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Objective:To study the effects of activating protein kinase C (PKC)on oral squamous-cell cancer(OSCC) cell epithelial-mesenchymal transition(EMT).Methods:Plasmid pCMV6-AC-GFP-P120ctn was used to transfect HN12 cells to make P120-catenin (120ctn) overexpress and thereafter PKC activator PMA (phorbol 12-myristate 13-acetate) was added to the culture.Real-time fluorescent quantitative PCR and Western blot were adopted to test mRNA and protein expression of PKC,P120ctn,E-cadherin(E-cad),N-cadherin(N-cad) and Vimentin(Vim).Transwell cell invasion and cell migration assay were used to test the invasion and migration capacity before and after the activation.Results:When PKC was activated by PMA,the expression of P120ctn and E-cad were decreased,the cell morphology changed,nRNA and protein expression of mesenchymal marker protein N-cad and Vim increased significantly.Meanwhile,the migration and invasion capacity of the tumor cells increased significantly(P ＜ 0.05).Conclusion:In OSCC cells,PKC may be involved in promoting EMT and the metastasis and invasion by adjusting P120ctn/E-cad expression and cell adhesion.
ABSTRACT
<p><b>OBJECTIVE</b>The main goal is to investigate the role of P120-catenin (P120ctn) in cadherin switching, as well as migration and invasion, of oral squamous cell cancer (OSCC) cells.</p><p><b>METHODS</b>The plasmid pGFP-V-RS-P120ctn shRNA was used to transfect TSCCA cells and significantly reduce the expression of P120ctn in these cells. Real-time fluorescent quantitative polymerase chain reaction and Western blot were conducted to determine the mRNA and protein expression levels of P120ctn, E-cadherin (E-cad), and N-cadherin (N-cad). By contrast, the Transwell cell invasion and cell migration assay was used to determine the invasion and migration capacities before and after the transfection.</p><p><b>RESULTS</b>After the plasmid pGFP-V-RS-P120ctn shRNA was transfected into the TSCCA cells, we found that as the P120ctn expression significantly decreased, E-cad mRNA and protein expression decreased significantly. Moreover, N-cad mRNA and protein expression increased significantly (P<0.05). Lastly, the cell migration and invasion capacities were augmented significantly (P<0.05).</p><p><b>CONCLUSIONS</b>In OSCC cells, P120ctn may be involved in cadherin switching and promote metastasis and invasion.</p>
Subject(s)
Humans , Cadherins , Carcinoma, Squamous Cell , Catenins , Cell Line, Tumor , Cell Movement , Mouth Neoplasms , Neoplasm Invasiveness , Neoplasm Metastasis , TransfectionABSTRACT
Objective:To study the expression of IQ motif containing GTPase activating protein 1 (IQGAP1 )in oral squamous cell cancer(OSCC)tissue,and to explore the effects of IQGAP1 on cell proliferation and invasion as well as its underlying mechanism. Methods:Expression levels of IQGAP1 in tumor and adjacent normal tissues were examined by western blot and RT-PCR.OSCC cell line SCC-4 cells was transfected with the recombinant plasmid-pcDNA3.1 -IQGAP1 by lipofectamine,and then treated with an Akt in-hibitor.The phosphorylation of Akt,cell proliferation and invasion were detected by western blot,MTT assay and Transwell invasion as-say respectively.Results:Protein and mRNA expression levels of IQGAP1 were higher in cancer tissue than in adjacent normal tissue (P <0.05).Transfection of pcDNA3.1 -IQGAP1 increased IQGAP1 expression,enhanced the capability of cell proliferation and inva-sion (P <0.05),increased p Akt level in the cells.Preconditioning with an Akt inhibitor reduced p Akt level.Furthermore,silencing Akt pathway blocked the increase of cell proliferation and invasion induced by IQGAP1 overexpression(P <0.05).Conclusion:IQ-GAP1 overexprission can mediate the ability of proliferation and invasion of OSCC cells by regulating the activation of Akt pathway.
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Das diversas neoplasias denominadas câncer de boca, mais de 95% tem o diagnóstico de carcinoma epidermóide (CE). O carcinoma epidermóide apresenta etiologia e patogênese complexas e não totalmente compreendidas determinando altos índices de morbidade e mortalidade. Por isso crescentes esforços têm sido empregados para a melhor compreensão dos eventos celulares, da susceptibilidade genética e dos fatores de risco que atuando em conjunto dão origem e atuam na progressão do carcinoma epidermóide de boca. Buscando novas facetas da carcinogênese oral e baseando-se no papel pró-carcinogênico que os hormônios esteróides desempenham nas neoplasias dos órgãos reprodutivos, alguns estudos passaram a avaliar o papel desses hormônios na patogênese e progressão do CE. O propósito deste trabalho foi avaliar a presença dos receptores de androgênio, estrogênio (alfa e beta) e da enzima aromatase em carcinomas epidermóides de boca (CE), através das técnicas de imuno-histoquímica, western-blotting e imunofluorescência, e comparar, com base no gênero (masculino e feminino), os resultados encontrados. Da amostra de CE utilizada, 30 eram pertencentes a pacientes do gênero feminino e 30 do masculino...
Oral squamous cell carcinoma (OSCC) is the main type of oral cancer presenting complex and not completely understood pathogenesis. OSCC implies quite significant mortality and morbidity rates and in spite of the vast amount of research and the advances in the field of oncology and surgery, the overall survival remains largely unchanged. Therefore, the identification of new pathways involved in the mechanisms of carcinogenesis can bring knowledge to the control and advent of new treatments. The role of androgens and estrogens in several endocrine-related malignancies is well known. The successful use of these hormones antagonists in the treatment of these neoplasms has prompted researches to investigate the presence of hormones receptors in other tissues and tumor types. The aim of this study was to asses through immunohistochemistry, western blot and immunoflorescence assays the expression, genderrelated, of androgen, estrogen alpha and beta receptors and aromatase enzyme in cases of OSSC. A total of 60 cases of OSCC (30 from males and 30 from females) were retrieved and submitted to immunohistochemical assay. Two OSCC cell lines were used for western blot and immunoflurescence techniques...
Subject(s)
Humans , Male , Female , Aromatase , Carcinoma, Squamous Cell/diagnosis , Pathology, Oral/methodsABSTRACT
Objective:To investigate the expression of tumor necrosis factor-?(TNF-?)in the tissues of oral squamous cell carcinoma(OSCC)and its clinical significance. Methods:The expression of TNF-? was assessed by immunohistochemical method of SABC in paraffin tissue sections of 41 cases of OSCC and 10 of normal oral epithelia.Results:The expression of TNF-? in OSCC was higher than that in oral normal mucous tissue(P