Author of

• 20039

  +

  Poster Display Session (ID 63)

  • Event: ELCC 2017
  • Type: Poster Display Session
  • Track:
  • Presentations: 1
  • Moderators:
  • Coordinates: 5/07/2017, 12:30 - 13:00, Hall 1

  • 19797

    +

    16P - Carvacrol induces growth inhibition and circumvents chemoresistance via inhibition of STAT3/Skp2/p27 pathway in non-small lung cancer cells (ID 269)

    12:30 - 13:00  |  Author(s): K. Kim
    • Abstract

    Background:
    S-phase kinase-associated protein 2 (Skp2) which constitues SCF complex and plays a role to recognize the substrates has been known to act as a proto-oncogene. In this study, we examined the effect of carvacrol, an active compound of oregano, on Skp2 inactivation and the underlying mechanisms in non-small lung cancer cells.
    
    Methods:
    Reagents and antibodies. Carvacrol was obtained from Sigma-Aldrich. A549 and H460 cells were purchased from the ATCC. For Western blot analysis, specific antibodies against Skp2, phospho-STAT3, STAT3, p21, p27, and GAPDH, as well as secondary antibodies were obtained from Santa Cruz Biotechnology.rnCell viability measurement. Cells were treated with carvacrol for 24 h and stained wtih 0.4% Trypan blue solution. Dye-excluding viable cells were counted under the microscope.rnClonogenic assay. Cells were seeded into 24 well plates and treated with carvacrol and then cultured for the next 7 to 10 days to form colonies. Colonies of > 50 cells were stained with crystal violet.rnEctopic expression of Skp2. To ectopically express Skp2, the recombinant plasmid, pcDNA3-Skp2-myc, was constructed and transfected into cells using Lipofectamine 2000. To establish stable cell lines, the transfected cells were cultured in the presence of 400 μg/ml of G418.rnsiRNA transfection. To reduce Skp2 expression, cells were transfected with siRNA targeting Skp2 or control siRNA.
    
    Results:
    Carvacrol treatment of A549 as well as H460 cells caused to reduce Skp2 protein level in dose-dependent manner. RT-PCR assay was also found that Skp2 mRNA level was reduced by carvacrol, suggesting the transcriptional down-regulation of Skp2 expression by carvacrol. We next found that the cytotoxic effect of carvacrol was attenuated in Skp2 overexpression in A459 cells and further observed its synergistic anti-proliferative effect in cells transfected with Skp2 specific siRNA. In addition, carvacrol was found to result in apoptotic cell death.
    
    Conclusions:
    Taken together, these data indicate that carvacrol targets Skp2 to inhibit cell proliferation and to cause apoptotic cell death in non-small lung cancer cells.
    
    Clinical trial identification:
    
    
    Legal entity responsible for the study:
    Yeungnam University College of Medicine, Republic of Korea
    
    Funding:
    Yeungnam University College of Medicine, Republic of Korea
    
    Disclosure:
    All authors have declared no conflicts of interest.