Nakajima Yasuhiro, Kanno Takeshi, Nagaya Toshio, Kuribayashi Kozo, Nakano Takashi, Gotoh Akinobu, Nishizaki Tomoyuki
Division of Respiratory Medicine, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan.
Cell Physiol Biochem. 2015;35(1):51-60. doi: 10.1159/000369674. Epub 2015 Jan 2.
BACKGROUND/AIMS: Malignant pleural mesothelioma (MPM) is an aggressive malignant tumor and an effective therapy has been little provided as yet. The present study investigated the possibility for the adenosine deaminase (ADA) inhibitor EHNA as a target of MPM treatment.
MTT assay, TUNEL staining, monitoring of intracellular adenosine concentrations, and Western blotting were carried out in cultured human MPM cell lines without and with knocking-down ADA. The in vivo effect of EHNA was assessed in mice inoculated with NCI-H2052 MPM cells.
EHNA induced apoptosis of human MPM cell lines in a concentration (0.01-1 mM)- and treatment time (24-48 h)-dependent manner, but such effect was not obtained with another ADA inhibitor pentostatin. EHNA increased intracellular adenosine concentrations in a treatment time (3-9 h)-dependent manner. EHNA-induced apoptosis of MPM cells was mimicked by knocking-down ADA, and the effect was neutralized by the adenosine kinase inhibitor ABT-702. EHNA clearly suppressed tumor growth in mice inoculated with NCI-H2052 MPM cells.
The results of the present study show that EHNA induces apoptosis of MPM cells by increasing intracellular adenosine concentrations, to convert to AMP, and effectively prevents MPM cell proliferation. This suggests that EHNA may be useful for treatment of the tragic neoplasm MPM.
背景/目的:恶性胸膜间皮瘤(MPM)是一种侵袭性恶性肿瘤,目前尚未有有效的治疗方法。本研究探讨了腺苷脱氨酶(ADA)抑制剂EHNA作为MPM治疗靶点的可能性。
在未敲低和敲低ADA的培养人MPM细胞系中进行MTT试验、TUNEL染色、细胞内腺苷浓度监测和蛋白质印迹分析。在接种NCI-H2052 MPM细胞的小鼠中评估EHNA的体内作用。
EHNA以浓度(0.01-1 mM)和处理时间(24-48小时)依赖性方式诱导人MPM细胞系凋亡,但另一种ADA抑制剂喷司他丁未获得此效果。EHNA以处理时间(3-9小时)依赖性方式增加细胞内腺苷浓度。敲低ADA可模拟EHNA诱导的MPM细胞凋亡,腺苷激酶抑制剂ABT-702可中和该作用。EHNA明显抑制接种NCI-H2052 MPM细胞的小鼠的肿瘤生长。
本研究结果表明,EHNA通过增加细胞内腺苷浓度诱导MPM细胞凋亡,使其转化为AMP,并有效阻止MPM细胞增殖。这表明EHNA可能对治疗这种恶性肿瘤MPM有用。