Earticle

초록

영어

Background: Tumor response to anticancer therapies can much be influenced by microenvironmental factors. In this study, we determined the effect of these microenvironmental factors on DNA methylation using A549 human lung adenocarcinoma cell line. Materials and Methods: We subjected A549 cells to various conditions mimicking tumor microenvironment including hypoxia, acidosis (sodium lactate), oxidative stress (H2O2), bystander effect (supernatant from doxorubicin (Dox)-treated or irradiated cells), and immune cell infiltration (supernatant from THP-1 or Jurkat T cells). Genomic DNA was isolated from these cells and analyzed for DNA methylation. Clonogenic cell survival, gene expression, and metabolism were analyzed in cells treated with some of these conditions. Results and Discussion: We found that DNA methylation level was significantly decreased in A549 cells treated with conditioned media from Dox-treated cells or Jurkat T cells, or sodium lactate, indicating an active transcription. To determine whether the decreased DNA methylation affects radiation sensitivity, we exposed cells to these conditions followed by 6 Gy irradiation and found that cell survival was significantly increased by sodium lactate while it was decreased by conditioned media from Dox-treated cells. We further observed that cells treated with conditioned media from Dox-treated cells exhibited significant changes in expression of genes including BAX and FAS (involved in apoptosis), NADPH dehydrogenase (mitochondria), EGFR (cellular survival) and RAD51 (DNA damage repair) while sodium lactate increased cellular metabolism rather than changing the gene expression. Conclusion: Our results suggest that various tumor microenvironmental factors can differentially influence DNA methylation and hence radiosensitivity and gene expression in A549 cancer cells.

목차

ABSTRACT
 Introduction
 Materials and Methods
  1. Cell culture and treatments
  2. Cell viability
  3. Methylated DNA Quantification
  4. Ionizing radiation and cell surviving fraction
  5. RNA extraction and cDNA synthesis
  6. Quantitative real time-polymerase chain reaction(qRT-PCR)
  7. Seahorse cellular metabolism measurements
  8. Statistics
 Results
  1. Effects of tumor microenvironmental factors on DNA methylation in A549 cells
  2. Effects of DNA methylation on radiosensitivity of A549 cells
  3. Dox-treated conditioned media increase gene expression pathways involving apoptosis and DNA repair while lactate modulates cellular metabolism to affect radiosensitivity
 Discussion
 Acknowledgements
 References

키워드

저자

• Jung-Min Oh [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Young-Eun Kim [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Beom-Ju Hong [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Seoyeon Bok [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Seong-Uk Jeon [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Chan-Ju Lee [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Dong-Young Park [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea ]
• Il Han Kim [ Department of Radiation Oncology, College of Medicine, Seoul National University, Seoul, Korea, Cancer Research Institute, College of Medicine, Seoul National University, Seoul, Korea; ,Institute of Radiation Medicine, Medical Research Center, Seoul National University, Seoul, Korea ]
• Hak Jae Kim [ Department of Radiation Oncology, College of Medicine, Seoul National University, Seoul, Korea, Cancer Research Institute, College of Medicine, Seoul National University, Seoul, Korea; ,Institute of Radiation Medicine, Medical Research Center, Seoul National University, Seoul, Korea ] Co-corresponding author
• G-One Ahn [ Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, Korea, Department of Life Science, Pohang University of Science and Technology Pohang, Korea ] Corresponding autho

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  방사선방어학회지 [Journal of Radiation Protection and Research]

• 간기

  계간

• pISSN

  2508-1888

• 수록기간

  1976~2026

• 등재여부

  KCI 등재,SCOPUS

• 십진분류

  KDC 559 DDC 629