Transfection of Ad-HO-1 on renal tubular epithelial cell HK-2 and its effect on PBMC proliferation / 第三军医大学学报

ABSTRACT

Objective To observe the cytotoxicity of Ad-HO-1 and its capacity of mediating the expression of HO-1 in cultured HK-2 cells. Methods The HK-2 cells were infected by Ad-HO-1 with 100, 200 and 400 MOI for 3 h, followed by the green fluorescence observation 2 days later. The live cells ratio was detected 2 and 4 days later. The expression of HO-1 and GFP in HK-2 were detected under laser scan confocal microscope. The expression of HO-1 was detectee by Western blot analysis. 3H-TdR was used to assay the ability of HO-2 cells infected with Ad-HO-1 to inhibit the PBMC proliferation. Results Over 90% HK-2 cells expressed green fluorescence after infected by Ad-HO-1 (MOI 200) for 2 days. The live cell ratio at 2 and 4 days had not any difference from those of the control group. The expressions of HO-1 and GFP in the Ad-HO-1 transfected HK-2 cells were determined under laser scan confocal microscope. The locations of these two proteins were the same. HK-2 cells infected by Ad-HO-1 had protein immune blot strap combined with HO-1 antibody. When the MOI of Adv-HO-1 was 0.01, 0.1, 1 and 10, the proliferative capacity of PBMC was inhibited significantly. Conclusion The stable expression of HO-1 mediated by Ad-HO-1 in cultured tubular epithelial cells can inhibit the cell proliferation of PBMC.