Department of Hematology, Tongji Hospital, Tongji University, Shanghai 200065, China.
Acta Pharmacol Sin. 2012 Dec;33(12):1557-62. doi: 10.1038/aps.2012.91. Epub 2012 Nov 5.
To study the antitumor effect of anti-NprPSA monoclonal antibody (mAb) in combination with ManNPr, a precursor of N-propionyl PSA, in multiple myeloma (MM), and to explore the mechanisms of the action.
Human multiple myeloma cell line RPMI-8226 was tested. The cells were pre-treated with ManNPr (1, 2, and 4 mg/mL), and then incubated with anti-NprPSA mAb (1 mg/mL). Cell apoptosis in vitro was detected using MTT assay and flow cytometry. BALB/c nude mice were inoculated sc with RPC5.4 cells. On 5 d after the injection, the mice were administered sc with anti-NprPSA mAb (200 μg/d) and ManNPr (5 mg/d) for 8 d. The tumor size and body weight were monitored twice per week. TUNEL assay was used for detecting apoptosis in vivo. The apoptotic pathway involved was examined using Western blot analysis and caspase inhibitor.
Treatment of RPMI-8226 cells with anti-NprPSA mAb alone failed to inhibit cell growth in vitro. In RPMI-8226 cells pretreated with ManNPr, however, the mAb significantly inhibited the cell proliferation, decreased the viability, and induced apoptosis, which was associated with cleavage of caspase-3, caspase-8, caspase-9, and poly(ADP-ribose) polymerase. In the mouse xenograft model, treatment with the mAb in combination with ManNPr significantly inhibited the tumor growth, and induced significant apoptosis as compared to treatment with the mAb alone. Moreover, apoptosis induced by the mAb in vivo resulted from the activation of the caspases and poly(ADP-ribose) polymerase.
The anti-NprPSA mAb in combination with ManNPr is an effective treatment for in vitro and in vivo induction of apoptosis in multiple myeloma.
研究抗 NPRPSA 单克隆抗体（mAb）联合 N-丙酰基 PSA 前体 ManNPr 对多发性骨髓瘤（MM）的抗肿瘤作用，并探讨其作用机制。
检测人多发性骨髓瘤细胞系 RPMI-8226。ManNPr（1、2 和 4 mg/ml）预处理细胞，然后与抗 NPRPSA mAb（1 mg/ml）孵育。MTT 法和流式细胞术检测细胞体外凋亡。BALB/c 裸鼠皮下接种 RPC5.4 细胞。注射后 5 天，皮下注射抗 NPRPSA mAb（200 μg/d）和 ManNPr（5 mg/d），连续 8 天。每周监测肿瘤大小和体重 2 次。TUNEL 法检测体内细胞凋亡。Western blot 分析和 caspase 抑制剂检测凋亡途径。
抗 NPRPSA mAb 单独处理 RPMI-8226 细胞不能抑制细胞体外生长。然而，在 ManNPr 预处理的 RPMI-8226 细胞中，mAb 显著抑制细胞增殖，降低细胞活力，诱导细胞凋亡，与 caspase-3、caspase-8、caspase-9 和多聚（ADP-核糖）聚合酶的裂解有关。在小鼠异种移植模型中，与单独使用 mAb 相比，mAb 联合 ManNPr 治疗显著抑制肿瘤生长，并诱导显著的细胞凋亡。此外，体内 mAb 诱导的细胞凋亡源于 caspase 和多聚（ADP-核糖）聚合酶的激活。
抗 NPRPSA mAb 联合 ManNPr 是体外和体内诱导多发性骨髓瘤细胞凋亡的有效治疗方法。
