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Shu-Wen Chang, Tsan-Chi Chen, Han-Fang Teng, Chih-Chieh Lee, Pei-Hsiu Chiang; Mitomycin C Retards Corneal Fibroblast Cell Growth by Endogenous Fibulin-1 Expression. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5149.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the role of fibulin-1 (FBLN1) in mitomycin C (MMC)-modulated cell growth in human corneal stromal cells (HCFs).
HCFs were treated with MMC at 0.2 mg/ml and cultivated in DMEM with 10% fetal bovine serum. Transcriptional and protein expressions of FBLN1 were analyzed by real-time PCR and immunoblotting. The lentivirus-based pseudovirion-based infection system was used for overexpressing and silencing FBLN1 expression in HCFs. Cell adhesion capability of normal, FBLN1-expressed, or FBLN1-silenced HCFs was monitored by real-time cultured cell monitoring system. Cell growth was determined daily by WST-1 cell proliferation assay.
MMC dose-dependently inhibited cell proliferation of HCFs. FBLN1 expression increased with cultivation time in normal HCFs (Figure 1). MMC significantly suppressed FBLN1 at transcriptional level and protein expression in a dose-dependent manner. The time lapse images revealed that increased expression of FBLN1 enhanced cell adhesion and promoted cell growth. On the contrary, silence of FBLN1 retarded cell adhesion and inhibited cell growth.
MMC-related diminution of the intracellular FBLN1 expression might suppress the cell proliferation in HCFs.
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