Journal of Oral Medicine and Pain

Korean Academy of Orofacial Pain and Oral Medicine (대한안면통증구강내과학회)
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Identification of Genes Connected with the Sensitivity to 5-FU and Cisplatin in Squamous Cell Carcinoma Cell Lines

편평세포암 세포주에서 5-FU와 Cisplatin에의 감수성과 관련된 유전자의 동정

  • Choi, Na-Young (Dept. of Oral Medicine, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Kim, Ok-Joon (Dept. of Oral Pathology, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Lee, Geum-Sug (Dept. of Oral Medicine, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Kim, Byung-Gook (Dept. of Oral Medicine, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Kim, Jae-Hyeong (Dept. of Oral Medicine, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Jang, Youn-Young (Dept. of Oral Pathology, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Lim, Won-Bong (Dept. of Oral Pathology, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Chong, Min-A (Dept. of Oral Pathology, School of Dentistry, Dental Science Research Institute, Chonnam National University) ;
  • Choi, Hong-Ran (Dept. of Oral Pathology, School of Dentistry, Dental Science Research Institute, Chonnam National University)
  • 최나영 (전남대학교 치의학전문대학원 구강내과학교실 전남대학교 치의학연구소) ;
  • 김옥준 (전남대학교 치의학전문대학원 구강병리학교실 전남대학교 치의학연구소) ;
  • 이금숙 (전남대학교 치의학전문대학원 구강내과학교실 전남대학교 치의학연구소) ;
  • 김병국 (전남대학교 치의학전문대학원 구강내과학교실 전남대학교 치의학연구소) ;
  • 김재형 (전남대학교 치의학전문대학원 구강내과학교실 전남대학교 치의학연구소) ;
  • 장윤영 (전남대학교 치의학전문대학원 구강병리학교실 전남대학교 치의학연구소) ;
  • 임원봉 (전남대학교 치의학전문대학원 구강병리학교실 전남대학교 치의학연구소) ;
  • 정민아 (전남대학교 치의학전문대학원 구강병리학교실 전남대학교 치의학연구소) ;
  • 최홍란 (전남대학교 치의학전문대학원 구강병리학교실 전남대학교 치의학연구소)
  • Published : 2005.09.30
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Abstract

Squamous cell carcinoma (SCC) in head and neck show a variability in the response to chemotherapy, even when it present with similar histological tumor type, grade, and clinical stage. The purpose of present study it to identify predictive bio-marker for the sensitivity or resistance to conventional chemotherapeutic agents, 5-fluorouracil (5-FU) and Cisplatin Oral cancer cell lines were used in present study. MTT assay was performed to evaluate the sensitivity and/or resistance to 5-FU and Cisplatin. And RT-PCR was carried out for evaluation of the mRNA expressions of various genes associated with mutation, inflammation (COX pathway), cell cycle, senescence and extracellular matrix (ECM). The molecules which are correlated with the sensitivity to 5-FU are XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 and HSP47. And the molecules are correlated with the sensitivity to Cisplatin are COX-1, iNOS, eNOS, PCNA, collagen 1 and MMP-9. Taken together, when choosing the appropriate chemotherpeutic agents for patients, considering the molecules which are correlated or reversely correlated is helpful to choose the resonable agents for cancer patients.

두경부에서 발생하는 편평세포암종은 같은 조직학적 분류, 조직학적 단계 및 임상 단계에 있다하더라도 항암제 투여에 따른 감수성은 환자마다 다양하게 나타난다. 따라서, 본 연구의 목적은 기존에 가장 많이 사용하는 항암제인 5-FU 와 Cisplatin의 감수성을 예측할 수 있는 생물학적 표지자를 찾아 환자에게 맞는 항암제를 선택하기 위해서이다. 5종의 구강 편평암세포암주를 이용하였으며, 5-FU 와 Cisplatin의 항암제 감수성을 측정하기 위해 MTT assay를 시행하였다. 각 세포 주의 전 RNA를 추출하였으며 이어서 cDNA를 합성하였다. 다양한 유전자를 1) 돌연변이 2) 염증(COX pathway) 3) 세포주기 4) 노화 5) 세포외기질 과 관련되게 분류하였고 RT-PCR을 시행하여 유전자의 발현량을 살펴보았다. 결과물은 Scion image 프로그램을 통해 분석하였고, Sigma plot을 이용해 표시하였다. 5-FU의 감수성과 비례하는 유전자들은 XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 및 HSP47이었으며, Cisplatin의 감수성과 비례하는 유전자들은 COX-1, iNOS, eNOS, PCNA, Col-1 및 MMP-9 으로 알 수 있었다. 위 결과는 암환자에게 알맞은 항암제를 선택 시 항암제의 감수성과 관련한 유전자들의 발현 정도를 파악한다면 올바른 항암제를 선택하는데 도움이 되리라 사료된다.

Keywords

References

  1. Franceschi S, Barra S, Vecchia C, Bidoli E, Negri E, and Talamini R. Risk factors for cancer of the tongue and the mouth. A case control study from northern Italy. Cancer 1992;70:2227-2233 https://doi.org/10.1002/1097-0142(19921101)70:9<2227::AID-CNCR2820700902>3.0.CO;2-Z
  2. Field JK, Kiaris H, Risk JM, et al. Allelotype of squamous cell carcinoma of the head and neck fractional allele loss correlates with survival. Br J Cancer 1995;72:1180-1188 https://doi.org/10.1038/bjc.1995.483
  3. Paccagnella A, Orlando A, Marchiori C, et al. Phase III trial of initial chemotherapy in stage III or IV head and neck cancers: a study by the Gruppo di Studio sui Tumori della Testa e del Collo. J Natl Cancer Inst. 1994;16;86:265-272
  4. Pignon JP, Bourhis J, Domenge C, and Designe L. Chemotherapy added to locoregional treatment for head and neck squamous-cell carcinoma: three meta-analyses of updated individual data. MACH-NC Collaborative Group. Meta-Analysis of Chemotherapy on Head and Neck Cancer. Lancet. 2000;18;355:949-955
  5. Lefebvre JL, Chevalier D, Luboinski B, Kirkpatrick A, Collette L, Sahmoud T. Larynx preservation in pyriform sinus cancer: preliminary results of a European Organization for Research and Treatment of Cancer phase III trial. EORTC Head and Neck Cancer Cooperative Group. J Natl Cancer Inst. 1996;3;88:890-899
  6. Veterans Affairs Laryngeal Cancer Study Group. Induction chemotherapy plus radiation compared with surgery plus radiation in patients with advanced laryngeal cancer. N Engl J Med. 1991; 13;324(24): 1685-1690
  7. Zaffaroni N, Silvestrini R, Orlandi L, Bearzatto A, Gornati D, and Villa R. Induction of apoptosis by taxol and cisplatin and effect on cell cycle-related proteins in cisplatin-sensitive and -resistant human ovarian cells. Br J Cancer. 1998;77:1378-1385 https://doi.org/10.1038/bjc.1998.230
  8. Park JS, Yoon SY, Kim JM, Yeom YI, Kim YS, and Kim NS. Identificatin of novel genes associated with the response to 5-FU treatment in gastric cancer cell lines using a cDNA microarray. Cancer Lett. 2004;214:19-23 https://doi.org/10.1016/j.canlet.2004.04.012
  9. Buttitta F, Marchetti A, Gadducci A, et al. p53 alterations are predictive of chemoresistance and aggressiveness in ovarian carcinomas: a molecular and immunohistochemical study. Br J Cancer. 1997;75:230-235 https://doi.org/10.1038/bjc.1997.38
  10. Righetti SC, Della Torre G, Pilotti S, et al. A comparative study of p53 gene mutations, protein accumulation, and response to cisplatin-based chemotherapy in advanced ovarian carcinoma. Cancer Res. 1996; 15;56:689-693
  11. Bergh J, Norberg T, Sjogren S, Lindgren A, and Holmberg L. Complete sequencing of the p53 gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy. Nat Med. 1995;1:1029-1034 https://doi.org/10.1038/nm1095-1029
  12. Sanchez Y, Bachant J, Wang H, et al. Control of the DNA damage checkpoint by chk1 and rad53 protein kinases through distinct mechanisms. Science. 1999; 5;286:1166-1171
  13. Wakasugi M, and Sancar A. Order of assembly of human DNA repair excision nuclease. J Biol Chem. 1999;274:18759-18768 https://doi.org/10.1074/jbc.274.26.18759
  14. Batty DP, and Wood RD. Damage recognition in nucleotide excision repair of DNA. Gene. 2000;241:193-204 https://doi.org/10.1016/S0378-1119(99)00489-8
  15. Zou L, and Elledge SJ. Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science. 2003;300:1542-1548 https://doi.org/10.1126/science.1083430
  16. Wang G, Dombkowski A, Chuang L, Xin S, and XU X. The involvement of XPC protein in the cisplatin DNA damaging treatment mediated cellular response. Cell Res. 2004;14:303-314 https://doi.org/10.1038/sj.cr.7290375
  17. Smith WL, DeWitt DL, and Garavito RM, Cyclooxygenases: Structural, cellular, and molecular biology. Annu Rev Biochem. 2000; 69:145–182
  18. Williams CS, Mann M, and DuBois RN, The role of cyclooxygenases in inflammation, cancer, and development. Oncogene. 1999;18:7908–7916
  19. Kim YB, Kim GE, Pyo HR, et al. Differential cyclooxygenase-2 expression in squamous cell carcinoma and adenocarcinoma of the uterine cervix. Clin Invest 2004;60:822-829
  20. Kim SA, Ahn SG, Kim DK, et al. Sequential expression of inducible nitric oxide synthase and cyclooxygenase-2 during DMBA-induced hamster buccal pouch carcinogenesis. In Vivo. 2004;18: 609-614
  21. Keyomarsi K, O'Leary N, Molnar G, Lees E, Fingert HJ, and Pardee AB. Cyclin E, a potential prognostic marker for breast cancer. Cancer Res. 1994;54: 380-385
  22. Tahara E. Genetic alterations in human gastrointestinal cancers. Cancer(suppl.) 1995;75:1410-1417 https://doi.org/10.1002/1097-0142(19950315)75:6+<1410::AID-CNCR2820751504>3.0.CO;2-O
  23. Akama Y, Yasui W, Kuniyasu H, et al. Genetic status and expression of the cyclin-dependent kinase inhibitors in human gastric carcinoma cell lines. Jpn J Cancer Res. 1996;87:824-830 https://doi.org/10.1111/j.1349-7006.1996.tb02106.x
  24. Porter PL, Malone KE, Heagerty PJ, et al. Expression of cell-cycle regulators $p27^{Kip1}$ and cyclin E, alone and in combination, correlate with survival in young breast cancer patients. Nat Med. 1997;3:222-225 https://doi.org/10.1038/nm0297-222
  25. Robbins BA, de la Vega D, Ogata K, Tan EM, and Nakamura RM. Immunohistochemical detection of proliferating cell nuclear antigen in solid human malignancies. Arch Pathol Lab Med. 1987;111:841-845
  26. Hall PA, Levison DA, Woods AL, et al. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990 Dec;162:285-294 https://doi.org/10.1002/path.1711620403
  27. Counter CM, Hirte HW. Bacchetti S, and Harley CB. Telomerase activity in human ovarian carcinoma. Proc Natl acad Sci USA. 1994;91:2900-2904
  28. Counter CM, Gupta J, Harley CB, Leber B, and Bacchetti S. Telomerase activity in normal leukocytes and in hematologic malignancies. Blood. 1995;85:2315-2320
  29. Fujimoro r, Kamata N, Taki M, et al. Gene expression of telomerase related proteins in human normal oral and ectocervical epithelial cells. Oral Oncolo. 2003;39:445-452 https://doi.org/10.1016/S1368-8375(03)00003-4
  30. Culhaci N, Metin K, Copcu E, and Dikicioglu E. Elevated expression of MMP-13 and TIMP-1 in head and neck squamous cell carcinoma may reflect increased tumor invasiveness. BMC Caner. 2004;4:42 https://doi.org/10.1186/1471-2407-4-42
  31. Robinson CM, Stone AM, Shields JD, et al Functional significance of MMP-2 and MMP-9 expression by human malignant oral keratinocyte cell lines. Arch Oral Biol 2003;48:779-786 https://doi.org/10.1016/S0003-9969(03)00172-9
  32. Richard CKJ, Maricris M, Caroline HS, et al. Overexpression of matrix metalloproteinase-1 and -9 mRNA is associated with progression of oral dysplasia to cancer. Clin Cancer Res. 2004;10: 6460-6465 https://doi.org/10.1158/1078-0432.CCR-04-0656
  33. Birkedal-Hansen B, Pavelic ZP, Gluckman JL, Stambrook P, Li YQ, and Stetler-Stevenson WG. MMP and TIMP gene expression in head and neck squamous cell carcinomas and adjacent tissues. Oral Dis. 2000 ;6:376-382 https://doi.org/10.1111/j.1601-0825.2000.tb00130.x