No abstract

The role and prognostic value of tetraspanins (TSPANs) in vulvar squamous cell carcinoma (VSCC) remain poorly understood. We sought to primarily determine, at both the molecular and tissue level, the expression profile of the TSPANs CD9, CD63, CD81, and CD82 in archived VSCC samples (n = 117) and further investigate their clinical relevance as prognostic markers. Our studies led us to identify CD63 as the most highly expressed TSPAN, at the gene and protein levels. Multicomparison studies also revealed that the expression of CD9 was associated with tumor size, whereas CD63 upregulation was associated with histological diagnosis and vascular invasion. Moreover, low expression of CD81 and CD82 was associated with worse prognosis. To determine the role of TSPANs in VSCC at the cellular level, we assessed the mRNA levels of CD63 and CD82 in established metastatic (SW962) and non-metastatic (SW954) VSCC human cell lines. CD82 was found to be downregulated in SW962 cells, thus supporting its metastasis suppressor role. However, CD63 was significantly upregulated in both cell lines. Silencing of CD63 by siRNA led to a significant decrease in proliferation of both SW954 and SW962. Furthermore, in SW962 particularly, CD63-siRNA also remarkably inhibited cell migration. Altogether, our data suggest that the differential expression of TSPANs represents an important feature for prognosis of VSCC patients and indicates that CD63 and CD82 are likely potential therapeutic targets in VSCC.

, minha orientadora, agradeço pela principalmente pela confiança que depositou em mim desde o começo dessa jornada. Agradeço por ter sido muitas vezes compreensiva, pelos ensinamentos científicos (prático e teórico) e pela paciência. Obrigada por entregar em minhas mãos uma missão tão bonita quanto essa, pesquisar métodos para tornar melhor a vida de muitas mulheres. Agradeço aos Professores Dr. Edmund Chada Baracat e Dr. Gustavo Arantes Rosa Maciel por terem aberto as portas do laboratório de Ginecologia-LIM 58 e por terem me concedido a oportunidade de fazer parte desta grande família. Agradeço por todo suporte e incentivo em continuar a caminhada da pósgraduação. Ao Prof. Dr. Rafael Malagoli Rocha, pela gentileza em ceder as células de carcinoma de vulva para o desenvolvimento deste trabalho. Ao Dr. Antônio Augusto Rocha, pela conduta e suporte científico com os protocolos de extração proteica e transfecção com RNAi, e por todas as vezes que, com muita paciência, respondeu a todos os meus questionamentos. À Dra. Mara Hoshida, e aos funcionários do Laboratório de Fisiologia Obstétrica-LIM57, Salete, Cristiane e Isaías, pelo consentimento na utilização do laboratório de cultura celular, enquanto nosso laboratório estava em reforma. Agradeço ao grande amigo, padrinho de casamento e padrinho científico, Leonardo Tomiatti da Costa, pelos anos de amizade, respeito, carinho e consideração que teve por mim quando me indicou para trabalhar no LIM58. Pela

Background: Uterine leiomyoma (ULM) and leiomyosarcoma (ULMS) are smooth muscle tumors with distinct clinical and biological behavior. They can be a cause of infertility and even death. Little is known about the factors that can to influence these tumors behavior and biology. Sonic Hedgehog (SHH) pathway components were previously implicated in the ULMS malignancy risk; however its activation mechanism is unknown. In this study, we investigated the gene expression profile of Sonic Hedgehog Signaling and pathway upstream genes in myometrium (MM), ULM and LMS cell lines. Methods: Total RNA obtained from cell lines were submitted to cDNA synthesis and quantitative real time PCR (qRT-PCR), using 7500 System (Life Technologies, USA). Gene expression of SHH, PTCH1, SMO, SUFU, GLI 1-3, CCND1, BCL-2 and BMP4 were assessed. Results: Gene expression data showed an upregulation of SMO, SUFU, GLI1, BCL-2 and BMP4 in LMS cells when compared to MM. GLI1 (RQ=11) and BCL-2 (RQ=20) genes had higher expression. In the other side, SHH, PTCH1, GLI2 and GLI3 showed upregulation in MM cells. Conclusion: Our results showed that the Sonic hedgehog pathway is activated in LMS cells but by SHH independent form (non-canonical pathway). This could be explained by the downexpression of SHH ligand and the PTCH1 receptor, and by the upexpression of SMO, GLI1 and pathway target genes as BCL-2 and BMP4 in uterine LMS cells compared to LM and MM. These results suggest a new therapeutic perspective for LMS by target drugs to inhibit SMO and GLI1 molecules. Citation Format: Natalia Garcia, Bianca C. Oliveira, Kelly P. Ferreira, Edmund C. Baracat, Katia C. Carvalho. Non canonical activation of Sonic Hedgehog pathway in uterine leiomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2436. doi:10.1158/1538-7445.AM2017-2436