Genomics & Informatics

Korea Genome Organization (한국유전체학회)
권호 표지
DOI QR코드

DOI QR Code

Characterization of H460R, a Radioresistant Human Lung Cancer Cell Line, and Involvement of Syntrophin Beta 2 (SNTB2) in Radioresistance

  • Im, Chang-Nim (Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences) ;
  • Kim, Byeong Mo (Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences) ;
  • Moon, Eun-Yi (Department of Bioscience and Biotechnology, Sejong University) ;
  • Hong, Da-Won (Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences) ;
  • Park, Joung Whan (Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences) ;
  • Hong, Sung Hee (Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences)
  • Received : 2013.09.03
  • Accepted : 2013.11.18
  • Published : 2013.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

A radioresistant cell line was established by fractionated ionizing radiation (IR) and assessed by a clonogenic assay, flow cytometry, and Western blot analysis, as well as zymography and a wound healing assay. Microarray was performed to profile global expression and to search for differentially expressed genes (DEGs) in response to IR. H460R cells demonstrated increased cell scattering and acidic vesicular organelles compared with parental cells. Concomitantly, H460R cells showed characteristics of increased migration and matrix metalloproteinase activity. In addition, H460R cells were resistant to IR, exhibiting reduced expression levels of ionizing responsive proteins (p-p53 and ${\gamma}$-H2AX); apoptosis-related molecules, such as cleaved poly(ADP ribose) polymerase; and endoplasmic reticulum stress-related molecules, such as glucose-regulated protein (GRP78) and C/EBP-homologous protein compared with parental cells, whereas the expression of anti-apoptotic X-linked inhibitor of apoptosis protein was increased. Among DEGs, syntrophin beta 2 (SNTB2) significantly increased in H460R cells in response to IR. Knockdown of SNTB2 by siRNA was more sensitive than the control after IR exposure in H460, H460R, and H1299 cells. Our study suggests that H460R cells have differential properties, including cell morphology, potential for metastasis, and resistance to IR, compared with parental cells. In addition, SNTB2 may play an important role in radioresistance. H460R cells could be helpful in in vitro systems for elucidating the molecular mechanisms of and discovering drugs to overcome radioresistance in lung cancer therapy.

Keywords

References

  1. Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010;127:2893-2917. https://doi.org/10.1002/ijc.25516
  2. Zarogoulidis K, Zarogoulidis P, Darwiche K, Boutsikou E, Machairiotis N, Tsakiridis K, et al. Treatment of non-small cell lung cancer (NSCLC). J Thorac Dis 2013;5(Suppl 4):S389- S396.
  3. Baumann M, Krause M, Hill R. Exploring the role of cancer stem cells in radioresistance. Nat Rev Cancer 2008;8:545-554. https://doi.org/10.1038/nrc2419
  4. Camphausen K, Moses MA, Beecken WD, Khan MK, Folkman J, O'Reilly MS. Radiation therapy to a primary tumor accelerates metastatic growth in mice. Cancer Res 2001;61: 2207-2211.
  5. Cheng JC, Chou CH, Kuo ML, Hsieh CY. Radiation-enhanced hepatocellular carcinoma cell invasion with MMP-9 expression through PI3K/Akt/NF-kappaB signal transduction pathway. Oncogene 2006;25:7009-7018. https://doi.org/10.1038/sj.onc.1209706
  6. Susskind H, Hymowitz MH, Lau YH, Atkins HL, Hurewitz AN, Valentine ES, et al. Increased plasma levels of matrix metalloproteinase- 9 and tissue inhibitor of metalloproteinase-1 in lung and breast cancer are altered during chest radiotherapy. Int J Radiat Oncol Biol Phys 2003;56:1161-1169. https://doi.org/10.1016/S0360-3016(03)00161-5
  7. Chou CH, Teng CM, Tzen KY, Chang YC, Chen JH, Cheng JC. MMP-9 from sublethally irradiated tumor promotes Lewis lung carcinoma cell invasiveness and pulmonary metastasis. Oncogene 2012;31:458-468. https://doi.org/10.1038/onc.2011.240
  8. Qian LW, Mizumoto K, Urashima T, Nagai E, Maehara N, Sato N, et al. Radiation-induced increase in invasive potential of human pancreatic cancer cells and its blockade by a matrix metalloproteinase inhibitor, CGS27023. Clin Cancer Res 2002; 8:1223-1227.
  9. Wong NA, Pignatelli M. Beta-catenin: a linchpin in colorectal carcinogenesis? Am J Pathol 2002;160:389-401. https://doi.org/10.1016/S0002-9440(10)64856-0
  10. Lavin MF. Ataxia-telangiectasia: from a rare disorder to a paradigm for cell signalling and cancer. Nat Rev Mol Cell Biol 2008; 9:759-769. https://doi.org/10.1038/nrm2514
  11. Satoh MS, Lindahl T. Role of poly(ADP-ribose) formation in DNA repair. Nature 1992;356:356-358. https://doi.org/10.1038/356356a0
  12. Oliver FJ, de la Rubia G, Rolli V, Ruiz-Ruiz MC, de Murcia G, Murcia JM. Importance of poly(ADP-ribose) polymerase and its cleavage in apoptosis: lesson from an uncleavable mutant. J Biol Chem 1998;273:33533-33539. https://doi.org/10.1074/jbc.273.50.33533
  13. Zhang B, Wang Y, Pang X, Su Y, Ai G, Wang T. ER stress induced by ionising radiation in IEC-6 cells. Int J Radiat Biol 2010;86:429-435. https://doi.org/10.3109/09553001003668014
  14. Zinszner H, Kuroda M, Wang X, Batchvarova N, Lightfoot RT, Remotti H, et al. CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev 1998;12:982-995. https://doi.org/10.1101/gad.12.7.982
  15. Froehner SC, Murnane AA, Tobler M, Peng HB, Sealock R. A postsynaptic Mr 58,000 (58K) protein concentrated at acetylcholine receptor-rich sites in Torpedo electroplaques and skeletal muscle. J Cell Biol 1987;104:1633-1646. https://doi.org/10.1083/jcb.104.6.1633
  16. Adams ME, Dwyer TM, Dowler LL, White RA, Froehner SC. Mouse alpha 1- and beta 2-syntrophin gene structure, chromosome localization, and homology with a discs large domain. J Biol Chem 1995;270:25859-25865. https://doi.org/10.1074/jbc.270.43.25859
  17. Bhat HF, Adams ME, Khanday FA. Syntrophin proteins as Santa Claus: role(s) in cell signal transduction. Cell Mol Life Sci 2013;70:2533-2554. https://doi.org/10.1007/s00018-012-1233-9
  18. Thames HD Jr, Withers HR, Peters LJ, Fletcher GH. Changes in early and late radiation responses with altered dose fractionation: implications for dose-survival relationships. Int J Radiat Oncol Biol Phys 1982;8:219-226. https://doi.org/10.1016/0360-3016(82)90517-X
  19. Schubert S, Knoch KP, Ouwendijk J, Mohammed S, Bodrov Y, Jager M, et al. beta2-Syntrophin is a Cdk5 substrate that restrains the motility of insulin secretory granules. PLoS One 2010;5:e12929. https://doi.org/10.1371/journal.pone.0012929
  20. Mulvey C, Harno E, Keenan A, Ohlendieck K. Expression of the skeletal muscle dystrophin-dystroglycan complex and syntrophin-nitric oxide synthase complex is severely affected in the type 2 diabetic Goto-Kakizaki rat. Eur J Cell Biol 2005;84:867-883. https://doi.org/10.1016/j.ejcb.2005.06.007
  21. Kim MJ, Froehner SC, Adams ME, Kim HS. alpha-Syntrophin is required for the hepatocyte growth factor-induced migration of cultured myoblasts. Exp Cell Res 2011;317:2914-2924. https://doi.org/10.1016/j.yexcr.2011.09.017
  22. Paglin S, Hollister T, Delohery T, Hackett N, McMahill M, Sphicas E, et al. A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res 2001;61:439-444.
  23. Lee YS, Oh JH, Yoon S, Kwon MS, Song CW, Kim KH, et al. Differential gene expression profiles of radioresistant non-small-cell lung cancer cell lines established by fractionated irradiation: tumor protein p53-inducible protein 3 confers sensitivity to ionizing radiation. Int J Radiat Oncol Biol Phys 2010;77:858-866. https://doi.org/10.1016/j.ijrobp.2009.12.076
  24. Hegedus T, Sessler T, Scott R, Thelin W, Bakos E, Varadi A, et al. C-terminal phosphorylation of MRP2 modulates its interaction with PDZ proteins. Biochem Biophys Res Commun 2003; 302:454-461. https://doi.org/10.1016/S0006-291X(03)00196-7
  25. Hoffmann U, Kroemer HK. The ABC transporters MDR1 and MRP2: multiple functions in disposition of xenobiotics and drug resistance. Drug Metab Rev 2004;36:669-701. https://doi.org/10.1081/DMR-200033473
  26. Materna V, Holm PS, Dietel M, Lage H. Kinetic characterization of ribozymes directed against the cisplatin resistance- associated ABC transporter cMOAT/MRP2/ABCC2. Cancer Gene Ther 2001;8:176-184. https://doi.org/10.1038/sj.cgt.7700293
  27. Xu QY, Gao Y, Liu Y, Yang WZ, Xu XY. Identification of differential gene expression profiles of radioresistant lung cancer cell line established by fractionated ionizing radiation in vitro. Chin Med J (Engl) 2008;121:1830-1837.
  28. Zhang X, Li X, Zhang N, Yang Q, Moran MS. Low doses ionizing radiation enhances the invasiveness of breast cancer cells by inducing epithelial-mesenchymal transition. Biochem Biophys Res Commun 2011;412:188-192. https://doi.org/10.1016/j.bbrc.2011.07.074
  29. Janikova M, Skarda J. Differentiation pathways in carcinogenesis and in chemo- and radioresistance. Neoplasma 2012; 59:6-17. https://doi.org/10.4149/neo_2012_002
  30. Woodward WA, Chen MS, Behbod F, Alfaro MP, Buchholz TA, Rosen JM. WNT/beta-catenin mediates radiation resistance of mouse mammary progenitor cells. Proc Natl Acad Sci U S A 2007;104:618-623. https://doi.org/10.1073/pnas.0606599104
  31. Yuan D, Liu L, Gu D. Transcriptional regulation of livin by beta- catenin/TCF signaling in human lung cancer cell lines. Mol Cell Biochem 2007;306:171-178. https://doi.org/10.1007/s11010-007-9567-6
  32. Artacho-Cordon F, Rios-Arrabal S, Lara PC, Artacho-Cordon A, Calvente I, Nunez MI. Matrix metalloproteinases: potential therapy to prevent the development of second malignancies after breast radiotherapy. Surg Oncol 2012;21:e143- e151. https://doi.org/10.1016/j.suronc.2012.06.001

Cited by

  1. Knockdown of PKM2 Enhances Radiosensitivity of Non-small cell Lung Cancer vol.73, pp.1, 2015, https://doi.org/10.1007/s12013-015-0567-y
  2. Evaluation of connectivity map-discovered celastrol as a radiosensitizing agent in a murine lung carcinoma model: Feasibility study of diffusion-weighted magnetic resonance imaging vol.12, pp.5, 2017, https://doi.org/10.1371/journal.pone.0178204
  3. Heat Shock Factor 1 Depletion Sensitizes A172 Glioblastoma Cells to Temozolomide via Suppression of Cancer Stem Cell-Like Properties vol.18, pp.2, 2017, https://doi.org/10.3390/ijms18020468