Pilot Study Of The Mechanism Of Antitumor Agent Carboxyamido-triazole's Cell Proliferation Inhibiting Effect

Carboxyamido-triazole(CAI) is an experimental anti-cancer agent being researched and developed by our laboratory which had entered clinical trial stage. Up to now, many articles have been published about the mechanism sutdy of this agent. The published results showed that CAI can inhibit the proliferation of endothelial cell and many tumor cell lines. CAI can inhibit calcium influx stimulated by bFGF, VEGF, TG and other agents in endothelial cell and some tumor cell lines. CAI can inhibit angiogenesis in the chick chorioallantoic membrane assay and rat aortic ring assay. CAI can inhibit the growth and invasion of many tumor xenograft animal models.Although much research work has been done, little is known about the molecular and cellular mechanism of CAI's cell proliferation inhibiting effect. Therefore, the aim of our study is to clarify the possible mechanism of CAI's cell proliferation inhibiting effect at molecular and cellular level. Our study mainly focuses on two aspects below:1. The influence of CAI on cell proliferation, apoptosis and cell cycle regulation;2. The influence of CAI on cAMP signal transduction pathway and the influence of intracellular cAMP elevating agents(forskolin, theophylline and IBMX) on CAI's cell proliferation inhibiting effect;The methods adopted in this study:1. This study adopted cell growth curve assay, WST-8 assay and ~3H-thymidine incorporation assay to evaluate the effect of CAI on SH-SY5Y cell proliferation, viability and DNA synthesis; this study adopted PI staining and flow cytometry to evaluate the effect of CAI on SH-SY5Y cell apoptosis and cell cycle regulation; this study adopted trypan blue exclusion assay to evaluate the induction effect of...