コラーゲンスポンジを用いた三次元培養におけるヒト顎下腺由来腺癌細胞株の遺伝子発現

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Expressions of genes and proteins of human salivary gland cells (HSG cells) were investigated on three-dimensional culture comprised in collagen sponge in order to understand the response of simple epithelial cells at the molecular level. HSG cells were seeded in collagen sponges (COL-S)(three-dimensional culture) with an agitated seeding method, and cultured for various times. As a control, HSG cells were cultured on a polystyrene dish (PS) or a collagen coated dish (COL-C)(monolayer culture). The morphological features indicate that HSG cells cultured with COL-S presented a globular shape compared with monolayer culture such as with PS or COL-C. We focused on cytokeratins (CK) that are used as the marker proteins for epithelia to characterize the cultured cells. In HSG cells, CK7, CK8, CK17 and CK18 are expressed, but CK13 which is expressed specifically in mucosal epithelia is not expressed. The expression levels of CK genes were investigated by RT-PCR. The results were that the level of CK8 and CK18 were downregulated. The level of CK13 was upregulated. On the other hand, no alternation of the expression levels of CK7 and CK17 was observed. Western Blotting was carried out to investigate the expression of protein. The bands of CK8 and CK18 were detected. The expression level of CK8 and CK18 protein was reduced as well as the mRNA level. CK13 protein was not detected in this way. The expression level of the Nm23-H1 gene, which is one of tumor suppressor genes and concerned with ERK/MAPK signal transduction, was downregulated. These results suggest that HSG cells were altered by three-dimensional culture for different expression patterns of CKs which were not contained in the simple epithelium.
岩手医科大学歯学会の論文
2005-12-20

コラーゲンスポンジを用いた三次元培養におけるヒト顎下腺由来腺癌細胞株の遺伝子発現

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