Matrix metalloproteinase-9 (MMP9) is involved in the TNF-α-induced fusion of human M13SV1-Cre breast epithelial cells and human MDA-MB-435-pFDR1 cancer cells

Abstract

BackgroundIn addition to physiological events such as fertilisation, placentation, osteoclastogenesis, or tissue regeneration/wound healing, cell fusion is involved in pathophysiological conditions such as cancer. Cell fusion, which applies to both the proteins and conditions that induce the merging of two or more cells, is not a fully understood process. Inflammation/pro-inflammatory cytokines might be a positive trigger for cell fusion. Using a Cre-LoxP-based cell fusion assay we demonstrated that the fusion between human M13SV1-Cre breast epithelial cells and human MDA-MB-435-pFDR1 cancer cells was induced by the pro-inflammatory cytokine tumour necrosis factor-α (TNF-α).MethodsThe gene expression profile of the cells in the presence of TNF-α and under normoxic and hypoxic conditions was analysed by cDNA microarray analysis. cDNA microarray data were verified by qPCR, PCR, Western blot and zymography. Quantification of cell fusion events was determined by flow cytometry. Proteins of interest were either blocked or knocked-down using a specific inhibitor, siRNA or a blocking antibody.ResultsThe data showed an up-regulation of various genes, including claudin-1 (CLDN1), ICAM1, CCL2 and MMP9 in M13SV1-Cre and/or MDA-MB-435-pFDR1 cells. Inhibition of these proteins using a blocking ICAM1 antibody, CLDN1 siRNA or an MMP9 inhibitor showed that only the blockage of MMP9 was correlated with a decreased fusion rate of the cells. Likewise, the tetracycline-based antibiotic minocycline, which exhibits anti-inflammatory properties, was also effective in both inhibiting the TNF-α-induced MMP9 expression in M13SV1-Cre cells and blocking the TNF-α-induced fusion frequency of human M13SV1-Cre breast epithelial cells and human MDA-MB-435-pFDR1 cancer cells.ConclusionsThe matrix metalloproteinase-9 (MMP9) is most likely involved in the TNF-α-mediated fusion of human M13SV1-Cre breast epithelial cells and human MDA-MB-435-pFDR1 cancer cells. Likewise, our data indicate that the tetracycline-based antibiotic minocycline might exhibit anti-fusogenic properties because it inhibits a cell fusion-related mechanism.Electronic supplementary materialThe online version of this article (10.1186/s12964-018-0226-1) contains supplementary material, which is available to authorized users.

A network-based approach to identify DNA methylation and its involved molecular pathways in testicular germ cell tumors

“…The Kaplan-Meier analysis results showed that higher levels of MMP9, CSF1R, and PTPRC were related to shorter survival time of TGCT patients. MMP9 is a class of enzymes involved in the degradation of the extracellular matrix, which was up-regulated in tumor tissue samples of multi cancer [59][60][61], such as TGCT [62], breast cancer [63], lung cancer [64]. Moreover, MMP9 overexpression was significantly associated with poor survival of nasopharyngeal carcinoma [65] and glioblastoma [66].…”

Cell Fusion-Mediated Tissue Regeneration as an Inducer of Polyploidy and Aneuploidy

“…Numerous studies further showed that the frequency of cell fusion events was increased upon acute tissue damage or chronic inflammation [7,12,18,32,63,64,[72][73][74][75], which is plausible with regard to efficient BMSC-based and cell fusion-mediated tissue regeneration. BMSCs not only have to be converted into a pro-fusogenic state for subsequent hybridization with target cells but also have to be recruited to the site of tissue damage.…”

“…Osteoclastogenesis [70,74,75], as well as the fusion of cancer cells with endothelial cells [63,76], mesenchymal stem cells [64], or breast epithelial cells [72,73,77] can be induced by TNF-α. Some data revealed that TNF-α could mediate fusion due to induction of MMP-9 expression [73,75], which plays a role in osteoclastogenesis and giant cell formation [75,78]. Hence, it might be assumed that TNF-α could be involved in cell fusion due to induction of pro-fusogenic proteins and/or in an overall conversion of cells into a pro-fusogenic state.…”

Die Rolle der Zellfusion im Tumorkontext. Fakt oder Fiktion?

“…Mehrere Studien deuten auf einen Zusammenhang zwischen Inflammation/ inflammatorische Mediatoren und Zellfusion hin [19,47,61,65,81,90]. Sowohl Johansson et al als auch Davies et al konnten in tierexperimentellen Studien belegen, dass die Fre quenz von Zellfusionsereignissen in Gegenwart von inflammatori schen Konditionen mitunter bis um den Faktor 100 erhöht war im Vergleich zu entsprechenden Kontrollstudien [19,47].…”

“…Sowohl Johansson et al als auch Davies et al konnten in tierexperimentellen Studien belegen, dass die Fre quenz von Zellfusionsereignissen in Gegenwart von inflammatori schen Konditionen mitunter bis um den Faktor 100 erhöht war im Vergleich zu entsprechenden Kontrollstudien [19,47]. Speziell für das pro-inflammatorische Zytokin TNF-α konnte ein pro-fusioge ner Effekt beobachtet werden, was sowohl die Fusion von Makro phagen als auch die Fusion von Tumorzellen mit anderen Zellen be trifft [61,65,81,90]. Auch in chronisch entzündetem Gewebe kommt es zu einer erhöhten Frequenz von Zellfusionsereignissen [47,59,69], da die Zellfusion, wie oben erwähnt, eine Rolle in der Geweberegeneration/ Wundheilung spielt und durch inflammato rische Prozesse begünstigt wird.…”