Suppression of Notch1 and AKT mediated epithelial to mesenchymal transition by Verrucarin J in metastatic colon cancer

Abstract

Epithelial to mesenchymal transition (EMT) in colorectal cancer (CRC) has been attributed to activation of AKT and Notch1 signaling pathways. As EMT corresponds to increased aggressiveness of CRC, approaches that prevent metastasis by targeting AKT/Notch1 pathways are at the forefront of current research paradigms. This study examined the anti-metastatic potential of Verrucarin J (VJ), a small molecule, in CRC cells overexpressing AKT and Notch1. VJ significantly inhibited AKT/HCT 116 cell growth by acting on the AKT/NFκB/Bcl-2 signaling axis and initiated apoptotic signaling as was evident from increased expression of pro-apoptotic markers such as cleaved PARP, cleaved caspase 3, and cleaved caspase 9. Also, VJ inhibited the cell growth in AKT/Notch1-overexpressing CRC cells and abrogated EMT. The down-regulation of AKT and Notch1 signaling was apparent in immunoblot analysis and corresponded with down-regulation of mesenchymal markers including Snail, and β-catenin. Intraperitoneal administration of VJ in control (pCMV/HCT 116) and AKT/HCT 116 mice significantly suppressed AKT-induced tumor growth in a xenograft model. In addition, down-regulation of prosurvival markers as well as AKT and Notch1 was observed in the immunohistochemical analysis of the xenografted tumors. In conclusion, our study substantiates the role of AKT and Notch1 in cell proliferation, angiogenesis, and EMT of CRC cells and demonstrates that VJ may be a viable therapeutic option to counter AKT-induced cell proliferation and tumor outgrowth in CRC.

FABP4 promotes invasion and metastasis of colon cancer by regulating fatty acid transport

“…Phosphorylation of AKT is a key step in the activation of AKT pathway. Studies have shown that hyperactivation of AKT can lead to EMT in in vitro and in vivo CRC models [33]. In our study, we found the level of p-Akt was up-regulated in colon cancer cells with FABP4 overexpression.…”

Recombinant Pyrin Domain Protein Attenuates Airway Inflammation and Alleviates Epithelial-Mesenchymal Transition by Inhibiting Crosstalk Between TGFβ1 and Notch1 Signaling in Chronic Asthmatic Mice

“…As previously mentioned, Notch1 also plays a critical role in the development of EMT (Pal et al, 2018). The Jagged1/Notch1 signaling pathway was therefore also explored.…”

“…NICD can enter the nucleus and combine with the DNA-binding protein RBP-Jκ (also known as CSL) to form a complex that initiates the transcription process of target genes such as Hes1 (LaFoya et al, 2016). Notch1 is involved in the development of EMT in tumor and lung fibrosis models (Pal et al, 2018;Yin et al, 2018). In addition, TGF-β and Notch pathways are synergistically involved in regulating Hes1 expression (Blokzijl et al, 2003).…”

ASR488, a novel small molecule, activates an mRNA binding protein, CPEB1, and inhibits the growth of bladder cancer

“…ting. Annexin V-fluorescein isothiocyanate (FITC) against propidium iodide (PI) assay (FITC Annexin V Apoptosis Detection Kit I, BD Pharminogen) was used for detecting apoptosis as described previously (17). Total protein extracts from TCCSUP cells were prepared with the Mammalian Protein Extraction Reagent (Thermo Scientific) according to the manufacturer's instructions.…”

“…Based on the structure-activity relationship studies focused on the Withaferin A (a dietary compound which exhibits anti-cancer effect against many cancer types) (16)(17)(18)(19)(20)(21)(22) analogs designed in our laboratory, we have identified a novel small molecule, ASR488, designed by protecting-OH group at 4-position of Withaferin A by thiophene-2-carbonyl functionality. ASR-488 demonstrated cell growth arrest in BCa cells and, more importantly, is non-toxic to normal BCa cells.…”