Efficacy of melatonin, IL-25 and siIL-17B in tumorigenesis-associated properties of breast cancer cell lines

Abstract

Mammary tumorigenesis can be modulated by melatonin, which has oncostatic action mediated by multiple mechanisms, including the inhibition of the activity of transcription factors such as NF-κB and modulation of interleukins (ILs) expression. IL-25 is an active cytokine that induces apoptosis in tumor cells due to differential expression of its receptor (IL-17RB). IL-17B competes with IL-25 for binding to IL-17RB in tumor cells, promoting tumorigenesis. This study purpose is to address the possibility of engaging IL-25/IL-17RB signaling to enhance the effect of melatonin on breast cancer cells. Breast cancer cell lines were cultured monolayers and 3D structures and treated with melatonin, IL-25, siIL-17B, each alone or in combination. Cell viability, gene and protein expression of caspase-3, cleaved caspase-3 and VEGF-A were performed by qPCR and immunofluorescence. In addition, an apoptosis membrane array was performed in metastatic cells. Treatments with melatonin and IL-25 significantly reduced tumor cells viability at 1mM and 1ng/mL, respectively, but did not alter cell viability of a non-tumorigenic epithelial cell line (MCF-10A). All treatments, alone and combined, significantly increased cleaved caspase-3 in tumor cells grown as monolayers and 3D structures (p<0.05). Semi-quantitative analysis of apoptosis pathway proteins showed an increase of CYTO-C, DR6, IGFBP-3, IGFBP-5, IGFPB-6, IGF-1, IGF-1R, Livin, P21, P53, TNFRII, XIAP and hTRA proteins and reduction of caspase-3 (p<0.05) after melatonin treatment. All treatments reduced VEGF-A protein expression in tumor cells (p<0.05). Our results suggest therapeutic potential, with oncostatic effectiveness, pro-apoptotic and anti-angiogenic properties for melatonin and IL-25-driven signaling in breast cancer cells.

Melatonin restrains angiogenic factors in triple-negative breast cancer by targeting miR-152-3p: In vivo and in vitro studies

“…It has already been demonstrated by our group that melatonin regulates angiogenic proteins in the MDA-MB-231 cell line and in the co-culture with cancer-associated fibroblasts [28]. This hormone also regulates angiogenesis under hypoxia in MCF-7 and MDA-MB-231 cells [29] and decreases the expression of genes related to this process in MDA-MB-231 and MCF-7 cells in three-dimensional culture [30]. Thus, genes related to angiogenesis can be regulated by various mechanisms and molecules, including miRNAs [31] and melatonin [32].…”

The emergence of melatonin in oncology: Focus on colorectal cancer

“…Moreover, melatonin lacks the undesirable side effects of many synthetic molecules. Likewise, the strong link of NF‐κβ with CRC, and the implication of this transcription factor in the antiangiogenic and anti‐inflammatory responses of melatonin, merit attention to determine whether in CRC cells the indoleamine produces a coordinated engagement of angiogenic and inflammatory proteins with the aim to enhance oncostasis, as has already been demonstrated in breast cancer cells …”

“…Likewise, the strong link of NF-κβ with CRC, and the implication of this transcription factor in the antiangiogenic and anti-inflammatory responses of melatonin, merit attention to determine whether in CRC cells the indoleamine produces a coordinated engagement of angiogenic and inflammatory proteins with the aim to enhance oncostasis, as has already been demonstrated in breast cancer cells. 208 Recent discoveries show that another CRC-stimulating circuit activated by the ET-1, together with its endothelin receptor A (ET-1/ET-AR axis), is the Yes-associated protein (YAP) and PDZ-binding motif (TAZ) signaling system. 209 YAP/TAZ are two structural components of the β-catenin destruction complex that regulate the Wnt signaling pathway; they are also downstream transcriptional coactivators of the highly conserved Hippo pathway.…”

Mechanisms Underlying Tumor Suppressive Properties of Melatonin

“…Synergistic cytotoxicity of melatonin and anticancer drugs toward leukemia lymphocytes but not normal lymphocytes has been described [61]. Furthermore, viability of malignant cells but not their normal counterparts is reduced by melatonin [62]. SIRT1 is a member of the sirtuin family of proteins that plays a role in chromatin remodeling and gene expression.…”

“…Inhibition of expression of vascular endothelial growth factor is a central part of this targeting [105]. Melatonin-promoted inhibition of inflammatory events also plays a role in retarding blood vessel proliferation [106] and melatonin can simultaneously reduce content of angiogenic and inflammatory proteins in a mammary tumor cell line [62]. In hepatocellular carcinoma cells, prazosin, an MT3 receptor antagonist, can strongly upregulate the IL-6 gene, involved in inflammatory events and in promoting angiogenesis [107].…”