Inhibition of αvβ3 integrin induces loss of cell directionality of oral squamous carcinoma cells (OSCC)

Abstract

The connective tissue formed by extracellular matrix (ECM) rich in fibronectin and collagen consists a barrier that cancer cells have to overpass to reach blood vessels and then a metastatic site. Cell adhesion to fibronectin is mediated by αvβ3 and α5β1 integrins through an RGD motif present in this ECM protein, thus making these receptors key targets for cell migration studies. Here we investigated the effect of an RGD disintegrin, DisBa-01, on the migration of human fibroblasts (BJ) and oral squamous cancer cells (OSCC, SCC25) on a fibronectin-rich environment. Time-lapse images were acquired on fibronectin-coated glass-bottomed dishes. Migration speed and directionality analysis indicated that OSCC cells, but not fibroblasts, showed significant decrease in both parameters in the presence of DisBa-01 (1μM and 2μM). Integrin expression levels of the α5, αv and β3 subunits were similar in both cell lines, while β1 subunit is present in lower levels on the cancer cells. Next, we examined whether the effects of DisBa-01 were related to changes in adhesion properties by using paxillin immunostaining and total internal reflection fluorescence TIRF microscopy. OSCCs in the presence of DisBa-01 showed increased adhesion sizes and number of maturing adhesion. The same parameters were analyzed usingβ3-GFP overexpressing cells and showed that β3 overexpression restored cell migration velocity and the number of maturing adhesion that were altered by DisBa-01. Surface plasmon resonance analysis showed that DisBa-01 has 100x higher affinity for αvβ3 integrin than forα5β1 integrin. In conclusion, our results suggest that the αvβ3 integrin is the main receptor involved in cell directionality and its blockage may be an interesting alternative against metastasis.

Matrix stiffness mechanically conditions EMT and migratory behavior of oral squamous cell carcinoma

“…A stiffer niche enhances nuclear translocation of YAP (Taubenberger et al, 2016) and signals (Calvo et al, 2013) that promote focal adhesion assembly (Nardone et al, 2017), but weaker Inv H /E:N L cell-matrix adhesion might indicate that invasive OSCC cells migrate together and slower than cells of other tumors, whose preferred mode of migration is as single cells (Nguyen-Ngoc et al, 2012). However, unlike Nguyen-Ngoc and co-workers, we have only explored OSCC migration on type I collagen substrates, but ECM composition directly modulates migration (Ramos Gde et al, 2016) and how processive cells can be (Montenegro et al, 2017). Thus, our observation of faster collective migration could be limited by ECM ligand, whichalthough type I collagen is the dominant ligand in tumor-adjacent stroma (Pickup et al, 2014) could nonetheless alter the ratio of cells that migrate individually compared with those migrating collectively.…”

High αv Integrin Level of Cancer Cells Is Associated with Development of Brain Metastasis in Athymic Rats

“…MicroRNA-124 inhibited hepatocarcinoma cell invasion and migration by suppressing αv integrin (49). Inhibition of αvβ3 integrin by RGD disintegrin also induces loss of cell directionality and reduction of migration speed in oral squamous carcinoma cells (50). Cellular integrin regulation, trafficking and recycling is extremely complicated processes highly dependent upon the activation and ligand bound state of integrin (28, 51–53).…”

Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

“…RGD is usually recognized by both α5β1 and αvβ3 integrins, however, these two integrins play divergent roles in cell adhesion and migration. Fibronectin adhesion by α5β1 integrin results in highly dynamic thin cell protrusions in multiple directions while adhesion to αvβ3 integrin results in a single large lamellipodium with more static adhesions at the leading edge [13][14][15]. In addition to the RGD motif, integrin and ECM conformations are crucial to their interaction indicating a complex mechanism [16,17].…”

“…Integrin αvβ3 is highly expressed in aggressive cancers, which is related to increase of tumor cell migration, adhesion and invasion during tumor progression [13,[18][19][20][21][22]. Since integrin inhibition blocks cell migration, these receptors were considered a valuable target on cancer research [16,[23][24][25]. However, cilengitide, a cyclic RGD-containing peptide that antagonizes αvβ3 and α5β1 integrins at nanomolar ranges had promising pre-clinical results but it was ineffective in phase I clinical outcomes [16].…”