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C. Park



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    P1.04 - Poster Session/ Biology, Pathology, and Molecular Testing (ID 233)

    • Event: WCLC 2015
    • Type: Poster
    • Track: Biology, Pathology, and Molecular Testing
    • Presentations: 1
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      P1.04-048 - DNA Demethylation Related Hypoxia-Induced Stem-Like Properties in Lung Cancer (ID 614)

      09:30 - 09:30  |  Author(s): C. Park

      • Abstract
      • Slides

      Background:
      Tumors exhibiting extensive hypoxia have been shown to be more aggressive than corresponding tumors that are better oxygenized, which suggests that hypoxic condition induces stem-like properties. The purpose of the present study was to investigate whether hypoxic stress induces acquisition of stem-like properties, and the mechanism is involved with DNA demethylation in lung cancer.

      Methods:
      Normal epithelial cell line (BEAS-2B) and human lung cancer cell lines (A549, H292, H226 and H460) were incubated either in normoxic or in hypoxic (below 1% O~2~) condition. The cell lines were treated with a DNA methyltransferase inhibitor (5-azacytidine, AZA) to determine whether the expression of stem cell markers (CD44, CD133, CXCR4, ABCG2, CD117, ALDH1A1, EpCAM, CD90, Oct4, Nanog, SOX2, SSEA4 and CD166) was reactivated. Methylation-specific PCR and bisulfite sequencing were used to analyze the methylation status, and real-time RT-PCR and western blotting were performed to analyze the expression of the stem cell markers. Cell migration and Matrigel invasion assay were performed for functional analysis

      Results:
      Among the 13 stem cell markers, CXCR4, Oct4 and Nanog were increased at least one lung cancer cell line in hypoxic condition compared with in normoxic condition. These three stem cell markers were reactivated by treatment with AZA. Methylation-specific PCR showed decreased promoter methylation of these three stem cell markers in hypoxic condition compared with in normoxic condition, which was further validated by bisulfite sequencing. Migration and invasion were increase in hypoxic condition compared with in normoxic condition

      Conclusion:
      These results suggest that under the hypoxic condition, reactivation of stem-like properties was related with promoter demethylation of stem cell markers. Further studies are needed to assess its value as a prognostic factor and potential therapeutic applications.

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