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Increased Expression of ILP-2 Alleviate Apoptosis Induced by H2O2 in Human MCF-7 Cells
Zhu Lin1, Wang Siyuan1, Xiang Siqi2, Zhou Peiyun1, Wang Yani1, Zhu Liu1, Huang Huiqun1, Xiang Mingjun1*
1Department of Biochemistry and Immunology, Medical Research Center, Institute of Medicine, Jishou University, Jishou 416000, China; 2Department of Bioengineering, Biological Science and Engineering School, North University of Nationalities, Yinchuan 750021, China
Abstract: The aim of this paper was to investigate the effect of ILP-2 (inhibitor of apoptosis protein-like protein-2) on reactive oxygen species (ROS) oxidative stress in breast cancer cells. Oxidative stress model on MCF- 7 cells was established by different concentrations of H2O2. DCFH-DA probe was applied to detect ROS in breast cancer line. When small interfering RNA (siRNA) of ILP-2 was designed to transfect into the MCF-7 cells， the expression of ILP-2 was measured by Western blot. Additionally， the breast cancer line treated by the siRNA and H2O2， the proliferation activity of MCF-7 cell was analyzed by MTT method. Cell scratch test was performed to examine the migration effects of siRNA-transfected and hydrogen peroxide treatment MCF-7 cell. The apoptosis effects of siRNA-transfected and H2O2 treated on MCF-7 cell was detected by AO-EB double staining method. The results demonstrated that 200 μmol/L H2O2 significantly instigated the production of ROS in the breast cancer cell line MCF-7. Western blot results showed that siRNA-5 had the interference higher efficiency， what’s more， the expression of ILP-2 was higher in H2O2+siRNA-5 group than H2O2+siRNA-NC group. After treatment with H2O2 for 24， 48 and 72 hours， MTT assay results showed that cell survival rate was significantly lower than those in the control group， the H2O2+siRNA-5 group’s was higher than those in the H2O2+siRNA-NC group’s. The migration rate decreased compared with the control group， and the migration rate of the H2O2+siRNA-5 group increased than the H2O2+siRNA-NC group’s. AO-EB results showed that compared with the control group， the cell apoptosis rate of the H2O2 group was obviously increased， and the H2O2+siRNA-5 group’s was decreased than that of the H2O2+siRNA-NC. The apoptosis inhibitory protein ILP-2 can resist the cytotoxicity of ROS in breast cancer cells MCF-7 and promoted cell proliferation.
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