Journal of the Korean Association of Oral and Maxillofacial Surgeons

  • 제31권4호
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  • Pages.306-311
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  • 2005
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  • 2234-7550(pISSN)
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  • 2234-5930(eISSN)
대한구강악안면외과학회 (The Korean Association of Oral and Maxillofacial Surgeons)
권호 표지

재조합 HCCS-1 아데노바이러스를 이용한 구강암 세포주의 세포사멸 유발

INDUCTION OF APOPTOSIS IN ORAL CANCER CELL LINE THROUGH AN RECOMBINANT HCCS-1 ADENOVIRUS

  • 김창현 (가톨릭대학교 의과대학 치과학교실 구강악안면외과) ;
  • 이동주 (가톨릭대학교 의과대학 치과학교실 구강악안면외과) ;
  • 이일규 (가톨릭대학교 대학원 통합의학과) ;
  • 김명진 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 김진우 (가톨릭대학교 의과대학 산부인과학교실) ;
  • 표성운 (가톨릭대학교 의과대학 치과학교실 구강악안면외과)
  • Kim, Chang-Hyen (Department of Oral and Maxillofacial Surgery, College of Medicine, The Catholic University of Korea) ;
  • Lee, Dong-Ju (Department of Oral and Maxillofacial Surgery, College of Medicine, The Catholic University of Korea) ;
  • Lee, ll-Kyu (Department of Integrative Medicine, Graduate School, The Catholic University of Korea) ;
  • Kim, Myung-Jin (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Kim, Jin-Woo (Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea) ;
  • Pyo, Sung-Woon (Department of Oral and Maxillofacial Surgery, College of Medicine, The Catholic University of Korea)
  • 발행 : 2005.08.31
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초록

Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation. This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.

키워드

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