Induced Expression Of Pcdna3.1-egr1-egfp-omi/htra2 And Its Effect On Apoptosis Of Human Choroidal Melanoma Ocm-1 Cells

Objective:To observe the changes of apoptosis of human choroidal melanoma cells (OCM-1) transfected with pcDNA3.1-Egr1-EGFP-Omi/HtrA2 plasmid after induced by hydrogen peroxide, to explore the induced and the kill characteristics of Oxygen free radicals of pcDNA3.1-Egr1-EGFP-Omi/HtrA2 in vitro.Method: Through liposome transfection pcDNA3.1-Egr1-EGFP was transfected into OCM-1 cells as control, pcDNA3.1-Egr1-EGFP-Omi/HtrA2 cells as the experimental group. Each group was stimulated by hydrogen peroxide. Cells in the two groups were observed by fluorescence microscopy before and after the induction. Cell proliferation was detected by MTT assay and the expression of Omi/HtrA2 and XIAP gene in the level of protein were detected by Western blotting in the control group and experimental group, apoptotic changes were observed after double-labeled with Annexin V-FITC / PI by flow cytometry.Result: Hydrogen peroxide induced expression of fluorescent cells in both groups were increased(P<0.05). Compared with the control group, cell proliferation decreased after induction, the Omi/HtrA2 gene expression at the protein level increased 53.5%, XIAP decreased 31.9%, the apoptosis of the experimental group by flow cytometry increased significantly after the induction, the difference was statistically significant (p <0.01).Conclusion: After induction the expression of plasmid transfected pcDNA3.1- Egr1-EGFP-Omi/HtrA2 can cause the apoptosis of human choroidal melanoma OCM-1 cells. It maybe due to increased expression of Omi/HtrA2, XIAP protein decrease, which advances the apoptosis.