Effect of Chemotherapeutic Drugs on Caspase-3 Activity, as a Key Biomarker for Apoptosis in Ovarian Tumor Cell Cultured as Monolayer. A Pilot Study

Abstract

We aimed to develop a cost-effective and robust method to predict drug resistance in individualpatients. Representative tissue fragments were obtained from tumors removed from femalepatients, aged 24-74 years old. The tumor tissue was taken by a histopathology's or a surgeonunder sterile conditions. Cells obtained by enzymatic dissociation from tumor after surgery, werecultured as a monolayer for 6 days. Paclitaxel, doxorubicin, carboplatin and endoxan alone orin combination were added at the beginning of culture and after 6 days, Alamar blue test wasused for showing action on cell proliferation why caspase- 3 activity assays for verifying actionon apoptosis. Inhibitory action on cell proliferation was noted in 2 of 12 patients tumor treatedwith both single and combined drugs. Using caspase-3 assay we showed that 50% of tumor cellswas resistant to single chemotherapeutic drugs and 40% for combined. In 2 of 12 tumors, whichdid not reacted on single drugs, positive synergistic action on cell proliferation was observedin combination of D + E and C + E. This pilot study suggests: 1) monolayer culture of tumorcells, derived from individual patients, before chemotherapy could provide a suitable model forstudying resistance for drugs; 2) caspase-3 activity is cheap and useful methods; 3) Alamar bluetest should be taken into consideration for measuring cell proliferation.