Ethyl Acetate Extract of Colletotrichum gloeosporioides Promotes Cytotoxicity and Apoptosis in Human Breast Cancer Cells

Abstract

Fungal endophytes are known to be a paragon for producing bioactive compounds with a variety of pharmacological importance. The current study aims to elucidate the molecular alterations induced by the bioactive compounds produced by the fungal endophyte Colletotrichum gloeosporioides in the tumor microenvironment of human breast cancer cells. GC/MS analysis of the ethyl acetate (EA) extract of C. gloeosporioides revealed the presence of bioactive compounds with anticancer activity. The EA extract of C. gloeosporioides exerted potential plasmid DNA protective activity against hydroxyl radicals of Fenton’s reagent. The cytotoxic activity further revealed that MDA-MB-231 cells exhibit more sensitivity toward the EA extract of C. gloeosporioides as compared to MCF-7 cells, whereas non-toxic to non-cancerous HEK293T cells. Furthermore, the anticancer activity demonstrated by the EA extract of C. gloeosporioides was studied by assessing nuclear morphometric analysis and induction of apoptosis in MDA-MB-231 and MCF-7 cells. The EA extract of C. gloeosporioides causes the alteration in cellular and nuclear morphologies, chromatin condensation, long-term colony inhibition, and inhibition of cell migration and proliferation ability of MDA-MB-231 and MCF-7 cells. The study also revealed that the EA extract of C. gloeosporioides treated cells undergoes apoptosis by increased production of reactive oxygen species and significant deficit in mitochondrial membrane potential. Our study also showed that the EA extract of C. gloeosporioides causes upregulation of pro-apoptotic (BAX, PARP, CASPASE-8, and FADD), cell cycle arrest (P21), and tumor suppressor (P53) related genes. Additionally, the downregulation of antiapoptotic genes (BCL-2 and SURVIVIN) and increased Caspase-3 activity suggest the induction of apoptosis in the EA extract of C. gloeosporioides treated MDA-MB-231 and MCF-7 cells. Overall, our findings suggest that the bioactive compounds present in the EA extract of C. gloeosporioides promotes apoptosis by altering the genes related to the extrinsic as well as the intrinsic pathway. Further in vivo study in breast cancer models is required to validate the in vitro observations.

Isolation and characterization of N‐(2‐Hydroxyethyl)hexadecanamide from Colletotrichum gloeosporioides with apoptosis‐inducing potential in breast cancer cells

“…Fungal endophytes are an exceptional source of biologically active compounds that can be explored as emerging sources of pharmacological lead compounds in the development of new therapeutics for cancer. Considering the potential antioxidant and anticancer potential of EA extract of C. gloeosporioides, 24,25 we aimed to purify the monomeric compound responsible for triggering apoptosis in MDA-MB-231 and MCF-7 cells. In light of this, bioactive fraction 1 was identified as PEA using various spectroscopy techniques followed by antioxidant activityguided fractionation of EA extract of C. gloeosporioides.…”

“…Nuclear morphometric analysis using a fluorescent dye DAPI was used to evaluate the presence of apoptotic cells and nuclear morphology of breast cancer cells, MDA-MB-231 and MCF-7 treated with PEA. 25 Briefly, 1 Â 10 5 cells were seeded in 6-well plate containing sterile cover slip and treated with PEA (IC 30 , IC 50 , and IC 70 ). After 24 h the cells were fixed with paraformaldehyde at room temperature followed by incubation with 70% methanol for 30 min at À20 C. Post-incubation cells were stained with DAPI (Puregene, Genetix) for 10 min at room temperature and the image was captured using fluorescence microscope (Leica, Germany).…”

“…Our previous findings revealed that the bioactive components present in the ethyl acetate (EA) extract of C. gloeosporioides potentiates cytotoxicity against various cancer cells including two human breast cancer cells, MDA-MB-231 and MCF-7. 24,25 Notably, the EA extract of C. gloeosporioides also exacerbates apoptosis in human breast cancer cells by modulating the expression level of apoptosis-related genes and inducing the activity of Caspase-3. Therefore, considering our previous findings, 24,25 we were interested to isolate and characterize the bioactive compounds produced from C. gloeosporioides responsible for anti-breast cancer efficiency, which has not yet been reported.…”

“…24,25 Notably, the EA extract of C. gloeosporioides also exacerbates apoptosis in human breast cancer cells by modulating the expression level of apoptosis-related genes and inducing the activity of Caspase-3. Therefore, considering our previous findings, 24,25 we were interested to isolate and characterize the bioactive compounds produced from C. gloeosporioides responsible for anti-breast cancer efficiency, which has not yet been reported. In the present study, we performed antioxidant activity-guided fractionation of the EA extract of C. gloeosporioides, as the antioxidant properties may be responsible for the anticancer activity.…”

Exploration of in vitro cytotoxic and in ovo antiangiogenic activity of ethyl acetate extract of Penicillium oxalicum

“…54 According to several studies, the MDA-MB-231 and MCF-7 cells induced apoptosis and displayed the typical morphological changes associated with apoptosis, including cell shrinkage, nuclei blebbing, chromatin condensation, and nuclear fragmentation. [55][56][57][58] Furthermore, the increased expression of genes associated with apoptosis and the stimulation of the pro-apoptotic protein Caspase-3 evidence that the EA extract of P. oxalicum facilitates apoptosis in breast cancer cells. This suggests that the bioactive compound present in the EA extract of P. oxalicum has potential to induce apoptosis, a crucial mechanism for suppressing the growth of breast cancer.…”