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J Wang, H Xiang, Y Mao, DW Li; Prevention Of Cellular Senescence And Cell Differentiation By hTERT Is Associated With Modulated MAP Kinase Activity . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2353.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Our previous studies have revealed that the human telomerase catalytic subunit (hTERT), when transfected into various lens epithelial cells, display different functions such as retardation of induced cell death, modulation of gene expression and acceleration of cell growth. To further study the function of hTERT in lens system, We have established bovine and human lens epithelial cell lines expressing hTERT. Using hTERT- and vector-transfected bovine lens epithelial cell line, we have demonstrated some other functions of hTERT. Methods: Primary cultures of bovine and human lens epithelial cells were transfected with an exogenous hTERT gene which is under regulation of a CMV promoter. Individual stable expression clones were screened with G418. Established clones of hTERT- and vector-transfected cells were used to study the functions of hTERT. Cell growth rate was monitored after a certain period of passages. Cellular senescence was determined by SA-beta-gal staining. Cell differentiation was detected by examination of beta-crystallin gene expression. The involvement of MAP kinase pathway in mediating hTERT function was determined with specific inhibitors and Western blot analysis. Results: RT-PCR was used to verify expression of the exogenous hTERT gene in different clones. TRAP assay and immunoprecipitation-linked RT-PCR confirmed that hTERT is functionally compatible with bovine telomerase template RNA. After about 20 PDLs, the vector-transfected bovine lens epithelial cells ran into cellular senescence or differentiated status as indicated by positive SA-beta-gal staining or expression of beta-crystallin gene. Significantly higher level of MAP kinase activity was also observed in vector-transfected cells. However, the hTERT-transfected cells display normal growth pattern. Conclusions: The hTERT-regulated MAP kinase activity is associated with prevention of cellular senescence and cell differentiation in bovine lens epithelial cells. Supported by EY 11372, NJ Foundation and UMDNJ/GSBS. None.
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