2-Methoxyestradiol-bis-sulfamate induces apoptosis and autophagy in a tumorigenic breast epithelial cell line

Abstract

In anticancer research where the focus is on finding agents that induces cell death while leaving nontumorigenic cells less affected, a novel 2-methoxyestradiol derivative has come forth. 2-Methoxyestradiol-bissulphamate (2-MeOE2bisMATE) is a 2-methoxyestradiol derivative produced by bis-sulphamoylation which possesses increased antiproliferative activity and biological availability. Several questions remain regarding the the type of cell death mechanisms and possible induction of autophagy by 2-MeOE2bisMATE. The aim of this in vitro study was to investigate the cell death mechanisms exerted by 2-MeOE2bisMATE in an adenocarcinoma cell line (MCF-7) by analyzing its influence on cell growth, morphology and possible induction of cell death. Spectrophotometry (crystal violet staining), transmission electron microscopy (TEM), light microscopy (haematoxylin and eosin staining) and fluorescent microscopy (Hoechst 33342, propidium iodide and acridine orange) were employed. Spectrophotometrical studies indicated that 2-MeOE2bisMATE decreased cell numbers to 75% in MCF-7 cells after 24 h and to 47% after 48 h of exposure. TEM demonstrated membrane blebbing, nuclear fragmentation and chromatin condensation indicating hallmarks of apoptosis. Light microscopy revealed the presence of several cells blocked in metaphase and apoptotic cells were also observed.Fluorescent microscopy demonstrated increased lysosomal staining; suggesting the induction of autophagy. 2-MeOE2bisMATE shows therapeutic potential as an anticancer agent and the investigation of the cell death mechanisms used by 2-MeOE2bisMATE thus warrants further investigation.

Sulphamoylated estradiol analogue induces antiproliferative activity and apoptosis in breast cell lines

“…MCF-7 and MDA-MB-231 cells were cultured in Dulbecco's minimum essential medium eagle (DMEM) and supplemented with 10% heat-inactivated FCS (56ºC, 30 min), 100 U/ml penicillin G, 100 µg/ml streptomycin and 250 µg/l fungizone [9]. MCF-12A maintenance medium consisted of a 1:1 mixture of DMEM and Ham's-F12 medium, 20 ng/ml epidermal growth factor, 100 ng/ml cholera toxin, 10 µg/ml insulin and 500 ng/ml hydrocortisone, supplemented with 10% heat-inactivated FCS (56ºC, 30 min), 100 U/ml penicillin G, 100 µg/ml streptomycin and 250 µg/l fungizone [2].…”

“…It was conducted according to Visagie et al [2,9]. Images were obtained before and after exposure using the Axiovert 40 CFL microscope (Carl Zeiss, Goettingen, Germany).…”

“…2-methoxyestradiol-bis-sulphamate exposure resulted in a G 2 /M block in cell lines, including the estrogen receptor-positive human breast adenocarcinoma cell line MCF-7, the drug-resistant human adenocarcinoma cell line MCF-7 DOX40, and the highly tumorigenic breast carcinoma cell line MDA-MB-231 [9]. Another analogue, 2-methoxyestra-1,3,5(10)16-tetraene-3-carboxamide (also known as ENMD-1198, C24, 883, or ENMD-0998) exerts antiproliferative and antiangiogenic activity in the MCF-7 cell line and the HCT 116 colon cancer cell line [10][11][12].…”

AC‐1001 H3 CDR peptide induces apoptosis and signs of autophagy in vitro and exhibits antimetastatic activity in a syngeneic melanoma model

“…TEM, considered as the ‘gold standard’ assay to confirm the cell death mechanism 29, was also performed. Alterations in the cell ultrastructure were observed including condensed chromatin, nuclear membrane disintegration, nuclear fragmentation, and plasma membrane blebbing, some well‐known features of apoptosis 28, 30, 31. The human melanoma cell line A2058 was equally affected by AC 1001 H3 at the same cytotoxic concentration showing condensed chromatin and a positive TUNEL assay for DNA degradation.…”

Induction of Apoptosis withMoringa oleiferaFruits in HCT116 Human Colon Cancer Cells Via Intrinsic Pathway

“…The viable cells were stained homogeneously, whereas apoptotic cells which had undergone chromatin condensation and/or nuclear fragmentation were not stained. 17 The HCT116 cells were treated with MOF at different concentrations. Cells were then fixed for 30 min in 100% methanol, washed with PBS, and stained with Hoechst 33258 (2 mg/mL).…”