Treatment with gemcitabine and TRA-8 anti-death receptor-5 mAb reduces pancreatic adenocarcinoma cell viability in vitro and growth in vivo
Treatment with gemcitabine and TRA-8 anti-death receptor-5 mAb reduces pancreatic adenocarcinoma cell viability in vitro and growth in vivo
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DOI：
10.1016/j.gassur.2006.08.007
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发表时间：
2006-11-01
影响因子：
3.2
通讯作者：
Vickers, Selwyn M.
中科院分区：
文献类型：
--
作者：
DeRosier, Leo Christopher;Huang, Zhi-Qiang;Vickers, Selwyn M.
吉西他滨是治疗胰腺癌的一线药物，但生存率很低。本研究利用两种人胰腺癌细胞系S2VP10和MIA PaCa-2，评估了抗人死亡受体5单克隆抗体（TRA-8）联合吉西他滨在体外和体内的作用。采用胰腺癌皮下模型进行体内疗效试验。用不同剂量的吉西他滨和TRA-8处理S2VP10和MIA PaCa-2细胞。分别用三磷酸腺苷法和膜联蛋白V染色法测定细胞活力和凋亡。对JC-1染色细胞进行荧光活化细胞分选分析，评估线粒体膜的不稳定性。Western blot检测Caspase活性。评估MIA PaCa-2裸鼠皮下异种移植物对200 μ g TRA-8（在植入后第9、13、16、20、23和27天腹腔注射）和120 mg/kg吉西他滨（在第10、17和24天ig）治疗的反应。用卡尺测量肿瘤生长情况。与单独使用任何一种药物相比，接受TRA-8和吉西他滨联合治疗的MIA PaCa-2和S2VP10细胞表现出增强的细胞毒性、膜联蛋白V染色和线粒体不稳定。与单独使用任何一种药物相比，联合治疗在两种细胞系中都产生了增强的caspase-3和-8激活。体内研究表明，未治疗38天，吉西他滨治疗32天，TRA-8治疗49天，联合治疗64天，平均皮下肿瘤表面积（产生两个最大直径）翻倍。TRA-8是一种诱导凋亡的激动性单克隆抗体，在体外通过增强半胱天冬酶激活与吉西他滨联合产生协同细胞毒性。这些发现，在接受联合治疗的小鼠胰腺癌异种移植模型中，肿瘤生长受到了实质性的抑制，这对于抗死亡受体治疗胰腺癌是令人鼓舞的。
Gemcitabine is a first line agent for pancreatic cancer, but yields minimal Survival benefit. This study evaluated in vitro and in vivo effects of a monoclonal antibody (TRA-8) to human death receptor 5, combined with gemcitabine, using two human pancreatic cancer cell lines, S2VP10 and MIA PaCa-2. A subcutaneous model of pancreatic cancer was employed to test in vivo efficacy. S2VP10 and MIA PaCa-2 cells were treated with varying doses of gemcitabine and TRA-8. Cell viability and apoptosis were determined with an adenosine triphosphate assay and annexin V staining, respectively. Mitochondrial membrane destabilization was evaluated with fluorescence-activated cell sorting analysis of JC-1 stained cells. Caspase activation was evaluated by Western blot analysis. MIA PaCa-2 subcutaneous xenografts in athymic nude mice were evaluated for response to treatment with 200 mu g of TRA-8 (intraperitoneal on days 9, 13, 16, 20, 23, and 27 postimplant) and 120 mg/kg gemcitabine (I.P. on days 10, 17, and 24). Tumor growth was measured with calipers. MIA PaCa-2 and S2VP10 cells receiving combination treatment with TRA-8 and gemcitabine demonstrated enhanced cytotoxicity, annexin V staining, and mitochondrial destabilization compared to either agent alone. Combination treatment produced enhanced caspase-3 and -8 activation in both cell lines compared with either agent alone. In vivo studies demonstrated mean subcutaneous tumor surface area (produce of two largest diameters) doubling times of 38 days untreated, 32 days gemcitabine, 49 days TRA-8, and 64 days combination treatment. TRA-8 is In apoptosis-inducing agonistic monoclonal antibody that produced synergistic cytotoxicity in combination with gemcitabine in vitro through enhanced caspase activation. These findings, with substantial inhibition of tumor growth in a mouse pancreatic cancer xenograft model receiving combination therapy, are encouraging for anti-death receptor therapy in the treatment of pancreatic cancer.