HSV-1 infection suppresses TGF-beta1 and SMAD3 expression in human corneal epithelial cells.
OBJECTIVE
The present study was undertaken to investigate whether transforming growth factor-beta (TGF-beta) isoforms (TGF-beta1, TGF-beta2, and TGF-beta3) and SMADs (SMAD2 and SMAD3) are involved in herpes simplex virus type 1 (HSV-1) corneal infection.


METHODS
Human corneal epithelial cells (HCE) were infected with HSV-1 at a multiplicity of infection of 5. Cell morphological changes were observed under phase-contrast microscopy. Levels of mRNA for TGF-beta isoforms 1, 2, and 3 as well as for SMAD2 and SMAD3 were measured by reverse transcription polymerase chain reaction (RT-PCR) at 0 h, 4 h, 8 h, 12 h, and 24 h after infection. Protein expression of TGF-beta1, TGF-beta2, SMAD3, and phospho-SMAD3 were analyzed by indirect immunofluorescence at 0 h, 12 h, and 24 h post-infection (p.i.) in HCE cells. Protein expression of TGF-beta1 was also evaluated by ELISA.


RESULTS
Following HSV-1 infection, a cytopathic effect in HCE cells was observed at 8 h p.i. and became significant at 24 h p.i. Compared with normal cells, the mRNA levels of TGF-beta1 in HSV-1 infected HCE cells decreased significantly at 8 h, 12 h, and 24 h p.i. (p<0.01), and the expression of SMAD3 was also dramatically decreased 12 h and 24 h p.i. (p<0.01). No noticeable changes were found as a result of infection with respect to levels of TGF-beta2, TGF-beta3, and SMAD2 in HCE cells. Protein expression of TGF-beta1, SMAD3, and phospho-SMAD3 decreased in infected cells at 12 h and 24 h p.i. compared with normal cells, but TGF-beta2 had no change.


CONCLUSIONS
TGF-beta1 and SMAD3 may be involved in the pathology of corneal diseases associated with HSV-1 infection.
Nie
Yuhong
Y
Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, PR China.
Cui
Dongmei
D
Pan
Zhujuan
Z
Deng
Jiangyun
J
Huang
Qiang
Q
Wu
Kaili
K
D002460:Cell Line; D048430:Cell Shape; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism* / Q000821:virology*; D019573:Epithelium, Corneal / Q000166:cytology* / Q000821:virology*; D005786:Gene Expression Regulation; D018259:Herpesvirus 1, Human / Q000502:physiology*; D006801:Humans; D010750:Phosphoproteins / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*; D053781:Transforming Growth Factor beta2 / Q000235:genetics / Q000378:metabolism; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism
D010750:Phosphoproteins; D020033:Protein Isoforms; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D053773:Transforming Growth Factor beta1; D053781:Transforming Growth Factor beta2; D053782:Transforming Growth Factor beta3
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