RESUMEN
The extent of lymph node metastasis is a major determinant in the prognosis of oral squamous cell carcinoma (OSCC). Abnormalities of cell adhesion molecules are known to play an important role in invasion and metastasis of cancer cells through the loss of cell-to-cell adhesion. In this study, we isolated highly invasive clones from an OSCC cell line established from a lymph node metastasis by using an in vitro invasion assay method and compared the abnormalities of cell adhesion molecule E-cadherin and beta-catenin in these cells. The isolated, highly invasive clones showed significant invasive capacity and reduction of E-cadherin and membranous beta-catenin protein in comparison with parent cells. We found that reduced expression of E-cadherin was due to methylation of its promoter region. In fact, most invasive and metastatic area of OSCCs showed reduced expression and methylation of E-cadherin. Moreover, we found that reduced expression of membranous beta-catenin was due to its protein degradation. Reduced expression of membranous beta-catenin was also found frequently in invasive and metastatic areas of OSCCs. In summary, invasion and metastasis of OSCC cells require methylation of E-cadherin and/or degradation of membranous beta-catenin. In addition, we suggest that the method of isolation of highly invasive clones may be useful for studies aimed at discovering novel genes involved in invasion and metastasis.
Asunto(s)
Cadherinas/genética , Neoplasias Gingivales/genética , Neoplasias Gingivales/patología , Cadherinas/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Proteínas del Citoesqueleto/metabolismo , Metilación de ADN , Silenciador del Gen , Células HeLa , Humanos , Neoplasias de la Boca/genética , Neoplasias de la Boca/patología , Invasividad Neoplásica , Metástasis de la Neoplasia , Regiones Promotoras Genéticas/genética , ARN Interferente Pequeño/genética , Transactivadores/metabolismo , beta CateninaRESUMEN
OBJECTIVES: CD44 is one of the cell surface molecules that play an important role in cancer metastasis. In oral squamous cell carcinoma (OSCC), the downregulation of CD44v9 has been shown to be associated with tumor metastasis. We found that treatment with an anti-CD44v9 antibody enhanced the invasive potential of OSCC cell lines. Based on previous studies and our results, reduced expression of CD44v9 may be correlated with an increased invasive potential, i.e. the overexpression of CD44v9 may inhibit the invasive activity of OSCC cells. METHODS: To study this correlation, we transfected the CD44v9 gene into HSC-4 cells with low CD44v9 expression and examined their invasive potential using the cell culture invasion assay and a three-dimensional culture invasion assay. RESULTS: Overexpression of CD44v9 resulted in a downregulation of the invasive potential of HSC-4 cells. Moreover, CD44v9-transfected cells did not invade reorganized stroma, while parent HSC-4 cells exhibited diffuse invasion into reorganized stroma by the three-dimensional culture invasion assay. CONCLUSIONS: Overall, these findings suggest that the inhibition of the invasive potential by upregulation of CD44v9 expression may be due to enhanced cell-cell adhesion. In our opinion, the upregulation of CD44v9 may be a target for future cancer treatment.
Asunto(s)
Carcinoma de Células Escamosas/patología , Regulación Neoplásica de la Expresión Génica , Receptores de Hialuranos/genética , Neoplasias de la Boca/patología , Western Blotting , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Adhesión Celular , Línea Celular Tumoral , Humanos , Receptores de Hialuranos/metabolismo , Inmunohistoquímica , Neoplasias de la Boca/genética , Neoplasias de la Boca/metabolismo , Invasividad Neoplásica , Transfección , Regulación hacia ArribaRESUMEN
This study investigated the recruitment of polymorphonuclear leukocytes (PMNs) and the immunolocalization of CXC chemokines, including macrophage inflammatory protein-2 (MIP-2) and cytokine-induced neutrophil chemoattractant-2 (CINC-2) in rat periodontal tissue after topical application of lipopolysaccharide (LPS; 5 mg/ml) from Escherichia coli into the rat molar gingival sulcus. In normal periodontal tissues, a small number of MIP-2- and CINC-2-positive cells were seen in junctional epithelium (JE), especially in its coronal half. After topical application of LPS, a prominent increase of MIP-2- and CINC-2-positive JE cells was observed. Almost all JE cells strongly expressed them at day 1 and day 2, and then the number of chemokine-positive cells returned to normal at day 7. Corresponding to these chemokine expressions, LPS application induced a significant increase in the number of PMNs in the sub-JE area from 1 h to 2 days and a significant increase in JE area from 3 h to 5 days, indicating a dynamic flow of PMNs from the sub-JE area into JE. These findings indicated that JE cells produced MIP-2 and CINC-2 in response to LPS stimulation and suggested that MIP-2 and CINC-2 may be responsible for PMN migration toward the periodontal pathogen and may play an important role in the initiation of inflammation and subsequent periodontal tissue destruction.