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J. Yun, J.-E. Im, C. Kee; Effect of Heat Shock Protein 72 Expression Against Etoposide-Induced Apoptotic Cell Death of Rat Retinal Ganglion Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2791.
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To assess whether the expression of heat shock protein 72 (HSP72) protects rat retinal ganglion cells from apoptotic cell death.
Rat retinal ganglion cell line (RGC-5) was transduced with replication-deficient recombinant adenovirus expressing human HSP72 (approved gene symbol HSPA1A) at an MOI of 10 ~ 100 pfu, washed once, and then exposed to 5 ~ 50 uM etoposide. After 24 hours, HSP72 expression was analyzed by Western blot analysis and immunocytochemistry. The effect of HSP72 expression on the cell survival rate was evaluated by MTT assay and confirmed by microscopic analysis.
Western blot analysis and immunocytochemistry clearly showed adenovirus-mediated HSP72 expression in RGC-5. The treatment of etoposide, an inhibitor of topoisomerase II, resulted in apoptosis in approximately 58 ± 8% of untransduced cells. However, the apoptotic cell death was significantly reduced by 45 ± 6% ~ 36 ± 11% (p <0.01) in cells expressing HSP72 with the degree of reduction in cell death correlated with the amount of HSP72 protein expressed, demonstrating that HSP72 inhibited etoposide-induced apoptotic cell death.
Expression of HSP72 alone can protect retinal ganglion cells against apoptotic cell death, suggesting that its expression potentially paves the road for novel therapeutic approach to glaucoma.
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