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Eun Jee Chung, Hyung Keun Lee, Sun-Ah Jung, Seung Jae Lee, Hee Youn Chee, Yong Ho Sohn, Joon H. Lee; Transduction of PTEN Proteins Using the Tat Domain Modulates TGF-β1–Mediated Signaling Pathways and Transdifferentiation in Subconjunctival Fibroblasts. Invest. Ophthalmol. Vis. Sci. 2012;53(1):379-386. doi: 10.1167/iovs.11-8491.
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© ARVO (1962-2015); The Authors (2016-present)
This study investigated the effects of the tumor suppressor protein PTEN (phosphatase and tensin homolog) on transforming growth factor (TGF)-β1–mediated signaling pathways and the transdifferentiation of human subconjunctival fibroblasts (SCFs) after the transduction of this protein containing a transactivator of transcription (Tat) domain.
The Tat-PTEN expression vector was constructed to express the Tat domain of HIV-1 fused to PTEN. After transduction of the fusion protein and TGF-β1 stimulation, the dose-dependent effect of the transduced Tat-PTEN fusion protein on Akt phosphorylation and the stability of the Tat-PTEN fusion protein in SCF cells were evaluated by Western blot analysis. The effect of the Tat-PTEN fusion protein on the TGF-β1–stimulated expression of α-SMA and fibronectin was also evaluated by Western blot analysis and immunocytochemistry.
To increase the efficiency of enzyme activity and to successfully deliver this protein to cells, the authors used a PTEN fusion protein that contained the transduction domain of the Tat protein from HIV-1. By Western blot analysis, the transduced Tat-PTEN fusion protein was found to modulate TGF-β1 signaling in SCF cells and result in the suppression of Akt phosphorylation. Furthermore, the transduction of the Tat-PTEN fusion protein was found to suppress the TGF-β1–stimulated expression of α-SMA and fibronectin by Western blot analysis and immunocytochemical staining, and the effects of the transduced fusion protein could be controlled in a dose-dependent manner.
The Tat-PTEN fusion proteins were successfully transduced into the SCF cells and induced the suppression of transdifferentiation and fibrosis through the regulation of TGF-β–mediated signaling. The ability of the Tat-PTEN fusion protein to regulate cell survival could potentially be applied to protein therapy to counteract postoperative scarring in glaucoma surgery.
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