Anti-metastatic mechanism of mountain cultivated wild ginseng in human cancer cell line

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  • ABSTRACT

    Objective

    Ginseng is one of most widely used herbal medicine. Ginseng showed anti-metastasis activities. However, its molecular mechanisms of action are unknown. So we want to report the wild ginseng repress which plays key roles in neoplastic epithelial-mesenchymal transition process.

    Methods

    Treatment of the human colorectal carcinoma LOVO cells and human gastric carcinoma SNU601 cells with the increased concentrations of cultivated wild ginseng extracts resulted in a gradual decrease in the AXIN2 gene expression.

    Results

    Metastasis-suppressor genes, maspin and nm23 was not affected by the treatment of ginseng extracts in LOVO cells. Moreover, the mountain cultivated wild ginseng or mountain wild ginseng are similar in their inhibitory effects on the expression of AXIN2 gene, but are substantially stronger than cultivated ginseng.

    Conclusion

    We described the novel mechanism of wild ginseng-induced anti-metastasis activity by repressing the expression of AXIN2 gene that plays key roles in epithelial-mesenchymal transition process.


  • KEYWORD

    Ginseng , cultivated wild ginseng , antimetastasis activities , AXIN2 gene

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  • [Fig. 1] Cultivated ginseng(CG-a), mountain cultivated wild ginseng(MCWG-b) and mountainwild ginseng(WG-c) wereused for ginseng extracts.
    Cultivated ginseng(CG-a), mountain cultivated wild ginseng(MCWG-b) and mountainwild ginseng(WG-c) wereused for ginseng extracts.
  • [Fig. 2] Schematic diagram showing that β?-catenin,stabilized either by canonical Wnt signalling,triggers TCF-dependent signaling that inducesAxin2 expression. In turn, Snail1 protein half-lifeis increased, thereby engaging a Snail1-dependentEMT process.
    Schematic diagram showing that β?-catenin,stabilized either by canonical Wnt signalling,triggers TCF-dependent signaling that inducesAxin2 expression. In turn, Snail1 protein half-lifeis increased, thereby engaging a Snail1-dependentEMT process.
  • [Fig. 3] Repression of AXIN2 mRNA after treating LOVO andSNU601 cells with MCWG. AXIN2 mRNA levels wereanalyzed by semi-quantitative RT-PCR. Human colorectalcarcinoma LOVO cells and human gastric carcinomaSNU601 cells were cultured in media containingmountain cultivated wild ginseng extracts (0 -100μg/ml) for 3 days, and the AXIN2 mRNA level wasevaluated by semi-quantitative RT-PCR using the AXIN2-specific primers. RT-PCR analysis revealedthe expected 325 bp AXIN2 band and β-actin being aninternal control.
    Repression of AXIN2 mRNA after treating LOVO andSNU601 cells with MCWG. AXIN2 mRNA levels wereanalyzed by semi-quantitative RT-PCR. Human colorectalcarcinoma LOVO cells and human gastric carcinomaSNU601 cells were cultured in media containingmountain cultivated wild ginseng extracts (0 -100μg/ml) for 3 days, and the AXIN2 mRNA level wasevaluated by semi-quantitative RT-PCR using the AXIN2-specific primers. RT-PCR analysis revealedthe expected 325 bp AXIN2 band and β-actin being aninternal control.
  • [Fig. 4] Quantitative real-time RT-PCR analysis of AXIN2 transcripts.Total RNA extracted from the cells (2μg) wasreverse-transcribed to cDNA (40μl), and aliquots (1μl)were applied to real-time PCR(20μl) with eachprimer(0.4 mM). Values represented relative expression of AXIN2 gene (calculated with threshold cyclenumber, CT) of MCWG treated cells compared withthat of non-treated control cells. Each value wasadjusted with CT of internal control(β-actin).
    Quantitative real-time RT-PCR analysis of AXIN2 transcripts.Total RNA extracted from the cells (2μg) wasreverse-transcribed to cDNA (40μl), and aliquots (1μl)were applied to real-time PCR(20μl) with eachprimer(0.4 mM). Values represented relative expression of AXIN2 gene (calculated with threshold cyclenumber, CT) of MCWG treated cells compared withthat of non-treated control cells. Each value wasadjusted with CT of internal control(β-actin).
  • [Fig. 5] RT-PCR analysis of AXIN2 mRNA expression inhuman colorectal carcinoma LOVO cell linestreated with the CG extracts, MCWG extracts, or WG extracts(100㎍/㎖) for 3 days. Lane (-), negativecontrol.
    RT-PCR analysis of AXIN2 mRNA expression inhuman colorectal carcinoma LOVO cell linestreated with the CG extracts, MCWG extracts, or WG extracts(100㎍/㎖) for 3 days. Lane (-), negativecontrol.
  • [Fig. 6] Expression of mRNA for maspin and nm23 on LOVO cells. Total RNA(2μg) from LOVO cells treated with the CG extracts, MCWG extracts, or WG extracts(100㎍/㎖) for 3 days was examined by RTPCR analysis. Lane (-), negative control.
    Expression of mRNA for maspin and nm23 on LOVO cells. Total RNA(2μg) from LOVO cells treated with the CG extracts, MCWG extracts, or WG extracts(100㎍/㎖) for 3 days was examined by RTPCR analysis. Lane (-), negative control.
  • [Table 1] Primer for RT-PCR
    Primer for RT-PCR