title: 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 individual patients. Representative tissue fragments were obtained from tumors removed from female patients, aged 24-74 years old. The tumor tissue was taken by a histopathology's or a surgeon under sterile conditions. Cells obtained by enzymatic dissociation from tumor after surgery, were cultured as a monolayer for 6 days. Paclitaxel, doxorubicin, carboplatin and endoxan alone or in combination were added at the beginning of culture and after 6 days, Alamar blue test was used for showing action on cell proliferation why caspase- 3 activity assays for verifying action on apoptosis. Inhibitory action on cell proliferation was noted in 2 of 12 patients tumor treated with both single and combined drugs. Using caspase-3 assay we showed that 50% of tumor cells was resistant to single chemotherapeutic drugs and 40% for combined. In 2 of 12 tumors, which did not reacted on single drugs, positive synergistic action on cell proliferation was observed in combination of D + E and C + E. This pilot study suggests: 1) monolayer culture of tumor cells, derived from individual patients, before chemotherapy could provide a suitable model for studying resistance for drugs; 2) caspase-3 activity is cheap and useful methods; 3) Alamar blue test should be taken into consideration for measuring cell proliferation.
authors.affiliation: Department of Physiology and Toxicology of Reproduction, Institute of Zoology, Jagiellonian University, 30-389 Krakow, Gronostajowa 9, Poland.
authors.forename: Ewa L
authors.initials: EL
authors.lastname: Gregoraszczuk
authors.affiliation: Department of Physiology and Toxicology of Reproduction, Institute of Zoology, Jagiellonian University, 30-389 Krakow, Gronostajowa 9, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Rak-Mardyła
authors.affiliation: Department of Tumor Pathology.
authors.forename: Janusz
authors.initials: J
authors.lastname: Ryś
authors.affiliation: Department of Gynecological Oncology, Center of Oncology, Maria Sklodowska-Curie Memorial Institute, Krakow, Garncarska 11, Poland.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Jakubowicz
authors.affiliation: Department of Gynecological Oncology, Center of Oncology, Maria Sklodowska-Curie Memorial Institute, Krakow, Garncarska 11, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Urbański
keywords: Alamar Blue test; Casapase-3 activity; Chemotherapeutic drugs; Monolayer cell culture; Ovarian cancer