Oral squamous carcinoma cells secrete RANKL directly supporting osteolytic bone loss

Objective: Local invasion of bone is a frequent complication of oral squamous cell carcinoma (OSCC). Development of these osteolytic lesions is mediated by osteoclasts. Receptor activation of NF-kappa B ligand (RANKL) signaling, counteracted by osteoprotegerin (OPG), regulates osteoclastogenesis. Previous studies in rodent models have demonstrated that inhibition of RANKL decreases tumor growth and lesions within bone. However, the contributory role of OSCC cells to this disease process has yet to be defined.Methods: RANKL expression was assessed in a panel of OSCC cell lines by qPCR, flow cytometry, and ELISA. Induction of osteoclastogenesis was assessed by co-culture with macrophages or with OSCC-derived conditioned medium. In an animal model of bone invasion, nude mice were injected intratibially with UMSCC-11B cells expressing a RANKL luciferase promoter to detect tumor-derived RANKL activity. Osteolytic lesions were analyzed by X-ray, micro-CT, and histological methods. RANKL expression was assessed in human OSCC tissues by immunohistochemistry.Results: We demonstrated that OSCCs express varied levels of all RANKL isoforms, both membrane-bound and soluble RANKL. Both co-culture and treatment with OSCC-conditioned media induced osteoclastogenesis. In mice, we demonstrated human RANKL promoter activity during bone invasion. Over the course of the experiment, animals suffered osteolytic lesions as RANKL-driven luciferase expression increased with time. After 8 weeks, human-derived RANKL was detected in areas of bone resorption by immunohistochemistry. Similar epithelial RANKL expression was detected in human OSCC tissues.Conclusion: These data demonstrate the ability of OSCCs to produce RANKL, directly altering the tumor microenvironment to increase osteoclastogenesis and mediate local bone invasion. (C) 2012 Elsevier Ltd. All rights reserved.

IB4(+) and TRPV1(+) sensory neurons mediate pain but not proliferation in a mouse model of squamous cell carcinoma

Background: Cancer pain severely limits function and significantly reduces quality of life. Subtypes of sensory neurons involved in cancer pain and proliferation are not clear.Methods: We produced a cancer model by inoculating human oral squamous cell carcinoma (SCC) cells into the hind paw of athymic mice. We quantified mechanical and thermal nociception using the paw withdrawal assays. Neurotoxins isolectin B4-saporin (IB4-SAP), or capsaicin was injected intrathecally to selectively ablate IB4(+) neurons or TRPV1(+) neurons, respectively. JNJ-17203212, a TRPV1 antagonist, was also injected intrathecally. TRPV1 protein expression in the spinal cord was quantified with western blot. Paw volume was measured by a plethysmometer and was used as an index for tumor size. Ki-67 immunostaining in mouse paw sections was performed to evaluate cancer proliferation in situ.Results: We showed that mice with SCC exhibited both mechanical and thermal hypersensitivity. Selective ablation of IB4(+) neurons by IB4-SAP decreased mechanical allodynia in mice with SCC. Selective ablation of TRPV1(+) neurons by intrathecal capsaicin injection, or TRPV1 antagonism by JNJ-17203212 in the IB4-SAP treated mice completely reversed SCC-induced thermal hyperalgesia, without affecting mechanical allodynia. Furthermore, TRPV1 protein expression was increased in the spinal cord of SCC mice compared to normal mice. Neither removal of IB4(+) or TRPV1(+) neurons affected SCC proliferation.Conclusions: We show in a mouse model that IB4(+) neurons play an important role in cancer-induced mechanical allodynia, while TRPV1 mediates cancer-induced thermal hyperalgesia. Characterization of the sensory fiber subtypes responsible for cancer pain could lead to the development of targeted therapeutics.

Efeitos toxicogenéticos e toxicogenômicos do Isotiocianato de Alila (óleo de mostarda) em linhagens celulares de carcinoma de bexiga

Pós-graduação em Patologia - FMB

Metástases de carcinoma de mama em cadelas: avaliação da técnica de linfonodo sentinela

Pós-graduação em Medicina Veterinária - FMVZ

Estudo da viabilidade do tratamento de pacientes portadores de carcinoma epidermóide de lábio(T1nomo) por meio da radiação laser de dióxido de carbono (CO2)

Pós-graduação em Odontologia - ICT

Diagnóstico, tratamento e seguimento do carcinoma medular de tireoide: recomendações do Departamento de Tireoide da Sociedade Brasileira de Endocrinologia e Metabologia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Invasive head and neck cutaneous squamous cell carcinoma: clinical and histopathological characteristics, frequency of local recurrence and metastasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fatores prognósticos e análise de sobrevida de pacientes diagnosticados com carcinoma espinocelular nos 10 primeiros anos do Centro de Oncologia Bucal da Unesp, Campus de Araçatuba

Objetivo: Estimar as probabilidades acumuladas de sobrevida dos pacientes diagnosticados com carcinoma espinocelular nos 10 primeiros anos do Centro de Oncologia Bucal da UNESP, Campus de Araçatuba, de 1991 a 2000, observadas até 2005, estabelecendo os possíveis fatores prognósticos significativos para o óbito. Méttodo: A análise de sobrevida foi realizada em uma coorte de 280 pacientes com carcinoma espinocelular, no Centro de Oncologia Bucal da Faculdade de Odontologia de Araçatuba, UNESP, entre 1991 e 2000. Para avaliar a associação entre as variáveis independentes e o óbito, realizou-se o teste Log Rank. A probabilidade do teste com p-valor menor que 0,25 ficou estabelecida para a inclusão das covariáveis no processo de ajustamento do modelo. A sobrevida foi estimada pelo método de produto limite de Kaplan-Meier. Os fatores prognósticos foram estimados pelo modelo de riscos proporcionais de Cox, calculando-se razão da função de risco (HR). A análise de resíduo foi realizada para verificar o ajuste do modelo. Resultados: As taxas de probabilidades acumuladas de sobrevida de 280 pacientes, para os casos em estádio IV, foram, 56,74%, 32,13%, 23,71% e 20,57%, respectivamente, até 1, 2, 3 e 5 anos após o diagnóstico. Pacientes no estádio I apresentaram sobrevida em 5 anos de 81,73%. O estadiamento clínico da doença no diagnóstico foi o único fator prognóstico definido no processo de ajuste de modelo. A estimativa da razão da função de riscos de morrer em pacientes diagnosticados no estádio III (HR=3,3), é praticamente três vezes o risco daqueles em estádio I; da mesma forma, o risco de morrer dos diagnosticados em estádio IV (HR=6,17) é cerca de seis vezes ao daqueles em estádio I. Conclusões: A covariável que permaneceu no modelo final foi estadiamento clínico no momento do diagnóstico, sendo, pois, o único fator prognóstico.

Stromal myofibroblasts in squamous cell carcinoma of the tongue in young patients - a multicenter collaborative study

Objective. The purpose of this study was to evaluate the presence of myofibroblasts, frequently associated with a more aggressive neoplastic behavior, in oral tongue squamous cell carcinoma (TSCC) of young patients and to compare with the distribution observed in older patients.Study Design. Tumor samples from 29 patients younger than 40 years old affected by TSCC were retrieved and investigated for the presence of stromal myofibroblasts by immunohistochemical reactions against a smooth muscle actin, and the results obtained were compared to TSCC cases affecting older patients.Results. No positive reaction could be found in the stromal areas devoid of neoplastic tissue, whereas myofibroblasts were present in 58.6% of the lesions in young patients and in 75.9% of the older ones. No significant difference was found when comparing the invasive front and the overall stroma of both groups, and no correlation could be obtained with stromal a smooth muscle actin expression, higher tumor grades or clinical stage (P > .05).Conclusion. There was no significant difference between the presence of stromal myofibroblasts of TSCC affecting young and old individuals.

GPC3 reduces cell proliferation in renal carcinoma cell lines

Background: Glypican 3 (GPC3) is a member of the family of glypican heparan sulfate proteoglycans (HSPGs). The GPC3 gene may play a role in controlling cell migration, negatively regulating cell growth and inducing apoptosis. GPC3 is downregulated in several cancers, which can result in uncontrolled cell growth and can also contribute to the malignant phenotype of some tumors. The purpose of this study was to analyze the mechanism of action of the GPC3 gene in clear cell renal cell carcinoma.Methods: Five clear cell renal cell carcinoma cell lines and carcinoma samples were used to analyze GPC3 mRNA expression (qRT-PCR). Then, representative cell lines, one primary renal carcinoma (786-O) and one metastatic renal carcinoma (ACHN), were chosen to carry out functional studies. We constructed a GPC3 expression vector and transfected the renal carcinoma cell lines, 786-O and ACHN. GPC3 overexpression was analyzed using qRT-PCR and immunocytochemistry. We evaluated cell proliferation using MTT and colony formation assays. Flow cytometry was used to evaluate apoptosis and perform cell cycle analyses.Results: We observed that GPC3 is downregulated in clear cell renal cell carcinoma samples and cell lines compared with normal renal samples. GPC3 mRNA expression and protein levels in 786-O and ACHN cell lines increased after transfection with the GPC3 expression construct, and the cell proliferation rate decreased in both cell lines following overexpression of GPC3. Further, apoptosis was not induced in the renal cell carcinoma cell lines overexpressing GPC3, and there was an increase in the cell population during the G1 phase in the cell cycle.Conclusion: We suggest that the GPC3 gene reduces the rate of cell proliferation through cell cycle arrest during the G1 phase in renal cell carcinoma.

Wavelet-based algorithm to the evaluation of contrasted hepatocellular carcinoma in CT-images after transarterial chemoembolization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Minimally Invasive Selective Neck Dissection: A Prospective Study of Endoscopically Assisted Dissection via a Small Submandibular Approach in cT(1-2)N(0) Oral Squamous Cell Carcinoma

Selective neck dissection (SND) in clinical N-0 (cN(0)) cases of oral squamous cell carcinoma (SCC) has been performed by surgeons using a retroauricular or modified facelift approach with robotic or endoscopic assistance. However, these procedures provide cosmetic satisfaction at the cost of possible maximal invasiveness. In this prospective study, we introduced and evaluated the feasibility as well as surgical invasiveness and cosmetic outcome of endoscopically-assisted SND via a small submandibular approach.Forty-four patients with cT(1-2)N(0) oral SCC (OSCC) were randomly divided into two groups of endoscopically-assisted SND and conventional SND. Perioperative and postoperative outcomes of patients were evaluated, including the length of the incision, operating time for neck dissection, estimated blood loss during the operation, amount and duration of drainage, total hospitalization period, total number of lymph nodes retrieved, satisfaction scores based on the cosmetic results, perioperative local complications, shoulder syndrome, and follow-up information.The mean operation time in the endoscopically-assisted group (126.04 +/- A 12.67 min) was longer than that in the conventional group (75.67 +/- A 16.67 min). However, the mean length of the incision was 4.33 +/- A 0.76 cm in the endoscopically-assisted SND group, and the amount and duration of drainage, total hospital stay, postoperative shoulder pain score, and cosmetic outcomes were superior in the endoscopically-assisted SND group. Additionally, the retrieved lymph nodes and complications were comparable.Endoscopically-assisted SND via a small submandibular approach had a longer operation time than the conventional approach. However, endoscopically-assisted SND was feasible and reliable while providing minimal invasiveness and satisfactory appearance.

Molecular markers of angiogenesis and metastasis in lines of oral carcinoma after treatment with melatonin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)