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

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)
Publication Information
Journal of the Korean Association of Oral and Maxillofacial Surgeons / v.31, no.4, 2005 , pp. 306-311 More about this Journal
Abstract
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.
Keywords
Oral squamous cell carcinoma; Gene therapy; Apoptosis; Human cervical cancer suppressor-1(HCCS-1); Recombinant adenovirus vector;
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1 Cusack JC, Jr, Tanabe KK: Cancer gene therapy. Surg Oncol Clin Am 1998;7:421
2 Zhang WW, Fang X, Mazur W, et al: High-efficiency gene transfer and high-level expression of wild-type p53 in human lung cancer cells mediated by recombinant adenovirus. Cancer Gene Ther 1994;1:5-13
3 김창현, 김진우, 김명진, 표성운: 구강암 유전자 치료를 위한 재조합 HCCS-1 아데노바이러스의 개발. 대악성외지 2005;27:103-109
4 Kim TE, Kim YW, Kim JW: Candidate tumor suppressor, HCCS-1, is downregulated in human cancers and induces apoptosis in cervical cancer. Int J Cancer 2002;97:780-786   DOI   ScienceOn
5 Tong-Chuan He, Shibin Zhou, Luis T, Da Costa, Jian Yu, et al: Simplified system for generating recombinant adenovirus. Proc Natl Acad Sci 1998;95:2509
6 Oltvia ZN, Milliman CL, Korsmeyer SJ: Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 1993;74:225-232
7 Jordan RC, Catzavelos GC, Barrett AW, Speight PM: Differential expression of bcl-2 and bax in squamous cell carcinoma of the oral cavity. Eur J Cancer Oral Oncol 1996;32B:394-400
8 Sherr CJ: Cancer cell cycles. Science 1996;274:1672-1677   DOI   PUBMED   ScienceOn
9 Levine AJ: the tumor suppressor gene. Annu Rev Biochem 1993;62:623-651   DOI   PUBMED   ScienceOn
10 김재우, 송재철, 이희경, 이태윤: 구강 편평세포암종 세포주 SCC-15에서 Mycolactone에 의한 apoptosis 효과. 대구외지 2001;27:511-518
11 Shillitoe EJ: Gene therapy for oral cancer; recent progress in research. Oral Oncol 1998;34:157-160   DOI   PUBMED   ScienceOn
12 Steller H: Mechanisms and genes of cellular suicide. Science 1995;267:1445-1449   DOI   PUBMED   ScienceOn
13 Schwartz GJ, Mehta RH, Wenig BL, et al: Salvage management for recurrent squamous cell carcinoma of the oral cavity. Head Neck 2000;22:34   DOI   ScienceOn
14 Chirmule N, Propert K, Magosin S, Qian Y, Qian R, Wilson J: Immune responses to adenovirus and adeno-associated virus in humans. Gene Ther 1999;6:1574-1583   DOI   ScienceOn
15 Fillis RE, Yuan J, Horvitz HR: Mechanism and function of cell death. Ann Rev Cell Biol 1991;7:663-698   DOI   ScienceOn
16 Krajewski S, Krajewska M, Shabraik A, et al: Immunohistochemical determination of in vivo distribution of Bax, a dominant inhibitors of Bbcl-2. Am J Pathol 1994;145:1323-1336
17 Wang J, Bucana CD, Roth JA et al: Apoptosis induced in human osteosarcoma cells is one of the mechanisms for the cytocidal effect of Ad5CMV-p53. Cancer Gene Ther 1995;2:9-17
18 Anderson MJ, Stanbridge EJ: Tumor suppressor genes studied by cell hybridization and chromosome transfer. FASEB J 1993;7:826-833   DOI
19 허균행, 이재훈, 김철환: 구강편평세포암종에서 미슬토 추출물의 apoptosis 유도효과에 대한 실험적 연구. 대구외지 2005;31:13-23
20 Krammer PH, Behnann I, Daniel P, et al: Regulation of apoptosis in the immune system. Curr Biol 1994;6:279-289   DOI   ScienceOn
21 Cai DW, Mukhopadhyay T, Liu Y, et al: Stable expression of the wild-type p53 gene in human lung cancer cells after retrovirus-mediated gene transfer. Hum Gene Ther 1993;4:617-624   DOI   ScienceOn
22 Kastan MB, Onyekwere O, Sidransky D, et al: Participation of p-53 protein in the cellular response to DNA damage. Cancer Res 1991;51:6304-6311