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Antiproliferative and apoptotic effect of ethanolic extract of Calocybe indica on PANC-1 and MIAPaCa2 cell lines of pancreatic cancer.

BACKGROUND
Pancreatic cancer is an aggressive malignancy with poor prognosis. New options for its treatment including the use of the substances of natural origin are now on the agenda.


AIM
An attempt has been taken to examine the effect of ethanolic extract (EE) of edible mushroom Calocybe indica on PANC-1 and MIAPaCa2 cell lines of pancreatic cancer in vitro.


MATERIALS AND METHODS
Study of cell morphology, evaluation of apoptosis by DAPI staining under florescence microscope, anti-proliferative study by MTT assay, lactate dehydrogenase assay, pro-apoptotic and anti-apoptotic protein detection by Western blotting were performed to test effects of Calocybe indica EE on the cells of these two lines. Effect on in vitro migration was studied by scratch method.


RESULTS
Both cell lines treated with 250 or 500 µg/mL of EE underwent morphological changes - became more round and shrunken, with membrane blebbing and the reduced cell confluence. The nuclei of treated cells became condensed and fragmented. The percentage of apoptotic cells increased as EE concentration increased from 100 µg/mL to 500 µg/mL. Anti-proliferative effect was recorded at all concentrations within 100-750 µg/mL. The effect EE on lactate dehydrogenase leakage was concentration-dependent in both cell lines. Western blotting showed that caspase-3 and -9, and p53 protein levels were increased and BcL2 protein was decreased after treatment of the cells with EE compared to the control. PANC-1 cell migration was inhibited by 80.12 ± 4.25% after 24 h treatment of the cells with 500 µg/mL EE compared to the control.


CONCLUSIONS
EE of Calocybe indica inhibits the growth and induces apoptosis in pancreatic cancer cell lines in vitro.