Antiproliferative, Apoptotic, and Antimigration Property of Ethyl Acetate Extract of Calocybe indica against HeLa and CaSki Cell Lines of Cervical Cancer, and its Antioxidant and Mycochemistry Analysis

Abstract

Background: The incidence rate of cervical cancer is increasing and its existing drugs are becoming more and more resistant. Therefore, we extracted the fruiting body of Calocybe indica edible mushroom in 90% ethyl acetate extract (EAE) and evaluated it as an anticancer property against HeLa and CaSki. Method: We performed cytotoxicity assay by MTT, cell morphological study by phase contrast microscope, and apoptosis study by nuclear morphology via DAPI staining under inverted microscopy; the expressions of proapoptotic and antiapoptotic genes and p53 were examined by Western blotting, cell cycle analysis, and cologenic and cell migration assay. Antioxidant content and activity assays were performed and for mycochemistry analysis of EAE, thin layer chromatography (TLC) was done. Results: EAE-treated HeLa and CaSki cells became round and showed condensed and fragmented nuclei. They inhibited the cell proliferation of both cancer cell lines in a dose-dependent manner. At maximum dose (1250 μg/mL) after 24 h, the cell inhibition percentages of HeLa and CaSki cells were 97.12±10.01 and 98.52±10.08 (p <0.05), respectively. They upregulated the expression of p53, caspase 3, and caspase 9 while down-regulating BcL2 gene. Cell cycle became arrested at G2/M checkpoint of both cancer cell lines by EAE. EAE inhibited colony formation and cell migration. The antioxidant assay showed that EAE contained good amounts of phenolic compounds, flavonoids, and ascorbic acids and had good antioxidant activity. TLC supported the presence of bioactive components. Conclusion: The EAE of C. indica exerts very potent anticervical cancer effects. It is urgent that future studies analyze its bioactive compounds in detail and examine them in animal models.