The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis

Abstract

Though Brassinin is known to have antiangiogenic, anti-inflammatory, and antitumor effects in colon, prostate, breast, lung, and liver cancers, the underlying antitumor mechanism of Brassinin is not fully understood so far. Hence, in the current study, the apoptotic mechanism of Brassinin was explored in prostate cancer. Herein, Brassinin significantly increased the cytotoxicity and reduced the expressions of pro-Poly ADP-ribose polymerase (PARP), pro-caspase 3, and B-cell lymphoma 2 (Bcl-2) in PC-3 cells compared to DU145 and LNCaP cells. Consistently, Brassinin reduced the number of colonies and increased the sub-G1 population and terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL)-positive cells in the PC-3 cells. Of note, Brassinin suppressed the expressions of pyruvate kinase-M2 (PKM2), glucose transporter 1 (GLUT1), hexokinase 2 (HK2), and lactate dehydrogenase (LDH) as glycolytic proteins in the PC-3 cells. Furthermore, Brassinin significantly reduced the expressions of SIRT1, c-Myc, and β-catenin in the PC-3 cells and also disrupted the binding of SIRT1 with β-catenin, along with a protein–protein interaction (PPI) score of 0.879 and spearman’s correlation coefficient of 0.47 being observed between SIRT1 and β-catenin. Of note, Brassinin significantly increased the reactive oxygen species (ROS) generation in the PC-3 cells. Conversely, ROS scavenger NAC reversed the ability of Brassinin to attenuate pro-PARP, pro-Caspase3, SIRT1, and β-catenin in the PC-3 cells. Taken together, these findings support evidence that Brassinin induces apoptosis via the ROS-mediated inhibition of SIRT1, c-Myc, β-catenin, and glycolysis proteins as a potent anticancer candidate.

Exploring the Antiproliferative and Modulatory Effects of 1-Methoxyisobrassinin on Ovarian Cancer Cells: Insights into Cell Cycle Regulation, Apoptosis, Autophagy, and Its Interactions with NAC

“…In addition, markers of apoptosis, such as phosphatidylserine externalization, revealed a notable reduction in viable cells and an increase in apoptotic and necrotic populations following exposure to MB-591. These results are in line with published studies where several other natural indole phytoalexins or their synthetic derivatives induced apoptosis in various cancer cells by downregulating pro-PARP, activating different caspases, decreasing MMP, and increasing the concentration of ROS [12,17,18,[54][55][56].…”

Anticancer Potential of Indole Phytoalexins and Their Analogues

“…Moreover, brassinin triggered ROS production in PC-3 cells, significantly contributing to its antitumor effects. The use of the ROS scavenger N-acetyl-L-cysteine (NAC) reversed brassinin's impact on the expressions of pro-PARP, pro-caspase 3, SIRT1, and β-catenin, highlighting the crucial role of ROS in brassinin's antitumor properties [41]. Similar results were observed in the study by Hong et al in 2021, where NAC effectively inhibited brassinin-induced ROS production in Huh7 cells (human hepatocellular carcinoma cells with mutated p53).…”

“…Brassinin demonstrates anticancer potential in various cancers such as myelogenous leukemia [39], colon [40], prostate [41], breast [42], lung [43], and nasopharyngeal [44], hepatocellular carcinoma [17]. It shows great potential as a prospective anticancer agent due to its ability to induce apoptosis, inhibit angiogenesis, and modulate crucial signaling pathways involved in cancer development and progression, such as the NF-κB pathway, Janus kinase/signal transducer and activator of transcription 3 (JAK-STAT3), pathway and Phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway.…”