HO-1 expression in human laryngeal squamous-cell carcinoma Hep-2 cells and its influence on Carboplatin’ s effects

LYU Xin; SONG Dong-mei; WANG Bao-shan

doi:10.3969/j.issn.1002-266X.2015.36.003

Abstract：Objective To investigate the influence of heme oxygenase-1 (HO-1) expression in human laryngeal squa-mous-cell carcinoma Hep-2 cells on the Carboplatin’ s effects and its possible mechanism.Methods Hep-2 cells in the logarithmic phase were divided into six groups.The blank control group was not treated, ZnPP group was treated with 10μmol/L HO-1 inhibitor, Hemin group was treated with 10 μmol/L HO-1 inducer, and Carboplatin group was treated with 10 μg/mL, Hemin combined with Carboplatin group was treated with 10 μmol/L Hemin alone for 1 h and then 10μg/mL Carboplatin, ZnPP combined with Carboplatin group was treated with 10 μmol/L ZnPP alone for 1 h and then 10 μg/mL Carboplatin.These six groups all had five complex holes and were cultured for 24 h after dosing.The survival rates were e-valuated by MTT assay.The expression of HO-1 mRNA and protein was detected by RT-PCR and Western blotting.Intra-cellular ROS activity was examined by DCFH-DA probe. The apoptosis rate was performed by flow cytometry. Results ①the cell survival rate:compared with the control group, the survival rates of Hep-2 cells in the ZnPP and Car-boplatin groups were significantly decreased, but the survival rate in the Hemin group was increased (all P<0.05);com-pared with the Carboplatin group, the survival rate of the Znpp combined with Carboplatin group was decreased and the He-min combined with Carboplatin group was increased (all P<0.05).②HO-1 in mRNA and protein levels:compared with the control group, the expression of HO-1 mRNA and protein was increased in the Hemin and Carboplatin groups and was decreased in the ZnPP group (all P<0.05);compared with the Carboplatin group, the Znpp combined with Carboplatin decreased, but HO-1 expression was increased in the Hemin combined with Carboplatin group (all P<0.05).③the ap-optosis rate:compared with the control group, the apoptosis rates in the Carboplatin and ZnPP groups were increased ( all P<0.05);compared with the Carboplatin group, Znpp combined with Carboplatin increased the apoptosis rate, but Hemin combined with Carboplatin decreased the apoptosis rate (all P<0.05).④ the ROS activity: compared with the control group, the the ROS activities in the Carboplatin and ZnPP groups were increased (all P<0.05);compared with the Car-boplatin group, Znpp combined with Carboplatin increased the ROS activity, but Hemin combined with Carboplatin de-creased the ROS activity (all P<0.05).Conclusions HO-1 overexpression may affect the pro-apoptotic effects of Carbo-platin on Hep-2 cells, and the application of ZnPP down-regulates the HO-1 expression and increases the apoptosis rate of Hep-2 cells with treatment of Carboplatin.In this process, the elevation of intracellular ROS activity may be involved.