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Plumbagin inhibits invasion and migration of liver cancer HepG2 cells by decreasing productions of matrix metalloproteinase-2 and urokinase- plasminogen activator

Authors

  • Yuan-Wei Shih,

    1. Department of Biological Science and Technology, and Graduate Institute of Biomedical Science,
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  • Yi-Chieh Lee,

    1. Department of Biological Science and Technology, and Graduate Institute of Biomedical Science,
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  • Pei-Fen Wu,

    1. Department of Occupational Safety and Hygiene, Tajen University, Pingtung, Taiwan
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  • Yuan-Bing Lee,

    1. Department of Gynecology, Yongkang Veterans Hospital, Tainan, and
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  • Tai-An Chiang

    Corresponding author
    1. Department of Medical Technology, and Graduate Institute of Biological Science and Technology, Chung Hwa University of Medical Technology,
      Professor Tai-An Chiang, Department of Medical Technology and Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, No. 89, Wen-Hwa 1st Street, Jen-Te, Tainan 717, Taiwan. Email: giantful@mail.hwai.edu.tw
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Professor Tai-An Chiang, Department of Medical Technology and Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, No. 89, Wen-Hwa 1st Street, Jen-Te, Tainan 717, Taiwan. Email: giantful@mail.hwai.edu.tw

Abstract

Aim:  To investigate the inhibitory effects of plumbagin (5-hydroxy-2 methyl-1,4-naphthoquinone) on the invasion and migration and its correlation with matrix metalloproteinase-2 (MMP-2) and urokinase-plasminogen activator (u-PA) in liver cancer HepG2 cells under non-cytotoxic concentrations.

Methods:  The cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The adhesion, migration and invasion were measured by cell-matrix adhesion assay and Boyden chamber assay. The MMP-2 and u-PA activities were estimated by gelatin and casein-plasminogen zymography. The mRNA and protein levels of MMP-2, u-PA, urokinase-plasminogen activator receptor (u-PAR), tissue inhibitor of metalloproteinase-2 (TIMP-2), plasminogen activator inhibitor-1 (PAI-1), nuclear factor κ B (NF-κB), c-Fos and c-Jun were evaluated by semi-quantitative reverse transcription polymerase chain reaction and western blotting. Also, the binding abilities of NF-κB and activator protein-1 (AP-1) were analyzed by electrophoretic mobility shift assay (EMSA).

Results:  In this study, plumbagin had exhibited an inhibitory effect on the abilities of adhesion, migration and invasion. The results from zymography showed plumbagin treatment may decrease the activities of MMP-2 and u-PA. Further, the mRNA and protein levels of MMP-2, u-PA and u-PAR were significantly reduced, while TIMP-2 and PAI-1 were elevated by plumbagin treatment. Next, plumbagin significantly decreased the nuclear levels of NF-κB, c-Fos and c-Jun. Also, treating HepG2 cells with plumbagin leads to dose-dependent inhibition on the binding abilities of NF-κB and AP-1.

Conclusion:  We demonstrated the inhibitory effects of plumbagin on the invasion, migration and adhesion of HepG2 cells, while plumbagin treatment may decrease the expressions of MMP-2 and u-PA and enhance the expressions of TIMP-2 and PAI-1.

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