Purpose: To evaluate the role of miR-200b expression in the proliferation of human Tenon's capsule fibroblasts (HTFs) induced by transforming growth factor-beta 1 (TGF-β1).
Methods: Human Tenon's capsule fibroblasts were treated with various doses of TGF-β1 for 24 hours. Cell proliferation was quantified by the cell counting kit-8 (cck-8) assay, cell cycle analysis, 5-ethynyl-2-deoxyuridine (EdU) assay, and analysis of cyclin E, cyclin D1, and proliferating cell nuclear antigen (PCNA) expression. MicroRNA-200b (miR-200b) was detected by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), and its potential target genes were validated by the luciferase assay and Western blot analysis. The effect of miR-200b on the proliferation of HTFs was analyzed using both miR200b-mimic and inhibitor-transfected HTFs and confirmed in p27/kip1 and RND3 (the target of miR-200b) knockdown cells.
Results: The proliferation of the TGF-β1-treated HTFs increased significantly in a dose- and time-dependent manner. Treatment with 5 ng/mL TGF-β1 caused an upregulation of miR-200b. The luciferase assay identified p27/kip1 and RND3 as target genes for miRNA-200b, which was confirmed by the expression of p27/kip1 and RND3 and their downstream products (cyclinE and cyclinD1) in the TGF-β1-treated cells. Transforming growth factor-β1 and miR-200b mimics enhanced the proliferation of HTFs; suppressed the expression of p27/kip1 and RND3; and subsequently stimulated the expression of cyclinE, cyclinD1, and PCNA. The miR-200b inhibitor attenuated the effects of TGF-β1 on HTFs. Furthermore, knockdown of p27/kip1 and RND3 resulted in an increase in cell proliferation and expression of the proliferation-related genes.
Conclusions: MicroRNA-200b acts as a stimulant for the proliferation of HTFs by targeting p27/kip1 and RND3.
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http://dx.doi.org/10.1167/iovs.13-13422 | DOI Listing |
Biomed Pharmacother
October 2018
Digestive Department; Tongchuan traditional Chinese medicine hospital, Shaanxi, People's Republic of China. Electronic address:
Exosome are emerging mediators of intercellular communication. Cancer-secreted exosome has an effect on the exosome donor cells and support cancer growth and metastasis. Here, we examine the TGF-β1, a multifunctional cytokine involved in the regulation of cellular signaling pathways in human cancers, significantly contributes to upregulate miR-200b in exosome from colorectal cancer cell lines.
View Article and Find Full Text PDFInvest Ophthalmol Vis Sci
April 2014
Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China.
Purpose: To evaluate the role of miR-200b expression in the proliferation of human Tenon's capsule fibroblasts (HTFs) induced by transforming growth factor-beta 1 (TGF-β1).
Methods: Human Tenon's capsule fibroblasts were treated with various doses of TGF-β1 for 24 hours. Cell proliferation was quantified by the cell counting kit-8 (cck-8) assay, cell cycle analysis, 5-ethynyl-2-deoxyuridine (EdU) assay, and analysis of cyclin E, cyclin D1, and proliferating cell nuclear antigen (PCNA) expression.
J Cell Physiol
June 2014
Department of Physiology, Nanjing Medical University, Nanjing, People's Republic of China.
Colorectal cancer (CRC) remains the most common malignancy worldwide. TGF-β1 is often overexpressed in late stages of colorectal carcinogenesis and promotes tumour growth and metastasis. Several reports have verified that the loss of functional TGFBRII expression contributed to escape the tumour suppressor activity of TGF-β1 and that the epithelial-to-mesenchymal transition (EMT) responded to TGF-β1 involved in tumour invasion and metastasis.
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