Programmed Cell Death Alterations Mediated by Synthetic Indole Chalcone Resulted in Cell Cycle Arrest, DNA Damage, Apoptosis and Signaling Pathway Modulations in Breast Cancer Model

Abstract

Although new chemotherapy significantly increased the survival of breast cancer (BC) patients, the use of these drugs is often associated with serious toxicity. The discovery of novel anticancer agents for BC therapy is expected. This study was conducted to explore the antiproliferative effect of newly synthesized indole chalcone derivative ZK-CH-11d on human BC cell lines. MTT screening, flow cytometry, Western blot, and fluorescence microscopy were used to evaluate the mode of cell death. ZK-CH-11d significantly suppressed the proliferation of BC cells with minimal effect against non-cancer cells. This effect was associated with cell cycle arrest at the G2/M phase and apoptosis induction. Apoptosis was associated with cytochrome c release, increased activity of caspase 3 and caspase 7, PARP cleavage, reduced mitochondrial membrane potential, and activation of the DNA damage response system. Furthermore, our study demonstrated that ZK-CH-11d increased the AMPK phosphorylation with simultaneous inhibition of the PI3K/Akt/mTOR pathway indicating autophagy initiation. However, chloroquine, an autophagy inhibitor, significantly potentiated the cytotoxic effect of ZK-CH-11d in MDA-MB-231 cells indicating that autophagy is not principally involved in the antiproliferative effect of ZK-CH-11d. Taking together the results from our experiments, we assume that autophagy was activated as a defense mechanism in treated cells trying to escape from chalcone-induced harmful effects.

Chalcone-Acridine Hybrid Suppresses Melanoma Cell Progression via G2/M Cell Cycle Arrest, DNA Damage, Apoptosis, and Modulation of MAP Kinases Activity

“…In the present study, we documented increased JNK and p38 phosphorylation in both 1C -treated melanoma cells. Our results agreed with previous studies where the association between JNK and p38 phosphorylation and chalcone-induced apoptosis was documented [ 40 , 41 , 98 , 99 ]. Surprisingly, we also found the activation (i.e., phosphorylation) of Erk1/2 which, generally, prevents the apoptosis via either downregulation of proapoptotic or the upregulation of antiapoptotic proteins [ 93 ].…”

“…It has been documented that an increased Bax/Bcl-xL ratio supports apoptosis due to cytochrome c release with subsequent activation of caspases, as mentioned earlier [ 80 ]. Similar results were obtained with either synthetic or natural chalcones in different cancer cells [ 41 , 81 ]. Furthermore, we observed the increased phosphorylation of antiapoptotic Bcl-2 and proapoptotic Bad proteins.…”

“…In our recent article [ 41 ], we mentioned pleiotropic mechanisms of the antiproliferative effect of chalcones in in vitro cancer models. Among others, chalcones caused cell cycle arrest, mostly at the G2/M phase [ 36 , 65 , 66 ].…”

“…Due to the structural heterogeneity, chalcones are useful templates for the development of novel active compounds with more convenient biological activities [ 34 , 35 ]. In the last decade we documented the antiproliferative effect of several chalcone derivatives such as acridine hybrids [ 36 , 37 , 38 ], indole hybrids [ 39 , 40 , 41 ] or cyclic chalcone analogues [ 42 , 43 , 44 ] using different in vitro cancer models such as breast, colorectal or cervix cancers. Furthermore, both the antiproliferative and anticancer effects of chalcones have also been documented using melanoma cancer cells or melanoma xenografts [ 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 ].…”

Chalcone-3 Inhibits the Proliferation of Human Breast Cancer MDA-MB-231 Cell Line

“…Furthermore, this process led to the reduction of cyclin-D1 and G2/M arrest. MDA-MB-231 cells treated with synthetic chalcone (ZK-CH-11d) for 24 h were also shown to cause G2/M cell cycle arrest by significantly downregulating phosphorylated cyclin-B1, cyclin-D1, cdc2, Rb and altering the expression of α, α1c tubulin, and β tubulin with a maximum of 72 h after treatment (Michalkova et al, 2022). The same result was reported by Elkhalifa et al, (2020) These consequently led to cell cycle arrest at the G2/M phases (Oh and Seo, 2017).…”