Regulatory mechanism of human connective tissue growth factor (CTGF/Hcs24) gene expression in a human chondrocytic cell line, HCS-2/8
| Autor: | Takuo Kuboki, Tohru Nakanishi, Takako Hattori, Seiji Kondo, Satoshi Kubota, Hirofumi Yatani, Masaharu Takigawa, Tsuyoshi Shimo, Takanori Eguchi |
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| Rok vydání: | 2001 |
| Předmět: |
Untranslated region
Transcription Genetic Response element Response Elements Transfection Biochemistry Cell Line Immediate-Early Proteins Chondrocytes Transcription (biology) Transforming Growth Factor beta Gene expression Humans Point Mutation Luciferase RNA Messenger Growth Substances Promoter Regions Genetic Molecular Biology 3' Untranslated Regions Sequence Deletion Regulation of gene expression integumentary system Base Sequence Chemistry Reverse Transcriptase Polymerase Chain Reaction Connective Tissue Growth Factor General Medicine Cell biology CTGF Repressor Proteins Gene Expression Regulation Intercellular Signaling Peptides and Proteins Electrophoresis Polyacrylamide Gel HeLa Cells |
| Zdroj: | Scopus-Elsevier |
| ISSN: | 0021-924X |
| Popis: | CTGF/Hcs24 is a multi-functional growth factor that potentiates either the growth or differentiation of mesenchymal cells, according to the biological conditions. Among various functional aspects of CTGF/Hcs24, it is especially notable that CTGF/Hcs24 may promote endochondral ossification in growth cartilage through all stages, and it is highly expressed in a human chondrosarcoma-derived chondrocytic cell line (HCS-2/8). In this study, to clarify the regulatory mechanism of CTGF/Hcs24 gene expression in chondrocytes, we analyzed the transcriptional activity of the CTGF/Hcs24 promoter and the effect of the CTGF/Hcs24 3'-untranslated region (3'-UTR) on gene expression in HCS-2/8 by means of an established DNA transfection and luciferase reporter gene assay system. As a result, the luciferase activity of the CTGF/Hcs24 promoter was found to be remarkably high in HCS-2/8. The 3'-UTR of the CTGF/Hcs24 gene strongly repressed the luciferase activity in HCS-2/8, when it was linked to the downstream of the luciferase reporter gene, suggesting its functionality also in chondrocytic cells. Deletion analysis of the CTGF/Hcs24 promoter clarified a major segment responsible for the enhanced CTGF/Hcs24 promoter activity in HCS-2/8. The TGF-beta response element in the DNA segment was active in HCS-2/8, and point mutations in the element moderately decreased the highly maintained promoter activity with total loss of TGF-beta responsiveness. These results indicate that the strong expression of the CTGF/Hcs24 gene in HCS-2/8 was mainly caused by high transcriptional activity of the CTGF/Hcs24 promoter, and that the TGF-beta response element is one of the critical elements that support the high transcription activity. |
| Databáze: | OpenAIRE |
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