Fol. Biol. 2008, 54, 46-52
Quantitative Analysis of Transforming Growth Factor β Isoforms mRNA in the Human Corneal Epithelium
TGF-β is an important mediator of cell growth, differentiation, and proliferation and plays a significant role in both normal and pathological corneal tissue. However, the quantitative relations between TGF-β1, -β2 and -β3 isoforms in human cornea still remain unclear. Therefore, the aim of this study was to determine the gene expression profile of TGF-βs in order to evaluate quantitative relations between the examined transcripts in human corneal epithelium. Transcriptional activity of TGF-β1, 2, 3, GAPDH and β-actin genes was estimated on the basis of mRNA copy number per 1 μg of total RNA using the real-time QRT-PCR technique with the SYBR Green I chemistry. Specificity of RT-PCR reaction was confirmed by determination of the characteristic melting temperature for each amplimer. Additionally, the RT-PCR products were separated on 6% polyacrylamide gels and visualized with silver salts. Expression of all TGF-β genes for the corneal epithelium was determined. Comparable analysis of mRNA copies/1 μg of total RNA for each TGF-β isoform showed that: TGF-β1 > TGF-β2; TGF-β3 > TGF-β2; TGF-β1 = TGF-β3 (ANOVA test P < 0.0001; post-hoc Tukey’s test: TGF-β1 and TGF-β2, P = 0.0306; TGF-β3 and TGF-β2, P = 0.0045; TGF-β1 and TGF-β3 NS). We found different expression of the TGF-β1, -2 and -3 isoforms in the human corneal epithelium. Such differential expression of TGF-βs suggests that each of them may play a specific role in corneal tissue.
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