ABSTRACT
Adipose-derived stem cells (ADSCs) can differentiate into various cell types and thus have great potential for regenerative medicine. Herein, rat ADSCs were isolated; transduced with lentiviruses expressing Osterix (Osx), a transcriptional factor essential for osteogenesis. Osx overexpression upregulated key osteogenesis-related genes, such as special AT-rich binding protein 2, alkaline phosphatase, osteocalcin, and osteopontin, at both mRNA and protein levels. In addition, mineral nodule formation and alkaline phosphatase activity were enhanced in Osx-overexpressing ADSCs. The expression of dickkopf-related protein 1, a potent Wnt signaling pathway inhibitor, was also increased, whereas that of ß-catenin, an intracellular signal transducer in the Wnt pathway, was decreased. ß-catenin expression was partially recovered by treatment with lithium chloride, a canonical Wnt pathway activator. The Osx-expressing ADSCs were then combined with 3D gelatin-coated porous poly(É-caprolactone) scaffolds with a unique release prolife of entrapped recombinant human vascular endothelial growth factor (VEGF). The controlled release of VEGF promoted osteogenic differentiation capacity in vitro. When the scaffold-ADSC complexes were transplanted into rat calvarial critical-sized defects, more bone formed on the gelatin/VEGF-coated scaffolds than on other scaffold types. Taken together, the results indicate that, Osx-overexpression promotes ADSCs' osteogenesis both in vitro and in vivo, which could be enhanced by release of VEGF.
Subject(s)
Adipose Tissue/metabolism , Osteogenesis/drug effects , Stem Cells/metabolism , Tissue Scaffolds/chemistry , Transcription Factors/biosynthesis , Vascular Endothelial Growth Factor A/pharmacology , Adipose Tissue/cytology , Animals , Delayed-Action Preparations/pharmacology , Female , Rats , Rats, Sprague-Dawley , Stem Cells/cytology , Transcription Factors/geneticsABSTRACT
TRPM2, one member of the transient receptor potential (TRP) protein super-family, is a Ca2+-permeable channel that is activated by oxidative stress and confers susceptibility to cell death. In the human tongue specimens of carcinoma and the tongue carcinoma SCC cell lines, we observed the enhanced expression of TRPM2. By means of the whole-cell electrophysiological recording, the ADPR-induced currents mediated by TRPM2 were recorded in cultured SCC9 cells. Moreover, after H2O2 treatment for 24 hours, the apoptotic number of SCC9 cells was significantly increased. However, the selectively knocked-down TRPM2 with the small interfering RNA technique inhibited the survival and migration of the SCC9 cancer cells, which was independent of the p53-p21 pathway, since the expression of p21 was enhanced after TRPM2 knockdown. Furthermore, the sub-cellular localization of TRPM2 was remarkably different between cancerous and non-cancerous cells. A significant amount of the TRPM2 proteins were located in the nuclei in cancer cells. All these data suggest that TRPM2 is essential for the survival and migration of SCC cancer cells and may be a potential target for the selective treatment of tongue cancer.
Subject(s)
Carcinoma, Squamous Cell/metabolism , Clusterin/metabolism , Mouth Neoplasms/metabolism , Adenosine Diphosphate Ribose/pharmacology , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/pathology , Cell Death/drug effects , Cell Line, Tumor , Cell Movement/drug effects , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Down-Regulation/drug effects , Down-Regulation/genetics , Gene Deletion , Gene Expression Regulation, Neoplastic , Humans , Hydrogen Peroxide/pharmacology , Ion Channel Gating/drug effects , Mouth Neoplasms/genetics , Mouth Neoplasms/pathology , Oxidative Stress/drug effects , Subcellular Fractions/metabolism , Tongue Neoplasms/metabolism , Tongue Neoplasms/pathology , Up-Regulation/drug effects , Up-Regulation/geneticsABSTRACT
BACKGROUND: Both tumor-associated macrophages (TAMs) and the epithelial to mesenchymal transition (EMT) of cancer cells play key roles in promoting tumor progression. However, whether TAMs could induce EMT in the progression of oral squamous cell carcinoma (OSCC) remains undefined. RESULTS: Here we detected the expression of macrophages markers CD68 and CD163, epithelial marker E-cadherin and mesenchymal marker vimentin in 127 OSCC patients by using semi-quantitative immunohistochemistry. CD68 and CD163 expression was not confined to the infiltrating TAMs, but also detected in cancer cells. The high number of CD68-positive macrophages was correlated with poor overall survival. Meanwhile, the expression of CD163 both in macrophages and in cancer cells was associated with poor overall survival and had a significant prognostic impact in OSCC. Importantly, the expression of CD163 in cancer cells had a significant relationship with E-cadherin and vimentin. Furthermore, the incubation of TAMs conditioned medium resulted in a fibroblast-like appearance of cancer cells (HN4, HN6 and SCC9) together with the decreased/increased expression of E-cadherin/ vimentin, which were correlated with the enhanced ability of migration and invasion. CONCLUSIONS: Our results indicate that TAMs could promote the EMT of cancer cells, thereby leading to the progression of oral cancer.
