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Suppressive effects of genomic imprinted gene PEG10 on hydrogen peroxide-induced apoptosis in L02 cells

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Summary

The effects of PEG10 on hydrogen peroxide (H2O2)-induced apoptosis in human normal liver cell line L02 were investigated. The PEG10 gene was transfected into L02 cells by lipofectamine, the positive clone was screened by G418 and defined as L02/PEG10, while the cell transfected with empty expression vector (pEGFP-N1) was defined as L02/vector. L02/vector and parental L02 cells served as control. RT-PCR and Western blotting were employed to detect the expression of target genes. H2O2 (50–400 mmol/L) was administered to induce the apoptosis of L02 cells. Cells viability was measured by MTT and the morphological changes of apoptotic cells were determined by fluorescence microscopy using hoechst33342 nuclei staining. DNA fragmentation was observed by agarose gel electrophoresis. PEG10 mRNA and protein levels in L02/PEG10 cells were significantly increased as compared with those in the control cells. After treatment with 400 mmol/L H2O2 for 24 h, the cellular growth inhibition rate of L02/PEG10 cells was significantly lower (58.2%) than that of L02 (92.5%) and L02/vector (88%). Distinct morphological changes characteristic of cell apoptosis such as karyopyknosis and conglomeration were not observed in L02/PEG10. Ladder-like DNA fragmentation in a dose-dependent manner was observed in both L02 and L02/vector cell lines, but not in L02/PEG10. PEG10 over-expression significantly inhibited cytotoxicity induced by H2O2 on human normal liver cell line L02 by antagonizing H2O2-induced apoptosis.

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Authors and Affiliations

  1. Department of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Yao Liu  (刘 瑶), Jusheng Lin  (林菊生) & Qiang Zhang  (张 强)

  2. Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Huanjun Huang  (黄焕军)

  3. Department of Immunology, School of Medicine, Wuhan University, Wuhan, 430071, China

    Jinquan Tan  (谭锦泉)

  4. Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430023, China

    Jinghua Ren  (任精华)

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  1. Yao Liu  (刘 瑶)
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  2. Huanjun Huang  (黄焕军)
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  3. Jusheng Lin  (林菊生)
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  4. Qiang Zhang  (张 强)
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  5. Jinquan Tan  (谭锦泉)
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  6. Jinghua Ren  (任精华)
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Additional information

This project was supported by grants from National Natural Sciences Foundation of China (No. 30471983, 30872237) and Major State Basic Research Development Program of China (973 Program) (No. 2007CB512900).

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Liu, Y., Huang, H., Lin, J. et al. Suppressive effects of genomic imprinted gene PEG10 on hydrogen peroxide-induced apoptosis in L02 cells. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 29, 705–709 (2009). https://doi.org/10.1007/s11596-009-0606-2

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  • DOI: https://doi.org/10.1007/s11596-009-0606-2

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