This study suggests the participation of pAkt, NF-κB and cyclin D1 in the invasion process of head and neck squamous cell carcinoma. Moreover, cytoplasmic pAkt network-like structure was probably related to cytoskeleton changes presented during invasion.
Vimentin is a cytoeskeletal intermediate filament protein commonly observed in mesenchymal cells; however, it can also be found in malignant epithelial cells. It is demonstrated in several carcinomas, such as those of the cervix, breast and bladder, in which it is widely used as a marker of the epithelial to mesenchymal transition that takes place during embryogenesis and metastasis. Vimentin is associated with tumors that show a high degree of invasiveness, being detected in invasion front cells. Its expression seems to be influenced by the tumor microenvironment. The aim of this study was to evaluate vimentin expression in head and neck squamous cell carcinoma (HNSCC) cell lines, and to investigate the contribution of the microenvironment to its expression. HNSCC cell lines (HN6, HN30 and HN31) and an immortalized nontumorigenic cell line (HaCaT) were submitted to a three-dimensional assay with Matrigel. Cytoplasmatic staining of the HN6 cell line cultured without Matrigel and of the HN30 and HN31 cell lines cultured with Matrigel was demonstrated through immunohistochemistry. Western Blotting revealed a significant decrease in vimentin expression for the HN6 cell line and a significant increase for the HN30 and HN31 cell lines cultured with Matrigel. The results suggest that vimentin can be expressed in HNSCC cells and its presence is influenced by the microenvironment of a tumor.
Within the results encountered, the present panel should be of great assistance in the diagnosis of oral IMTs.
Head and neck squamous cell carcinoma (HNSCC) represents more than 90% of all head and neck malignancies, causing more deaths than any other oral disease. Proteins related to cancer growth, invasion and metastasis are in evidence due to their involvement in carcinogens, such as vimentin. This protein is observed in mesenquimal cells, however, it is considered a common finding in cervix, breast and bladder tumours. Thus its presencence in epithelial neoplasic cells contributes to epithelial-mesenchymal transition associated with tumorigenesis and tumor progression. The aim of this study was to analyse through Western Blot, Immunohistochemistry and Immunofluorescence methods, the expression of Vimentin in three different HNSCC cell lines and HaCat cell line (immortalized keratinocytes) submitted to a 3D assay into Matrigel®. The control group was represented by the same cell lines, without any treatment. Results showed that Vimentin had citoplasmatic staining in some cell of lines studied, except for HaCat cells, with evident decrease in its expression when submitted to cultive into Matrigel®. These findings were confirmed by Western Blot. Taking these results together, we conclude that in squamous cell carcinoma, the Vimentin is related to the process of tumour invasion and metastasis. This fact was showed by the reduction of its expression after treatment with Matrigel®. Therefore, the expression of Vimentin in different cell lines of HNSCC may vary according to the stimulus and, fundamentally, the localization of the tumor and the individual characteristics of neoplasic cells.
RESUMO O Celecoxib, antiinflamatório não esteroidal, inibidor seletivo da COX-2, tem se mostrado um importante agente anticarcinogênico, mas o seu papel no carcinoma epidermóide de boca (CEB) não é totalmente compreendido. Sabe-se que diversas alterações genéticas estão associadas à patogênese do CEB, a neoplasia maligna mais comum de cabeça e pescoço. Algumas dessas alterações comprometem proteínas pertencentes à via de sinalização do Akt, envolvida em diferentes fenômenos celulares. É sabido que Akt pode ativar o fator de transcrição NF-kB, o qual tem importante participação na fisiologia normal e no câncer. A proteína COX-2, descrita inicialmente em processos inflamatórios, está associada com a oncogênese e recentemente tem sido associada com a via de sinalização do Akt e com o NF-kB. Portanto, o objetivo deste estudo foi analisar o efeito do Celecoxib sobre linhagens celulares de carcinoma epidermóide de boca e verificar a localização intracelular e a expressão das proteínas pAkt, NF-kB e COX-2 em linhagens celulares de carcinoma epiermóide de boca após o tratamento com o Celecoxib.Através da técnica de imunofluorescência, foram analisados os padrões de expressão das proteínas pAkt, NFkB e COX-2 em quatro linhagens celulares de carcinoma epidermóide bucal submetidas ao tratamento com Celecoxib, cuja a dose e o tempo foram obtidos a partir de ensaios de viabilidade celular. Também se realizou ensaio de apoptose celular. Como controle utilizou-se células não tratadas com o medicamento. O Celecoxib na dose de 30 µM por 24 horas causa apoptose.Na técnica de western blot, somente a linhagem SCC15 apresentou uma diminuição significativa para a COX-2. Entretanto, para p-Akt e NF-kB nenhuma alteração na expressão foi observada entre os grupos controle e tratado.Na imunofluorescência, houve alteração no padrão de expressão das proteínas pAkt, NF-kB e COX-2, quando se comparou os grupos contrele e tratado. Portanto, o Celecoxib pode ser um eficaz agente terapêutico, uma vez que demonstrou grande eficácia na inibição da proliferação celular de linhagens celulares de CEB. Palavras-chave: Carcinoma epidermóide. Celecoxib. Via de sinalização p-Akt/NF-kB. COX-2. apoptose. Vechio AMCD. Apoptotic effect of Celecoxib in cell lines derived from oral squamous cell carcinoma [thesis].
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