DOI QR코드

DOI QR Code

Induction of Selective Cell Death of Oral Squamous Carcinoma Cells by Integrin α2 Antibody and EGFR Antibody

인테그린 α2와 상피성장인자수용체 차단항체의 저해작용을 통한 구강편평상피암 세포의 선택적 제거

  • Choi, Yeon-Sik (Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University) ;
  • Kim, Gyoo-Cheon (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Yoon, Sik (Department of Prothodontics, School of Dentistry, Pusan National University) ;
  • Hwang, Dae-Seok (Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University) ;
  • Kim, Cheol-Hun (Department of Oral and Maxillofacial Surgery, College of Medicine, Dong-A University) ;
  • Jeon, Young-Chan (Department of Prothodontics, School of Dentistry, Pusan National University) ;
  • Byun, June-Ho (Department of Oral and Maxillofacial Surgery, School of Medicine, Gyeongsang National University) ;
  • Shin, Sang-Hun (Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University) ;
  • Kim, Uk-Kyu (Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University)
  • 최연식 (부산대학교 치의학전문대학원 구강악안면외과학교실) ;
  • 김규천 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 윤식 (부산대학교 치의학전문대학원 보철학교실) ;
  • 황대석 (부산대학교 치의학전문대학원 구강악안면외과학교실) ;
  • 김철훈 (동아대학교 의과대학 구강악안면외과학교실) ;
  • 전영찬 (부산대학교 치의학전문대학원 보철학교실) ;
  • 변준호 (경상대학교 의학전문대학원 구강악안면외과학교실) ;
  • 신상훈 (부산대학교 치의학전문대학원 구강악안면외과학교실) ;
  • 김욱규 (부산대학교 치의학전문대학원 구강악안면외과학교실)
  • Received : 2013.02.20
  • Accepted : 2013.05.14
  • Published : 2013.05.31

Abstract

Purpose: This study was to find efficacy of integrin alpha2 (${\alpha}_2$) and epidermal growth factor receptor (EGFR) as tumor marker of oral squamous cell carcinoma (SCC) and clarify the selective cell death effect of anti-integrin ${\alpha}_2$ and anti-EGFR on SCC cells, additionally testify conjugated gold nanoparticles (GNP) with air plasma for selective cell death of oral SCC. Methods: Expression of integrin ${\alpha}_2$, EGFR on human SCC cells (SCC25) were examined by western blot. SCC25 cells were treated with anti-integrin ${\alpha}_2$, anti-EGFR and analysed by Hemacolor staining, immunoflorescence staining, FACS flow cytometry. Conjugated GNP with integrin ${\alpha}_2$, EGFR antibody were treated by air plasma on SCC cells. Results: Integrin ${\alpha}_2$ and EGFR were over-expressed on SCC25 cells than normal lung WI-38 cells. The cell viability rate of SCC25 cells treated with anti-integrin ${\alpha}_2$, anti-EGFR was lower than WI-38 cells. The concentration changes of nucleus, releasing cytochrome c and apoptosis inducing factor (AIF) from mitochondria to cytosol were observed. The changes of proteins related with apoptosis were observed. Increase of bax, bcl-xL, activation of caspase-3, -7, -9, and fragmentation of PARP, DFF45 and decrease of lamin A/C in SCC25 cells were observed. In FACS, increase of sub-$G_1$ and S phase was observed. Cell cycle related proteins, Such as cyclin D1, cyclin dependent kinase (CDK) 4, cyclin A, cyclin E, CDK 2, p27 were decreased. After SCC25 cells treated with conjugatged GNP-Integrin ${\alpha}_2$, GNP-EGFR, additionally air plasma, the cell death rate was significantly increased. Conclusion: Integrin ${\alpha}_2$, EGFR were over-expressed in oral SCC cells. Anti-integrin ${\alpha}_2$, anti-EGFR in SCC25 cells induced apoptosis selectively. When GNP-anti integrin ${\alpha}_2$, GNP-anti EGFR were treated with air plasma on SCC25 cells, cancer cells were died more selectively. GNP-anti integrin ${\alpha}_2$, GNP-anti EGFR with air plasma could be treatment choice of oral SCC.

Keywords

References

  1. Hughes AL. Evolution of the integrin alpha and beta protein families. J Mol Evol 2001;52:63-72. https://doi.org/10.1007/s002390010134
  2. Guo W, Giancotti FG. Integrin signalling during tumour progression. Nat Rev Mol Cell Biol 2004;5:816-26. https://doi.org/10.1038/nrm1490
  3. Guo L, Zhang F, Cai Y, Liu T. Expression profiling of integrins in lung cancer cells. Pathol Res Pract 2009;205:847-53. https://doi.org/10.1016/j.prp.2009.07.005
  4. Bargmann CI, Hung MC, Weinberg RA. The neu oncogene encodes an epidermal growth factor receptor-related protein. Nature 1986;319:226-30. https://doi.org/10.1038/319226a0
  5. Herbst RS. Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys 2004;59(2 Suppl):21-6. https://doi.org/10.1016/j.ijrobp.2003.10.027
  6. Ishitoya J, Toriyama M, Oguchi N, et al. Gene amplification and overexpression of EGF receptor in squamous cell carcinomas of the head and neck. Br J Cancer 1989;59:559-62. https://doi.org/10.1038/bjc.1989.113
  7. Neve RM, Lane HA, Hynes NE. The role of overexpressed HER2 in transformation. Ann Oncol 2001;12 Suppl 1:S9-13. https://doi.org/10.1093/annonc/12.suppl_1.S9
  8. Ullrich A, Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell 1990;61:203-12. https://doi.org/10.1016/0092-8674(90)90801-K
  9. Young NR, Liu J, Pierce C, et al. Molecular phenotype predicts sensitivity of squamous cell carcinoma of the head and neck to epidermal growth factor receptor inhibition. Mol Oncol 2012. pii: S1574-7891(12)00121-4.
  10. Markovic A, Chung CH. Current role of EGF receptor monoclonal antibodies and tyrosine kinase inhibitors in the management of head and neck squamous cell carcinoma. Expert Rev Anticancer Ther 2012;12:1149-59. https://doi.org/10.1586/era.12.91
  11. Kim KW, Kim MJ. Expression of the epidermal growth factor receptor and cell cycle analysis in the head and neck squamous cell carcinomas. J Korean Assoc Oral Maxillofac Surg 2000;26:154-63.
  12. Moon BC, Han SJ, Jeong D, Kim KW. Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011;37:396-402. https://doi.org/10.5125/jkaoms.2011.37.5.396
  13. Fridman G, Fridman G, Gutsol A, Shekhter AB, Vasilets VN, Fridman A. Applied plasma medicine. Plasma Process Polym 2008;5:503-33. https://doi.org/10.1002/ppap.200700154
  14. Fridman G, Shereshevsky A, Jost MM, et al. Floating elctrode dielectric barrier discharge plasma in air promoting apoptotic behavior in melanoma skin cancer cell lines. Plasma Chem Plasma Process 2007;27:163-76. https://doi.org/10.1007/s11090-007-9048-4
  15. Kim CH, Kwon S, Bahn JH, et al. Effects of atmospheric nonthermal plasma on invasion of colorectal cancer cells. Appl Phys Lett 2010;96:243701. https://doi.org/10.1063/1.3449575
  16. Kim D, Gweon B, Kim DB, Choe W, Shin JH. A feasibility study for the cancer therapy using cold plasma. ICBME Proc 2008;23:355-7.
  17. Kim GC, Lee HJ, Shon H. The effects of a micro plasma on melanoma (G361) cancer cells. J Korean Phys Soc 2009;54:628-32. https://doi.org/10.3938/jkps.54.628
  18. Connor EE, Mwamuka J, Gole A, Murphy CJ, Wyatt MD. Gold nanoparticles are taken up by human cells but do not cause acute cytotoxicity. Small 2005;1:325-7. https://doi.org/10.1002/smll.200400093
  19. Thomas M, Klibanov AM. Conjugation to gold nanoparticles enhances polyethylenimine's transfer of plasmid DNA into mammalian cells. Proc Natl Acad Sci U S A 2003;100:9138-43. https://doi.org/10.1073/pnas.1233634100
  20. Levy JM, Thorburn A. Targeting autophagy during cancer therapy to improve clinical outcomes. Pharmacol Ther 2011;131:130-41. https://doi.org/10.1016/j.pharmthera.2011.03.009
  21. Chen CS, Alonso JL, Ostuni E, Whitesides GM, Ingber DE. Cell shape provides global control of focal adhesion assembly. Biochem Biophys Res Commun 2003;307:355-61. https://doi.org/10.1016/S0006-291X(03)01165-3
  22. Riveline D, Zamir E, Balaban NQ, et al. Focal contacts as mechanosensors: externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism. J Cell Biol 2001;153:1175-86. https://doi.org/10.1083/jcb.153.6.1175
  23. Zamir E, Geiger B. Molecular complexity and dynamics of cell-matrix adhesions. J Cell Sci 2001;114:3583-90.
  24. Ennis BW, Lippman ME, Dickson RB. The EGF receptor system as a target for antitumor therapy. Cancer Invest 1991;9:553-62. https://doi.org/10.3109/07357909109018953
  25. Mendelsohn J, Baselga J. Epidermal growth factor receptor targeting in cancer. Semin Oncol 2006;33:369-85. https://doi.org/10.1053/j.seminoncol.2006.04.003
  26. Ghavami S, Hashemi M, Ande SR, et al. Apoptosis and cancer: mutations within caspase genes. J Med Genet 2009;46:497-510. https://doi.org/10.1136/jmg.2009.066944
  27. Dejean LM, Martinez-Caballero S, Manon S, Kinnally KW. Regulation of the mitochondrial apoptosis-induced channel, MAC, by BCL-2 family proteins. Biochim Biophys Acta 2006;1762:191-201. https://doi.org/10.1016/j.bbadis.2005.07.002
  28. Susin SA, Lorenzo HK, Zamzami N, et al. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 1999;397:441-6. https://doi.org/10.1038/17135
  29. Dejean LM, Martinez-Caballero S, Kinnally KW. Is MAC the knife that cuts cytochrome c from mitochondria during apoptosis? Cell Death Differ 2006;13:1387-95. https://doi.org/10.1038/sj.cdd.4401949
  30. Alnemri ES, Livingston DJ, Nicholson DW, et al. Human ICE/CED-3 protease nomenclature. Cell 1996;87:171. https://doi.org/10.1016/S0092-8674(00)81334-3
  31. Murphy KM, Ranganathan V, Farnsworth ML, Kavallaris M, Lock RB. Bcl-2 inhibits Bax translocation from cytosol to mitochondria during drug-induced apoptosis of human tumor cells. Cell Death Differ 2000;7:102-11. https://doi.org/10.1038/sj.cdd.4400597
  32. Yu SW, Andrabi SA, Wang H, et al. Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. Proc Natl Acad Sci U S A 2006;103:18314-9. https://doi.org/10.1073/pnas.0606528103
  33. Widłak P. The DFF40/CAD endonuclease and its role in apoptosis. Acta Biochim Pol 2000;47:1037-44.
  34. Karam JA. Apoptosis in carcinogenesis and chemotherapy. Netherlands: Springer; 2009.
  35. Morgan DO. Principles of CDK regulation. Nature 1995;374:131-4. https://doi.org/10.1038/374131a0
  36. Sherr CJ. G1 phase progression: cycling on cue. Cell 1994;79:551-5. https://doi.org/10.1016/0092-8674(94)90540-1
  37. Pines J. Cyclins: wheels within wheels. Cell Growth Differ 1991;2:305-10.
  38. Si X, Liu Z. Expression and significance of cell cycle-related proteins Cyclin DI, CDK4, p27, E2F-I and Ets-1 in chondrosarcoma of the jaws. Oral Oncol 2001;37:431-6. https://doi.org/10.1016/S1368-8375(00)00091-9
  39. Kim GC, Kim GJ, Park SR, et al. Air plasma coupled with antibody-conjugated nanoparticles: a new weapon against cancer. J Phys D Appl Phys 2009;42:032005. https://doi.org/10.1088/0022-3727/42/3/032005
  40. Choi BB, Choi YS, Lee HJ, Lee JK, Kim UK, Kim GC. Nonthermal plasma-mediated cacer cell death: targeted cancer treatment. J Therm Sci Technol 2012;7:399-404. https://doi.org/10.1299/jtst.7.399