The present study aimed to assess the induction of epithelial-mesenchymal transition (EMT), invasion, and metastasis by the chemokine CXCL9/receptor CXCR3 axis in tongue squamous cell carcinoma (TSCC), unveiling the underlying mechanisms and providing new insights into the prevention and treatment of oral cancer metastasis. The expression levels of CXCL9 and CXCR3 in TSCC tissue specimens were determined by immunohistochemistry, assessing differences between samples with cervical lymph node metastasis and those without. Moreover, protein expression or activity in the TSCC Cal-27 cell line was controlled by neutralizing antibodies, gene transfection, or knock-out. Then, alterations of cell proliferation, migration, invasion, and the cytoskeleton were analyzed by CCK-8, cell scratch, Transwell, and cyto-skeleton staining assays, respectively. Alterations of EMT markers (E-cadherin and vimentin) in Cal-27 cells were detected by immunofluorescence and western blotting. In addition, western blotting was utilized to detect protein expression levels of Akt2, p-Akt2, eIF4E and p-eIF4E, and to explore the regulatory roles and mechanisms of the CXCL9/CXCR3 axis in invasion and metastasis. Significantly increased expression levels of CXCL9 and CXCR3 were detected in tissue specimens with lymph node metastasis compared with those without (P<0.01). Overexpression of CXCL9/CXCR3 in Cal-27 cells resulted in cytoskeleton alterations, decreased E-cadherin expression, increased vimentin levels, enhanced migration and invasion (P<0.05), and increased phosphorylated Akt2 and eIF4E levels (P<0.05). These results revealed that in TSCC, the CXCL9/CXCR3 axis could activate the Akt signaling pathway, with EMT and cytoskeleton rearrangement, promoting invasion and metastasis.

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Objective: To investigate the release of bone morphogenetic protein-2 (BMP-2) and collagen type I proteins (COL1) from different sources ofdemineralized dentine matrix (DDM) and their chemotaxis to mouse osteoprogenitor cells.Methods: The release kinetic of BMP-2 and COL1 was measured from custom-made DDM (CMDDM) and commercially available DDM (CADDM).Using Urist physicochemical method, CMDDM was collected from the extracted teeth in a certified dental clinic. Levels of BMP-2 and COL1 releasedwere measured at days 1, 2, 3, 5, 7, 9, 11, and 13. Next, mouse osteoprogenitor cells, MC3T3-E1, were cultured with a variety of materials as follows:CMDDM, CADDM, Bio-Oss®, and blank control in transwell system. The number of cell migration was determined by crystal violet staining to explorechemotaxis of different DDMs to mouse osteoprogenitor cells.Results: BMP-2 was detected at 588.32 ± 14.53 pg/ml from 5 g of CMDDM. In the release kinetic assay, the concentration of BMP-2 in the CMDDMgroup increased rapidly and peaked at 113.9 pg/ml on day 5, almost four times higher than that of CADDM. The release of COL1 showed similarpattern in both CMDDM and CADDM; however, the amount was significantly higher in the CMDDM group. In cell culture experiment, the number ofmigrated MC3T3-E1 was ranked as the highest in CMDDM, followed by CADDM and Bio-Oss® (p<0.05).Conclusion: CMDDM released BMP-2 and COL1 greater than CADDM, which can induce more osteoblast-like cell migration. These results demonstrateda release kinetic of proteins and osteoinductivity of CMDDM, which supports a benefit of using autogenous bone graft.