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

Document Type: Research article

Authors

1 Deartament fo Physiology and Toxicology of Reproduction, Institute of Zoology, Jagiellonian University in Krakow

2 1Department of Physiology and Toxicology of Reproduction, Institute of Zoology, Jagiellonian University, 30-389 Krakow, Gronostajowa 9, Poland

3 2Department of Tumor Pathology and 3Department of Gynecological Oncology, Center of Oncology, Maria Sklodowska-Curie Memorial Institute, Krakow, Garncarska 11, Poland

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.

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