27 research outputs found
Potential role of myeloid-derived suppressor cells in transition from reaction to repair phase of bone healing process
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells with immunosuppressive functions; these cells play a key role in infection, immunization, chronic inflammation, and cancer. Recent studies have reported that immunosuppression plays an important role in the healing process of tissues and that Treg play an important role in fracture healing. MDSCs suppress active T cell proliferation and reduce the severity of arthritis in mice and humans. Together, these findings suggest that MDSCs play a role in bone biotransformation. In the present study, we examined the role of MDSCs in the bone healing process by creating a bone injury at the tibial epiphysis in mice. MDSCs were identified by CD11b and GR1 immunohistochemistry and their role in new bone formation was observed by detection of Runx2 and osteocalcin expression. Significant numbers of MDSCs were observed in transitional areas from the reactionary to repair stages. Interestingly, MDSCs exhibited Runx2 and osteocalcin expression in the transitional area but not in the reactionary area. And at the same area, cllagene-1 and ALP expression level increased in osteoblast progenitor cells. These data is suggesting that MDSCs emerge to suppress inflammation and support new bone formation. Here, we report, for the first time (to our knowledge), the role of MDSCs in the initiation of bone formation. MDSC appeared at the transition from inflammation to bone making and regulates bone healing by suppressing inflammation
Crosstalk between cancer and different cancer stroma subtypes promotes the infiltration of tumorâassociated macrophages into the tumor microenvironment of oral squamous cell carcinoma
Tumorâassociated macrophages (TAMs) are linked to the progression of numerous types of cancer. However, the effects of the tumor microenvironment (TME) of oral squamous cell carcinoma (OSCC), particularly the cancer stroma on TAMs, remains to be elucidated. In the present study, the effects of verrucous SCCâassociated stromal cells (VSCCâSCs), SCCâassociated stromal cells (SCCâSCs) and human dermal fibroblasts (HDFs) on the differentiation, proliferation and migration of macrophages in vitro was assayed using Giemsa staining, and immunofluorescence, MTS and Transwell (migration) assays, respectively. The combined results suggested that both VSCCâSCs and SCCâSCs promoted the differentiation of macrophages into M2 type TAMs, as well as the proliferation and migration of macrophages following crosstalk with HSCâ3 cells in vitro. Moreover, the SCCâSCs exerted a more prominent effect on TAMs than the VSCCâSCs. Immunohistochemical staining was used to examine the expression of CD34, CD45, CD11b and CD163 to assay the effects of VSCCâSCs, SCCâSCs and HDFs on microvessel density (MVD) and the infiltration of CD45(+) monocytes, CD11b(+) TAMs and CD163(+) M2 type macrophages. The results suggested that both VSCCâSCs and SCCâSCs promoted MVD and the infiltration of CD45(+) monocytes, CD11b(+) TAMs and CD163(+) M2 type TAMs into the TME of OSCC following crosstalk with HSCâ3 cells in vivo. The SCCâSCs exerted a more prominent promoting effect than the VSCCâSCs. Finally, the potential genes underlying the differential effects of VSCCâSCs and SCCâSCs on the infiltration of TAMs were investigated using microarray analysis. The results revealed that interleukin 1ÎČ, bone morphogenetic protein 4, interleukin 6 and CâXâC motif chemokine ligand 12 had great potential to mediate the differential effects of VSCCâSCs and SCCâSCs on TAM infiltration. On the whole, the findings presented herein, demonstrate that both VSCCâSCs and SCCâSCs promote the infiltration of TAMs into the TME of OSCC following crosstalk with HSCâ3 cells; the SCCâSCs were found to exert a more prominent promoting effect. This may represent a potential regulatory mechanism for the infiltration of TAMs into the TME of OSCC
Impact of the Stroma on the Biological Characteristics of the Parenchyma in Oral Squamous Cell Carcinoma
Solid tumors consist of the tumor parenchyma and stroma. The standard concept of oncology is that the tumor parenchyma regulates the tumor stroma and promotes tumor progression, and that the tumor parenchyma represents the tumor itself and defines the biological characteristics of the tumor tissue. Thus, the tumor stroma plays a pivotal role in assisting tumor parenchymal growth and invasiveness and is regarded as a supporter of the tumor parenchyma. The tumor parenchyma and stroma interact with each other. However, the influence of the stroma on the parenchyma is not clear. Therefore, in this study, we investigated the effect of the stroma on the parenchyma in oral squamous cell carcinoma (OSCC). We isolated tumor stroma from two types of OSCCs with different invasiveness (endophytic type OSCC (ED-st) and exophytic type OSCC (EX-st)) and examined the effect of the stroma on the parenchyma in terms of proliferation, invasion, and morphology by co-culturing and co-transplanting the OSCC cell line (HSC-2) with the two types of stroma. Both types of stroma were partially positive for alpha-smooth muscle actin. The tumor stroma increased the proliferation and invasion of tumor cells and altered the morphology of tumor cells in vitro and in vivo. ED-st exerted a greater effect on the tumor parenchyma in proliferation and invasion than EX-st. Morphological analysis showed that ED-st changed the morphology of HSC-2 cells to the invasive type of OSCC, and EX-st altered the morphology of HSC-2 cells to verrucous OSCC. This study suggests that the tumor stroma influences the biological characteristics of the parenchyma and that the origin of the stroma is strongly associated with the biological characteristics of the tumor
Significance of cancer stroma for bone destruction in oral squamous cell carcinoma using different cancer stroma subtypes
Stromal cells in the tumor microenvironment (TME) can regulate the progression of numerous types of cancer; however, the bone invasion of oral squamous cell carcinoma (OSCC) has been poorly investigated. In the present study, the effect of verrucous SCCâassociated stromal cells (VSCCâSCs), SCCâassociated stromal cells (SCCâSCs) and human dermal fibroblasts on bone resorption and the activation of HSCâ3 osteoclasts in vivo were examined by hematoxylin and eosin, AE1/3 (panâcytokeratin) and tartrateâresistant acid phosphatase staining. In addition, the expression levels of matrix metalloproteinase (MMP)9, membraneâtype 1 MMP (MT1âMMP), Snail, receptor activator of NFâÎșB ligand (RANKL) and parathyroid hormoneârelated peptide (PTHrP) in the bone invasion regions of HSCâ3 cells were examined by immunohistochemistry. The results suggested that both SCCâSCs and VSCCâSCs promoted bone resorption, the activation of osteoclasts, and the expression levels of MMP9, MT1âMMP, Snail, RANKL and PTHrP. However, SCCâSCs had a more prominent effect compared with VSCCâSCs. Finally, microarray data were used to predict potential genes underlying the differential effects of VSCCâSCs and SCCâSCs on bone invasion in OSCC. The results revealed that IL1B, ICAM1, FOS, CXCL12, INS and NGF may underlie these differential effects. In conclusion, both VSCCâSCs and SCCâSCs may promote bone invasion in OSCC by enhancing the expression levels of RANKL in cancer and stromal cells mediated by PTHrP; however, SCCâSCs had a more prominent effect. These findings may represent a potential regulatory mechanism underlying the bone invasion of OSCC
Secretory Carcinoma of Salivary Gland with High-Grade Histology Arising in Hard Palate: A Case Report
Secretory carcinoma (SC) is a recently described salivary gland tumor reported in the fourth edition of World Health Organization classification of head and neck tumors. SC is characterized by strong S-100 protein, mammaglobin, and vimentin immunoexpression, and harbors a t(12;15)(p13;q25) translocation which leads to ETV6-NTRK3 fusion product. Histologically, SC displays a lobulated growth pattern and is often composed of microcystic, tubular, and solid structures with abundant eosinophilic homogenous or bubbly secretion. SC is generally recognized as low-grade malignancy with low-grade histopathologic features, and metastasis is relatively uncommon. In this case, we described a SC of hard palate that underwent high grade transformation and metastasis to the cervical lymph node in a 54-year-old patient. In addition, this case showed different histological findings between primary lesion and metastasis lesion. Therefore, the diagnosis was confirmed by the presence of ETV6 translocation. Here, we report a case that occurred SC with high-grade transformation in the palate, and a review of the relevant literature is also presented
Investigation of bone invasion and underlying mechanisms of oral cancer using a cell lineâderived xenograft model
The cancer stroma regulates bone invasion in oral squamous cell carcinoma (OSCC). However, data on normal stroma are limited. In the present study, the effects of gingival and periodontal ligament tissueâderived stromal cells (GâSCs and PâSCs, respectively) and human dermal fibroblasts (HDFs) on bone resorption and osteoclast activation were assessed using hematoxylin and eosin and tartrateâresistant acid phosphatase staining in a cell lineâderived xenograft model. The results demonstrated that GâSCs promoted bone invasion and osteoclast activation and inhibited osteoclast proliferation following crosstalk with the human OSCC HSCâ3 cell line, whereas PâSCs inhibited bone resorption and promoted osteoclast proliferation in vitro but had a minimal effect on osteoclast activation both in vitro and in vivo following crosstalk with HSCâ3 cells. Furthermore, the effects of GâSCs, PâSCs and HDFs on protein expression levels of matrix metalloproteinase (MMP)â9, membrane type 1 MMP (MT1âMMP), Snail, parathyroid hormoneârelated peptide (PTHrP) and receptor activator of NFâÎșB ligand (RANKL) in HSCâ3 cells in OSCC bone invasion regions were assessed using immunohistochemistry. The results demonstrated that GâSCs had a more prominent effect on the expression of MMPâ9, MT1âMMP, Snail, PTHrP, and RANKL, whereas PâSCs only promoted RANKL and PTHrP expression and exerted a minimal effect on MMPâ9, MT1âMMP and Snail expression. The potential genes underlying the differential effects of GâSCs and PâSCs on bone invasion in OSCC were evaluated using a microarray, which indicated that cyclinâdependent kinase 1, insulin, aurora kinase A, cyclin B1 and DNA topoisomerase II alpha underlaid these differential effects. Therefore, these results demonstrated that GâSCs promoted bone invasion in OSCC by activating osteoclasts on the bone surface, whereas PâSCs exerted an inhibitory effect. These findings could indicate a potential regulatory mechanism for bone invasion in OSCC
Knockout of MMP3 Weakens Solid Tumor Organoids and Cancer Extracellular Vesicles
The tumor organoid (tumoroid) model in three-dimensional (3D) culture systems has been developed to reflect more closely the in vivo tumors than 2D-cultured tumor cells. Notably, extracellular vesicles (EVs) are efficiently collectible from the culture supernatant of gel-free tumoroids. Matrix metalloproteinase (MMP) 3 is a multi-functional factor playing crucial roles in tumor progression. However, roles of MMP3 within tumor growth and EVs have not unveiled. Here, we investigated the protumorigenic roles of MMP3 on integrities of tumoroids and EVs. We generated MMP3-knockout (KO) cells using the CRISPR/Cas9 system from rapidly metastatic LuM1 tumor cells. Moreover, we established fluorescent cell lines with palmitoylation signal-fused fluorescent proteins (tdTomato and enhanced GFP). Then we confirmed the exchange of EVs between cellular populations and tumoroids. LuM1-tumoroids released large EVs (200-1000 nm) and small EVs (50-200 nm) while the knockout of MMP3 resulted in the additional release of broken EVs from tumoroids. The loss of MMP3 led to a significant reduction in tumoroid size and the development of the necrotic area within tumoroids. MMP3 and CD9 (a category-1 EV marker tetraspanin protein) were significantly down-regulated in MMP3-KO cells and their EV fraction. Moreover, CD63, another member of the tetraspanin family, was significantly reduced only in the EVs fractions of the MMP3-KO cells compared to their counterpart. These weakened phenotypes of MMP3-KO were markedly rescued by the addition of MMP3-rich EVs or conditioned medium (CM) collected from LuM1-tumoroids, which caused a dramatic rise in the expression of MMP3, CD9, and Ki-67 (a marker of proliferating cells) in the MMP3-null/CD9-low tumoroids. Notably, MMP3 enriched in tumoroids-derived EVs and CM deeply penetrated recipient MMP3-KO tumoroids, resulting in a remarkable enlargement of solid tumoroids, while MMP3-null EVs did not. These data demonstrate that EVs can mediate molecular transfer of MMP3, resulting in increasing the proliferation and tumorigenesis, indicating crucial roles of MMP3 in tumor progression
SOD3 Expression in Tumor Stroma Provides the Tumor Vessel Maturity in Oral Squamous Cell Carcinoma
Tumor angiogenesis is one of the hallmarks of solid tumor development. The progressive tumor cells produce the angiogenic factors and promote tumor angiogenesis. However, how the tumor stromal cells influence tumor vascularization is still unclear. In the present study, we evaluated the effects of oral squamous cell carcinoma (OSCC) stromal cells on tumor vascularization. The tumor stromal cells were isolated from two OSCC patients with different subtypes: low invasive verrucous squamous carcinoma (VSCC) and highly invasive squamous cell carcinoma (SCC) and co-xenografted with the human OSCC cell line (HSC-2) on nude mice. In comparison, the CD34+ vessels in HSC-2+VSCC were larger than in HSC-2+SCC. Interestingly, the vessels in the HSC-2+VSCC expressed vascular endothelial cadherin (VE-cadherin), indicating well-formed vascularization. Our microarray data revealed that the expression of extracellular superoxide dismutase, SOD3 mRNA is higher in VSCC stromal cells than in SCC stromal cells. Moreover, we observed that SOD3 colocalized with VE-cadherin on endothelial cells of low invasive stroma xenograft. These data suggested that SOD3 expression in stromal cells may potentially regulate tumor vascularization in OSCC. Thus, our study suggests the potential interest in SOD3-related vascular integrity for a better OSCC therapeutic strategy
Triple knockdown of CDC37, HSP90âalpha and HSP90âbeta diminishes extracellular vesiclesâdriven malignancy events and macrophage M2 polarization in oral cancer
Evidence has been accumulating to indicate that extracellular vesicles (EVs), including exosomes, released by cancer cells can foster tumour progression. The molecular chaperones â CDC37, HSP90α and HSP90ÎČ play key roles in cancer progression including epithelialâmesenchymal transition (EMT), although their contribution to EVsâmediated cellâcell communication in tumour microenvironment has not been thoroughly examined. Here we show that triple depletion of the chaperone trio attenuates numerous cancer malignancy events exerted through EV release. Metastatic oral cancerâderived EVs (MEV) were enriched with HSP90α HSP90ÎČ and cancerâinitiating cell marker CD326/EpCAM. Depletion of these chaperones individually induced compensatory increases in the other chaperones, whereas triple siRNA targeting of these molecules markedly diminished the levels of the chaperone trio and attenuated EMT. MEV were potent agents in initiating EMT in normal epithelial cells, a process that was attenuated by the triple chaperone depletion. The migration, invasion, and in vitro tumour initiation of oral cancer cells were significantly promoted by MEV, while triple depletion of CDC37/HSP90α/ÎČ reversed these MEVâdriven malignancy events. In metastatic oral cancer patientâderived tumours, HSP90ÎČ was significantly accumulated in infiltrating tumourâassociated macrophages (TAM) as compared to lower grade oral cancer cases. HSP90âenriched MEVâinduced TAM polarization to an M2 phenotype, a transition known to support cancer progression, whereas the triple chaperone depletion attenuated this effect. Mechanistically, the triple chaperone depletion in metastatic oral cancer cells effectively reduced MEV transmission into macrophages. Hence, siRNAâmediated knockdown of the chaperone trio (CDC37/HSP90α/HSP90ÎČ) could potentially be a novel therapeutic strategy to attenuate several EVâdriven malignancy events in the tumour microenvironment