Acetazolamide triggers death inducing autophagy in T‐47D breast cancer cells

Abstract

The inhibitory effects of acetazolamide on the growth and proliferation of epithelial breast cancer cells (T-47D) were investigated. Analysis of morphological changes indicated little apoptosis in T-47D cells incubated with acetazolamide, according to data from flow cytometry, DNA laddering, and expression of AIF. However, an increase in caspase-3 activity was detected in cells. This was concomitant with an increase in DFF45/DFF40 ratio leading to inhibition of caspase-3 activity, DNA fragmentation and progression of apoptosis. Flow cytometry also confirmed that acetazolamide had no significant effect on cell cycle progression. These results are consistent with lack of change in the expression of cell cycle regulatory proteins p21, p27, cdc2 and cyclinD1. Increased expression of ATG5, p53 and DRAM, along with an increase in BCLN1/Bcl-2 ratio, indicated that acetazolamide inhibited the proliferation of T-47D cells by inducing autophagy. Increased expression of PTEN, along with decreased expression of Akt1, also showed that acetazolamide treatment resulted in death inducing autophagy. Collectively the results indicate that autophagy is an adequate mechanism mediating the anti-cancer effects of acetazolamide in T-47D cells through engagement of p53/DRAM pathway and attenuation of Akt survival signalling.

Simultaneous Targeting of Bladder Tumor Growth, Survival, and Epithelial-to-Mesenchymal Transition with a Novel Therapeutic Combination of Acetazolamide (AZ) and Sulforaphane (SFN)

“…Both AZ and SFN have been reported to modulate individually Akt signalling in T-47D breast cancer cells and colon adenocarcinoma Caco2 cell [41,31]. Here we found that downregulation of the Akt signalling protein was correlated with induced expression of cell cycle inhibitors p21, p27, reduced cyclin D1, PARP, and caspase-3 cleavage, further supporting the targeting of the Akt signalling pathway by the AZ+SFN combination.…”

“…Using our in vitro and in vivo models, we showed that growth of the tumors was significantly reduced by the AZ+SFN combination, although, the single agent SFN alone still showed an appreciable strong antitumor efficacy in the xenografted mice. Both AZ and SFN have been shown to induce apoptosis in human Caco2 and breast cancer cells and decrease pro-survival Akt phosphorylation [41,31]. Furthermore, SFN alone showed a downregulation tendency of both basal and p-Akt expression in ovarian cancer cells [45].…”

Betulinyl Sulfamates as Anticancer Agents and Radiosensitizers in Human Breast Cancer Cells

“…Moreover, apoptosis induction was detected in caspase 3 defective MCF7 cells ( Figure 2 ). Recently, caspase-independent forms of BA-induced cell death were discussed [ 31 ]. Consistent with this result, Lou et al suggested caspase-independent apoptosis induction by CAIX shRNA [ 29 ].…”

Augmentation of the cytotoxic effects of zinc oxide nanoparticles by MTCP conjugation: Non-canonical apoptosis and autophagy induction in human adenocarcinoma breast cancer cell lines

“…When AIF is released into the cytosol, it triggers caspase-independent apoptotic death via DNA fragmentation (Candé et al, 2002). In fact, in MDA-MB-468 treated cells, in addition to the increased expression level of bcl2 and its effects on the calcium-dependent lysosomal death, the caspase-independent apoptotic death, induced by AIF, (AIF dependent apoptosis) was also another auxiliary mechanism that helps calcium ions to induce cell death (Mohammadpour et al, 2014). …”