Signal Transduction Study In Chemotherapeutic Drug Induced Apoptosis Of Tumor Cell

The major mechanism of chemotherapy is Apoptosis However, the trigger procedure of chemotherapy induced apoptosis cascade is still uncertain More detailed understanding of chemotherapy induced apoptosis is a theoretical foundation for finding out a better choice of clinical protocol In the present study, we aimed to establish an apoptosis experimental system and to achieve further understanding of the molecular mechanism of 5-FU induced apoptosis We analyzed apoptosis procedure of 5-FU treated AGS cell line by cell culture, DNA ladder, flow Cytometry technique Human Cyt-c Apaf-1~ Caspase-3 Bax Bcl-2 gene expression were measured by Northern blot analysis and RT-PCR method In order to evaluate the role of Cyt-c pathway, we established Apaf-1 over expression system by transfecting Apaf-1 cDNA into cultured AGS cell line Our results indicated that the typical apoptosis induced by 5-FU appeared after 24 hours, the appropriate concentration should be o 5minol/L The expression level of major molecular of Cyt-c pathway shows significant discrepancy In this study, we established a set of basic apoptosis induction system and related evaluation methods Our results suggested Cyt-c pathway may play an important role in chemotherapeutic drugs induced apoptosis The early expression of Cyt-c and Apaf-1 may be the key procedure of Cyt-c release from mitochondrial and followed caspases activation...