The effects of intracellular calcium increase on apoptosis based on intracellular mechanisms

In this thesis, apoptotic effects of intracellular calcium increase were investigated by genetaring endoplasmic reticulum stress in SKOV-3 ovarian cancer cells while integrins were activated. During the SKOV-3 cells were adhering to 50 g/ml fibronectin, endoplasmic reticulum stress was formed with adding 24 M tunicamycin and incubated 30 or 60 minutes. Afterwards, intracellular Ca+2 increase and mobility were imaged by marking with Fluo-3 AM and Rhod-2 AM dyes in fluorescent microscope. In addition to, the binding capacity of SKOV-3 cells to fibronectin was investigated by using Real Time Cell Analyser (RTCA) system and the results obtained here were showed that the adhesion of cells decreased dependend on time and dose of tunicamycin. .In SKOV-3 cells whose integrin activation level increased, mRNA expression levels of Bax, BcL-2, Bcl-XL, cytocrome c, EndoG, AIF, calpain 1,calpain 2, caspase 3, caspase 8 and FAK genes were also investigated by RT-PCR. These result indicated that in our system, SKOV-3 cells might be using the various apoptotic pathways rather than one. In conclusion, forming stress in endoplasmic reticulum induced apoptosis in high metastatic SKOV-3 cells. The results obtained from this study would be useful for drug development intended for therapy of these cancer type whose mortalite rate is high.