Bone-invasive Oral Squamous Cell Carcinoma Research Articles

Implications of TP-positive CAFs in the Bone Invasion of Oral Squamous Cell Carcinoma.

Cancer-associated fibroblasts (CAFs) have recently been suggested as critical cellular components of bone invasion in oral squamous cell carcinoma (OSCC). However, the underlying molecular mechanisms and subtypes related to their bone-invasive function are unclear. This study investigated the implications of thymidine phosphorylase (TP)-positive CAFs (TP+CAFs) in OSCC bone invasion. TP expression was determined in 116 patients with OSCC using immunohistochemistry. The influence of TP expression on the biological behavior of CAFs was investigated in vitro. The possible impact of TP+CAFs on bone invasion in OSCC was further evaluated using patient-derived xenograft (PDX) mouse models. In bone-invasive OSCC tissues, TP+CAFs were mainly distributed on the surface of resorbed bone tissue rather than on the tumor side. High levels of TP+CAFs were significantly associated with higher T-stage, bone invasion, and worse overall survival and recurrence-free survival in our study cohort. Recombinant human TP promoted the proliferative and invasive abilities of CAFs and increased matrix metalloproteinase-9 mRNA expression in vitro, related to bone resorption. In the PDX mouse models, TP+CAFs were found in early bone resorption on the surface of resorbed bony tissues. Bone resorption occurred more frequently in the PDX models with TP+CAFs than in those without. TP+CAFs were significantly associated with bone invasion and the prognosis of OSCC. This study provides insights into cellular and molecular targets for the early diagnosis and treatment of bone-invasive OSCC.

Bone invasion by oral squamous cell carcinoma—Is there a link to periodontal disease? A retrospective single center cohort study: 2010–2020

ObjectivesThe aim of this study was to clarify if bone invasion by oral squamous cell carcinoma (OSCC) is associated with periodontal disease (PD), dental status, or tumor location in patients with primary OSCC. Additional overall survival (OS) and recurrence-free survival (RFS) were related to bone invasion and PD. Patients and methodsRetrospectively, 278 patients with OSCC between 2010 and 2020 were included. Uni- and multivariate analyses were performed to assess the association between PD, dental status, tumor location, and the presence of bone invasion by OSCC. Kaplan-Meier curves and logrank tests were used to assess differences in 5-year OS and RFS for the presence of bone invasion and PD. ResultsIn dentate patients (n = 168) periodontal disease was more common in patients with bone-invasive OSCC (90%) compared to patients without OSCC bone invasion (70%), (p = 0.009; OR 3.7 (1.4–9.0)). Floor of mouth (FOM) tumors are more frequent when PD was present (41%). No statistically significant difference was seen in dental status and bone invasion (p = 0.10). Moderate periodontitis was associated with worse OS (55%) compared to severe and no periodontitis (67% and 68% respectively) (p = 0.03; mean survival 42.7 months (SD26.5)). Favorable OS and RFS were significantly associated with no bone invasion (p = 0.047 and 0.035 respectively). ConclusionPD is 3.7 times more common in OSCC with bone invasion and tumor location is related to PD (p < 0.0001). Thus, periodontitis could be a risk factor for bone-invasive OSCC. Elimination of PD might lead to higher OS.

In vitro anticancer activity of a pentacyclic triterpenoid via the mitochondrial pathway in bone-invasive oral squamous cell carcinoma.

Context:Oral cancer is the most dreadful cancer worldwide with a 5-year survival rate of approximately 50%. Anticancer therapies such as chemotherapy and radiotherapy result in severe side effects.Aim:We aimed to evaluate the in vitro anticancer activity of Asiatic acid (AA) on bone-invasive oral squamous cell carcinoma (BHY) cell line.Settings and Design:This was an in vitro laboratory setting.Materials and Methods:BHY cell lines were used for the experiment. Confocal microscopy was used to observe cellular alterations. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the IC50 concentration of AA and flow cytometry to analyze the percentage of cells in each phase of the cell cycle post treatment. Immunoblot assays and semiquantitative reverse transcriptase-polymerase chain reaction (rt-PCR) were used to study the expression level of genes involved.Statistical Analysis Used:Student's t-test and one-way analysis of variance were used for statistical analysis.Results:IC50 concentration of AA was 15.6 μM. On flow cytometry analysis, treatment with 15.6 μM and 31.25 μM of AA for 24 h increased the percentage of cells in the G2/M phase to 45.63% and 53.12%, respectively, compared to 9.62% in control group. Immunoblot analysis and semiquantitative rt-PCR demonstrated an upregulation of p53, cyclin-dependent kinase inhibitors (p21 and p27), caspase-3, caspase-9, cytochrome c and Bax in a time-dependent manner and downregulation of cyclins and anti-apoptotic protein Bcl-2 (**P < 0.05, ***P < 0.001 versus control) post AA treatment.Conclusion:AA induces apoptosis via the mitochondrial-dependent pathway and causes cell cycle arrest at the G2/M phase in BHY cell line.

The Axin2-snail axis promotes bone invasion by activating cancer-associated fibroblasts in oral squamous cell carcinoma

BackgroundIn bone-invasive oral squamous cell carcinoma (OSCC), cancer-associated fibroblasts (CAFs) infiltrate into bony tissue ahead of OSCC cells. In the present study, we aimed to investigate the role of the Axin2-Snail axis in the biological behaviour of CAFs and bone invasion in OSCC.MethodsThe clinicopathological significance of Axin2 and Snail expression was investigated by immunohistochemistry in an OSCC cohort containing 217 tissue samples from patients with long-term follow-up. The influence of the Axin2-Snail axis on the biological behaviour of OSCC cells and CAFs was further investigated both in vitro and in vivo.ResultsAxin2 expression was significantly associated with Snail expression, the desmoplasia status, and bone invasion in patients with OSCC. In multivariate analysis, lymph node metastasis, desmoplasia, Axin2 expression, and Snail expression were independent poor prognostic factors in our cohort. Consistent with these findings, OSCC cells demonstrated attenuated oncogenic activity as well as decreased expression of Snail and various cytokines after Axin2 knockdown in vitro. Among the related cytokines, C-C motif chemokine ligand 5 (CCL5) and interleukin 8 (IL8) demonstrated a strong influence on the biological behaviour of CAFs in vitro. Moreover, both the desmoplastic reaction and osteolytic lesions in the calvaria were predominantly decreased after Axin2 knockdown in OSCC cells in vivo using a BALB/c athymic nude mouse xenograft model.ConclusionsOncogenic activities of the Axin2-Snail axis are not limited to the cancer cells themselves but rather extend to CAFs via regulation of the cytokine-mediated cancer-stromal interaction, with further implications for bone invasion as well as a poor prognosis in OSCC.

Insulin-like growth factor-II mRNA binding protein-3 and podoplanin expression are associated with bone invasion and prognosis in oral squamous cell carcinoma

ObjectiveThis study aimed to evaluate the prognostic implications of insulin-like growth factor-II mRNA binding protein-3 (IMP3) and podoplanin (PDPN) as therapeutic targets against oral squamous cell carcinoma (OSCC) with bone invasion. Study designWe elucidated the correlation of IMP3 and PDPN expression with bone invasion in 160 OSCC tissue specimens, and assessed a mouse calvarium xenograft model using an IMP3- and PDPN-depleted OSCC cell line. ResultsThe retrospective analysis revealed that the expression of IMP3 and PDPN is significantly correlated with T stage, lymph node metastasis, and the overall survival of OSCC patients. In addition, the dual expression of IMP3 and PDPN but not the single expression of either IMP3 or PDPN was associated with bone invasion and the number of osteoclasts in patients with OSCC. In support of these findings, IMP3 or PDPN depletion inhibited the invasive capacity of OSCC cells in a three-dimensional culture system, tumorigenesis, and regional bone destruction in a xenograft mouse model. In addition, IMP3 or PDPN depletion inhibited the expression of interleukin (IL)-6 and IL-8 in OSCC cells, and decreased the expression of receptor activator of NF-κB ligand (RANKL) in xenograft tumor tissues of OSCC. ConclusionsThese results suggest that IMP3 and PDPN may have strong influence on the pathogenesis of OSCC, especially in bone invasion, and may serve as novel therapeutic targets with prognostic implications for bone-invasive OSCC.

An orthotopic and osteolytic model with a newly established oral squamous cell carcinoma cell line

ObjectiveDespite the availability of advanced clinical diagnostic systems, the overall 5-year survival rate for patients with oral squamous cell carcinoma (OSCC) has remained relatively poor over the past 2 decades. Appropriate animal models need to be developed according to the specific purpose of cancer research. However, most of the currently available oral cancer cell lines do not precisely reflect the characteristics of bone-invasive OSCC and cannot serve as suitable tools in the development of new therapeutic reagents against bone-invasive OSCC. Here, we assessed the orthotopic and osteolytic mouse model with newly established OSCC cell line. DesignWe assessed the orthotopic and osteolytic mouse model using the newly established OSCC cells. In addition, the incidences of tumorigenesis and histopathological results were determined. ResultsThe newly established YD-39 cell line grows in a monolayer sheet and has highly invasive. The transplanted YD-39 cells developed stable tumours in the tongues and calvaria region of the nude mice. The tumours in nude mice grafted with YD-39 cells had a high incidence of transplantability in both mouse models tested and a similar morphology to their respective original tumour. Therefore, both animal models might be feasible animal models to assess the efficacy of anti-cancer drugs. ConclusionsThis type of animal model approach might be has the added advantage of potentially accelerating the biological discovery process.

Molecular pathways involved in crosstalk between cancer cells, osteoblasts and osteoclasts in the invasion of bone by oral squamous cell carcinoma

This study investigates whether matrix metalloproteinases (MMPs), specifically MMP-2 and MMP-9, interacting with other molecules important in osteoblast differentiation and osteoclastogenesis, could play important roles in the invasion of bone by oral squamous cell carcinoma (OSCC). Supernatant (conditioned medium, CM) was collected from OSCC cell lines (SCC15 and SCC25), and from cultured osteoblasts (hFOB cell line and a primary culture, OB), and used for indirect co-culture: OSCC cells were treated with CM from osteoblasts and vice versa. Zymogenic activities of MMP-2 and -9, and protein quantities of all molecules studied, were detected by gelatine zymography and Western blotting, respectively. Real-time polymerase chain reaction (PCR) analysed mRNA of these molecules. Targeted molecules were examined by immunohistochemistry in tissue sections of bone-invasive OSCCs. Zymogenic activities of both MMPs were increased in OSCC cells following culture with CM from hFOB: Twist1 protein expression was increased while Runx2 did not alter. The RANKL/OPG ratio, zymogen and protein expression of MMP-9 were increased in hFOB cells cultured with CM from OSCC lines, while zymogen expression of MMP-2 was decreased. Real-time PCR showed generally similar changes in expression of these molecules. All targeted molecules were expressed in invading malignant keratinocytes, and all but OPG were expressed in osteoclasts of clinical samples. Crosstalk between different cell types appears to exist in the invasion of bone by OSCC. Understanding and ultimately interfering with the molecules involved may provide therapeutic approaches to inhibit such bone invasion.

Bone-Invasive Oral Squamous Cell Carcinoma in Cats

Feline oral squamous cell carcinoma (OSCC) is the most common oral tumor in cats. There is no effective treatment, and the average duration of survival after diagnosis is only 2 months. Feline OSCC is frequently associated with osteolysis; however, the mechanisms responsible are unknown. The objective of this study was to characterize the epidemiology and pathology of bone-invasive OSCC in cats and to determine the expression of select bone resorption agonists. In sum, 451 cases of feline OSCC were evaluated. There was no sex or breed predisposition, although there were more intact cats in the OSCC group compared to the control group. Gingiva was the most common site, followed by the sublingual region and tongue. Cats with lingual OSCC were younger (mean, 11.9 years) compared to cats with gingival OSCC (mean, 13.6 years). In addition to osteolysis, there was periosteal new bone formation, osseous metaplasia of tumor stroma, and direct apposition of OSCC to fragments of bone, suggestive of bone-binding behavior. Eighty-two cases were selected for immunohistochemical detection of parathyroid hormone-related protein (PTHrP). Specimens with osteolysis had increased PTHrP expression and nuclear localization, compared to OSCC without osteolysis. Thirty-eight biopsies of OSCC with osteolysis were evaluated for tumor necrosis factor α expression, and only 4 biopsies had such expression in a small proportion of tumor cells. Increased tumor expression of PTHrP and increased localization of PTHrP to the nucleus were associated with osteolysis and may play an important role in bone resorption and tumor invasion in cats with OSCC.

In Vivo and In Vitro Efficacy of Zoledronate for Treating Oral Squamous Cell Carcinoma in Cats

Feline oral squamous cell carcinoma (OSCC) may cause painful bone destruction. Given the local invasiveness and rapid clinical progression of OSCC, conventional therapies are often palliative. In human cancer patients, zoledronate exerts anticancer effects by inhibiting tumor-induced angiogenesis and malignant osteolysis. Zoledronate will exert in vitro and in vivo anti-angiogenic and antiresorptive effects in feline OSCC. Eight cats with OSCC were prospectively treated with zoledronate and conventional treatment modalities. In vitro, zoledronate's effects in modulating soluble vascular endothelial growth factor (VEGF) secretion and receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL) expression were investigated in a feline OSCC cell line (SCCF1). In vivo, basal serum C-telopeptide (CTx) concentrations were compared among normal and OSCC-bearing cats, and the biologic effects of zoledronate administration in cats with naturally occurring OSCC were quantified by serially assessing circulating serum VEGF and CTx concentrations. In vitro, zoledronate concentrations greater than 3 microM reduce soluble VEGF secretion in the SCCF1 cell line. The expression of RANKL in the SCCF1 cell line was also modulated by zoledronate, with low concentrations (3 microM) decreasing but higher concentrations (30 microM) increasing RANKL expression in comparison with untreated cells. In vivo, cats with bone-invasive OSCC had greater serum CTx concentrations in comparison with geriatric, healthy controls. Treatment with zoledronate rapidly decreased circulating serum VEGF and CTx concentrations in cats with spontaneously occurring OSCC. Zoledronate exerts in vitro and in vivo effects that may favor the slowing of tumor growth and pathologic bone turnover associated with OSCC.