Wang Zhiwei, Song Wen, Aboukameel Amro, Mohammad Mussop, Wang Guoping, Banerjee Sanjeev, Kong Dejuan, Wang Shaomeng, Sarkar Fazlul H, Mohammad Ramzi M
Department of Pathology, Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA.
Int J Cancer. 2008 Aug 15;123(4):958-66. doi: 10.1002/ijc.23610.
Bcl-2 family of proteins plays critical roles in human cancers, including pancreatic cancer, suggesting that the discovery of specific agents targeting Bcl-2 family proteins would be extremely valuable for pancreatic cancer therapy. We have previously reported the synthesis and characterization of TW-37, which seems to be a negative regulator of Bcl-2. In this investigation, we tested our hypothesis whether TW-37 could be an effective inhibitor of cell growth, invasion and angiogenesis in pancreatic cancer cells. Using multiple cellular and molecular approaches such as MTT assay, apoptosis enzyme-linked immunosorbent assay, real-time reverse transcription-polymerase chain reaction, Western blotting, electrophoretic mobility shift assay for measuring DNA binding activity of NF-kappaB, migration, invasion and angiogenesis assays, we found that TW-37, in nanomolar concentrations, inhibited cell growth in a dose- and time-dependent manner. This was accompanied by increased apoptosis and concomitant attenuation of NF-kappaB, and downregulation of NF-kappaB downstream genes such as MMP-9 and VEGF, resulting in the inhibition of pancreatic cancer cell migration, invasion and angiogenesis in vitro and caused antitumor activity in vivo. From these results, we conclude that TW-37 is a potent inhibitor of progression of pancreatic cancer cells, which could be due to attenuation of Bcl-2 cellular signaling processes. Our findings provide evidence showing that TW-37 could act as a small-molecule Bcl-2 inhibitor on well-characterized pancreatic cancer cells in culture as well as when grown as tumor in a xenograft model. We also suggest that TW-37 could be further developed as a potential therapeutic agent for the treatment of pancreatic cancer.
Bcl-2蛋白家族在包括胰腺癌在内的人类癌症中发挥着关键作用，这表明发现靶向Bcl-2蛋白家族的特定药物对胰腺癌治疗具有极高价值。我们之前报道了TW-37的合成与特性，它似乎是Bcl-2的负调节因子。在本研究中，我们测试了TW-37是否可能是胰腺癌细胞生长、侵袭和血管生成的有效抑制剂这一假设。通过多种细胞和分子方法，如MTT法、凋亡酶联免疫吸附测定、实时逆转录-聚合酶链反应、蛋白质免疫印迹法、用于测量NF-κB DNA结合活性的电泳迁移率变动分析、迁移、侵袭和血管生成测定，我们发现纳摩尔浓度的TW-37以剂量和时间依赖性方式抑制细胞生长。这伴随着凋亡增加以及NF-κB的相应减弱，以及NF-κB下游基因如MMP-9和VEGF的下调，导致体外抑制胰腺癌细胞迁移、侵袭和血管生成，并在体内产生抗肿瘤活性。从这些结果我们得出结论，TW-37是胰腺癌细胞进展的有效抑制剂，这可能是由于Bcl-2细胞信号传导过程的减弱。我们的研究结果提供了证据，表明TW-37在培养的特征明确的胰腺癌细胞中以及在异种移植模型中作为肿瘤生长时可作为小分子Bcl-2抑制剂发挥作用。我们还建议TW-37可进一步开发成为治疗胰腺癌的潜在治疗药物。
