School of Medical Science and Technology, Indian Institute of Technology, Kharagpur, India.
Life Sci. 2012 Oct 22;91(15-16):789-99. doi: 10.1016/j.lfs.2012.08.024. Epub 2012 Aug 24.
Combinatorial therapies that target multiple signaling pathways may provide improved therapeutic responses over monotherapies. In the present study, we evaluated the effect of celecoxib and AEE788 alone and in combination on cell proliferation, invasion, migration, angiogenesis, morphological changes, actin filament organization and apoptosis induction in the human colon cancer cell lines.
Effect of celecoxib and AEE788 alone and in combination on colon cancer cell lines was evaluated by cell proliferation assay, morphological analysis, cell cycle analysis, scratch-wound healing and chorioallantoic membrane assays, zymography, nuclear fragmentation and western blot analyses.
Either drug alone or in combination inhibited human colonic adenocarcinoma cell lines HCT 15 and HT 29 in a dose-dependent manner. Microscopic analysis revealed inhibition of cell membrane extensions, cell shrinkage, and disorganization of actin filaments. Additionally, either drug alone or in combination inhibited HCT 15 migration, invasion and angiogenesis by suppressing matrix metalloproteinase-2 and -9 activities. Increased reactive oxygen generation, loss of mitochondrial membrane potential, cleavage of PARP, caspase-3 activation and DNA ladder formation characterized the induction of apoptosis by celecoxib and/or AEE788 treatment. Either drug individually induced apoptosis via down-regulation of the anti-apoptotic proteins Bcl(2) and Bcl-x(L), and up-regulation of pro-apoptotic protein Bax, cleavage of PARP, activation of caspase-3 and inhibition of vascular endothelial growth factor receptor signaling pathways.
Results indicate that AEE788 potentiates celecoxib-mediated inhibition of proliferation and angiogenesis in HCT 15 colon cancer cells and may prove useful for developing a combinatorial therapy for colon cancer.
靶向多个信号通路的联合疗法可能比单药治疗提供更好的治疗反应。在本研究中，我们评估了塞来昔布和 AEE788 单独和联合应用对人结肠癌细胞系增殖、侵袭、迁移、血管生成、形态变化、肌动蛋白丝组织和细胞凋亡诱导的影响。
通过细胞增殖测定、形态分析、细胞周期分析、划痕愈合和绒毛尿囊膜测定、酶谱分析、核片段化和 Western blot 分析评估塞来昔布和 AEE788 单独和联合应用对结肠癌细胞系的影响。
两种药物单独或联合使用均可剂量依赖性地抑制人结肠腺癌细胞系 HCT 15 和 HT 29。显微镜分析显示，细胞膜延伸、细胞收缩和肌动蛋白丝的紊乱受到抑制。此外，两种药物单独或联合使用均可抑制 HCT 15 迁移、侵袭和血管生成，抑制基质金属蛋白酶-2 和 -9 的活性。塞来昔布和/或 AEE788 处理诱导细胞凋亡的特征是活性氧生成增加、线粒体膜电位丧失、PARP 裂解、caspase-3 激活和 DNA 梯形成。两种药物单独通过下调抗凋亡蛋白 Bcl(2)和 Bcl-x(L)，上调促凋亡蛋白 Bax，PARP 裂解，caspase-3 激活和抑制血管内皮生长因子受体信号通路，诱导细胞凋亡。
结果表明，AEE788 增强了塞来昔布对 HCT 15 结肠癌细胞增殖和血管生成的抑制作用，可能有助于开发结肠癌的联合治疗方法。
