April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Mitomycin C Retards Corneal Fibroblast Cell Growth by Endogenous Fibulin-1 Expression
Author Affiliations & Notes
  • Shu-Wen Chang
    Dept of Ophthal, Far Eastern Memorial Hospital Library, Ban-Chiao, Taipei, Taiwan
    Dept of Ophthal, National Taiwan University Hospital, Taipei, Taiwan
  • Tsan-Chi Chen
    Dept of Ophthal, Far Eastern Memorial Hospital Library, Ban-Chiao, Taipei, Taiwan
  • Han-Fang Teng
    Dept of Ophthal, Far Eastern Memorial Hospital Library, Ban-Chiao, Taipei, Taiwan
  • Chih-Chieh Lee
    Dept of Ophthal, Far Eastern Memorial Hospital Library, Ban-Chiao, Taipei, Taiwan
  • Pei-Hsiu Chiang
    Dept of Ophthal, Far Eastern Memorial Hospital Library, Ban-Chiao, Taipei, Taiwan
  • Footnotes
    Commercial Relationships Shu-Wen Chang, None; Tsan-Chi Chen, None; Han-Fang Teng, None; Chih-Chieh Lee, None; Pei-Hsiu Chiang, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 5149. doi:
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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)

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Abstract
 
Purpose
 

To investigate the role of fibulin-1 (FBLN1) in mitomycin C (MMC)-modulated cell growth in human corneal stromal cells (HCFs).

 
Methods
 

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.

 
Results
 

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.

 
Conclusions
 

MMC-related diminution of the intracellular FBLN1 expression might suppress the cell proliferation in HCFs.

 
 
Figure 1. Expression of endogenous FBLN1 in HCFs. (A) In normal HCFs, cell lysates were harvested at the indicated time points by immunoblotting assay. We found FBLN1 expression increased up to 72 h in cultivation in normal HCFs. (B) Quantitative data were expressed as the mean ± S.E.M. of three individual experiments. Differences in the relative change of FBLN1 were analyzed by one-way ANOVA and compared with the groups at 0 h (**, P < 0.01; ***, P < 0.001).
 
Figure 1. Expression of endogenous FBLN1 in HCFs. (A) In normal HCFs, cell lysates were harvested at the indicated time points by immunoblotting assay. We found FBLN1 expression increased up to 72 h in cultivation in normal HCFs. (B) Quantitative data were expressed as the mean ± S.E.M. of three individual experiments. Differences in the relative change of FBLN1 were analyzed by one-way ANOVA and compared with the groups at 0 h (**, P < 0.01; ***, P < 0.001).
 
Keywords: 449 cell survival • 480 cornea: basic science • 654 proliferation  
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