Disintegrin Tablysin-15 Suppresses Cancer Hallmarks in Melanoma Cells by Blocking FAK/Akt/ERK and NF-κB Signaling

Abstract

<jats:sec> <jats:title>Background:</jats:title> <jats:p>Integrins are crucial anti-cancer therapy targets. We previously showed that tablysin-15 is an integrin antagonist with its Arg-Gly-Asp motif in a novel structural context.</jats:p> </jats:sec> <jats:sec> <jats:title>Objective:</jats:title> <jats:p>Here we investigated the anti-cancer effects and mechanisms of action of tablysin-15 in melanoma cells.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods:</jats:title> <jats:p>Cell adhesion, competitive binding, cell viability, and ATP chemiluminescence assays were used to analyze the binding of tablysin-15 to αvβ3 integrin and its phenotypic effects. Wound healing, transwells, and zymography were performed to detect motility and matrix metalloproteinase- 2/-9 activities. PARP and caspase-3 cleavage were used as apoptosis assays, while LDH release and flow cytometry were used for necrosis and cell cycle analysis. The expression of mRNAs and proteins of target molecules was measured by qRT-PCR and western blotting, respectively.</jats:p> </jats:sec> <jats:sec> <jats:title>Results:</jats:title> <jats:p>Tablysin-15 dose-dependently inhibited the proliferation, migration, and invasion of M21 cells through integrin αvβ3. The proliferation inhibition caused by tablysin-15 was attributable to G0/G1 phase arrest rather than apoptosis or necrosis. Furthermore, tablysin-15 suppressed MMP-2/- 9 activities and the mRNA expression of MMP-2/-9 and COX-2 but was upregulated TIMP-1 in M21 cells. Meanwhile, tablysin-15 suppressed the expression of cyclin D1/E and CDK 2/6, the phosphorylation of FAK, Akt, and ERK, and nuclear translocation of NF-κB, while increasing the expression of the CDK inhibitor p21waf1/C1. Taken together, tablysin-15 might inhibit melanoma cell metastasis and proliferation by competing with αvβ3 integrin, thereby blocking FAK-associated signaling pathways and nuclear translocation of NF-κB.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions:</jats:title> <jats:p>Tablysin-15 has reliable anti-cancer effects against M21 melanoma cells, suggesting tablysin-15 is a promising anti-tumor drug.</jats:p> </jats:sec>

Upregulation of REV7 correlates with progression of malignant melanoma

“…We found that ERK expression was attenuated in the REV7‐depleted malignant melanoma cells (Figure 4), indicating that REV7 facilitates cell activities by quantitatively regulating the expression of certain signaling molecules. To support this, several reports have demonstrated that the nuclear translocation of ERK1/2, together with PI3K/AKT activation, affects the viability and motility of malignant melanoma cells by regulating the expression of several genes 27–34 . Further research is warranted to clarify intracellular signaling that interact with REV7 expression in malignant melanoma cells.…”

Resveratrol and Its Analogs: Potent Agents to Reverse Epithelial-to-Mesenchymal Transition in Tumors

“…These pro-inflammatory mediators are modulated via the transcription factor NF-kB (113)(114)(115)(116). It has also been reported that focal adhesion kinase (FAK) associated with NF-kB regulates the cancer cells capacity of survival, invasiveness, and metastasis (117)(118)(119)(120). Buhrmann and co-workers showed that RES inhibited the TNF-b-induced EMT as demonstrated by suppressing EMT factors (up-modulating vimentin and slug, down-modulating E-cadherin) in colorectal cancer cells through preventing the FAK/NF-kB activation as proved by downregulation of MMP-9 and C-X-C motif chemokine receptor 4 (CXCR4) which are the NF-kB-dependent of tumor-promoting factors (17,18).…”

Advances in Modern Pharmacology Research of Tabanus

“…Tablysin-15 can inhibit endothelial cell proliferation induced by fibroblast growth factor (FGF) and inhibit angiogenesis in vitro [14] . Deng et al found that Tlysin-15 has anticancer effect on human melanoma cells and binds integrin α V β3 to inhibit proliferation, migration and invasion of tumor cells [15] . Deng et al found that Tlysin-15 binds to αV β3 in vitro to inhibit the proliferation, migration and invasion of breast cancer cell lines [16] .…”