Journal of Pharmacopuncture (대한약침학회지)

KOREAN PHARMACOPUNCTURE INSTITUTE (대한약침학회)
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DNA and Proteomic Expression of Cervi parvum cornu Herbal-acupuncture Solution (CPC-HAS) in HepG2 carcinomar cells

녹용약침액(鹿茸藥鍼液)의 간암세포주(肝癌細胞柱)에 대한 DNA 및 단백질 발현(發顯)

  • Ryu, Sung-Hyun (Department of Acupuncture & Moxibustion, College of Oriental Medicine, Daegu Haany University) ;
  • Lee, Kyung-Min (Department of Acupuncture & Moxibustion, College of Oriental Medicine, Daegu Haany University) ;
  • Lee, Bong-Hyo (Department of Acupuncture & Moxibustion, College of Oriental Medicine, Daegu Haany University) ;
  • Lim, Seong-Chul (Department of Acupuncture & Moxibustion, College of Oriental Medicine, Daegu Haany University) ;
  • Jung, Tae-Young (Department of Acupuncture & Moxibustion, College of Oriental Medicine, Daegu Haany University) ;
  • Seo, Jung-Chul (Department of Acupuncture & Moxibustion, College of Oriental Medicine, Daegu Haany University)
  • 류성현 (대구한의대학교 한의과대학 침구경혈학교실) ;
  • 이경민 (대구한의대학교 한의과대학 침구경혈학교실) ;
  • 이봉효 (대구한의대학교 한의과대학 침구경혈학교실) ;
  • 임성철 (대구한의대학교 한의과대학 침구경혈학교실) ;
  • 정태영 (대구한의대학교 한의과대학 침구경혈학교실) ;
  • 서정철 (대구한의대학교 한의과대학 침구경혈학교실)
  • Published : 2006.06.30
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Abstract

Objective : It has long been known about the osteogenic effect of CPC-HAS on bone tissues. However, it has not been determined the effect of CPC-HAS on cancer cells. The purpose of this study is to screen the CPC-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells. Oligonucleotide microarray and proteomics approaches were employed to screen the differential expression genes. Methods : CPC-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CPC-HAS (0.1, 0.5, 1.5, 10, 20mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cell in all concentrations(0.l, 0.5, 1.5, 10, 20mg/ml). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 23 with 5 up-regulated and 18 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. Two down-regulated proteins were aldehyde dehydrogenase 1 and enolase 1, and up-regulated protein was fatty acid binding protein 1 by 1.5mg/ml of CPC-HAS. Discussion : This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray and proteomic analysis. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.

Keywords

References

  1. 서울대학교 의과대학. 종양학. 서울: 서울대학교 출판부. 1992 : 1, 3, 27, 36, 91, 99, 137-192.
  2. 통계청. 2003년 사망원인통계연보. 대전: 통계청. 2004 : 22-4.
  3. 박재갑. 인간생명과학. 서울: 서울대학교출판부. 1993:662-73.
  4. El-Aneed A. Current strategies in cancer gene therapy. European Journal of Pharmacology. 2004;498(1-3):1–18. https://doi.org/10.1016/j.ejphar.2004.06.054
  5. Guimaraes MJ, Lee F, Zlotnik A, McClanahan T. Differentia1 display by PCR:novel findings and applications. Nucleic Acids Res 1995;23(10):1832–3. https://doi.org/10.1093/nar/23.10.1832
  6. Kurian K, Watson CJ, Wyllie AH. DNA chip technology. J Pathos 1999;187(3):267–71.
  7. DeRisi JL, Penland L, Brovn PO. Use of a cDNA microarray to analyse gene expression patterns in human cancer. Nature Genet 1996;14(4):457–60. https://doi.org/10.1038/ng1296-457
  8. Sapolsky RJ, Lipshultz RJ. Mapping genomic library clones using oligonucleotide arrays. Genomics 1996;33(3):445–56. https://doi.org/10.1006/geno.1996.0219
  9. Collins FS, Patrinos A, Jordan E, Chakravarti A, Gesteland R, Walters L. New goals for the U.S. human genome project: 1998-2003. Science 1998;282(5389):682–746. https://doi.org/10.1126/science.282.5389.682
  10. 김정숙. 생약을 이용한 골다공증 치료제 개발연구. 제 1회 골다공증 연구회 심포지움 proceeding 1998;1:32–6.
  11. 강성길, 김대수. 녹용, 인삼 및 영지수침이 면역반응에 미치는 영향. 경희의학 1988;8(2):138–54.
  12. 김남재, 심상범, 김재근, 원도희, 홍남두. 녹용 물 추출물과 항암제의 병용투여에 관한 연구. 생약학회지 1998;29(2):93–104.
  13. 中國醫藥院. 申農本草經. 集文書局. 臺北. 1976 : 27.
  14. 한상원, 서정철, 이윤호, 최제용. 鹿茸藥鍼液의 DNA chip을 利用한 遺傳子 發顯 分析. 大韓鍼灸學會誌 2003;20(3):34–44.
  15. 楊維傑. 編. 黃帝內經素門靈樞錫解. 성보사. 서울. 1980: 41, 45, 97, 168, 243, 295-296, 407, 447, 469, 473, 577.
  16. 許俊. 東醫寶鑑. 서울: 南山堂. 1988 : 486-96, 720.
  17. 賈堃. 癌瘤防治硏究. 서울: 成輔社. 1984: 25-7.
  18. 全國韓醫科大學 鍼灸.經穴學敎室 編. 鍼灸學<下>. 4. 서울 : 集文堂. 1994: 1457-67
  19. 임사비나, 한상원, 변부형. 人蔘藥鍼과 Lidocaine을 添加한 人蔘藥鍼이 腫瘍 및 免疫機能에 미치는 影響. 東西醫學 1995;20(3):21–40.
  20. 김소형, 김갑성. 全蝎 藥鍼液의 抗突然變異 및 抗癌 效果. 大韓鍼灸學會誌 2000;17(3):151–67.
  21. 全國韓醫科大學 本草學敎室 編. 本草學. 서울: 永林社. 1991 : 545-6.
  22. 보건사회부. 대한약전 외 한약(생약)규격집. 서울: 보건사회부. 1987:98.
  23. 한상원, 김영태, 손양선, 진수희, 심인섭, 임사비나 외. 鹿茸 및 鹿茸藥鍼이 動物의 成長과 知能發達에 미치는 影響. 大韓鍼灸學會誌 2001;18(5):122–35.
  24. 李敏炯, 徐榮培. 鹿茸의 造血作用에 對한 實驗的 硏究. 大韓本草學會誌 2001;16(1):91–110.
  25. 용재익. Cholesterol 投與 家兎의 血淸 中 cholesterol에 미치는 鹿茸의 影響. 약제학회지 1976;6(3):314–20.
  26. Schuler GD, Bonguski MS, Stewart EA, Stein LP, Gyapay G, Rice K, et al. A gene map of the human genome. Science 1996;274(5287):540–6. https://doi.org/10.1126/science.274.5287.540
  27. Hedric SM, Cohen DI, Nielsen EA, Davis MM. Isolation of cDNA clones encoding T-cell specific membrane associated proteins. Nature 1984;308(5955):149–53. https://doi.org/10.1038/308149a0
  28. Liang P, Pardee AB. Differential disp1ay of eukaryotic messenger RNA by means of polymerase chain reaction. Science 1992;257(5072):967–71. https://doi.org/10.1126/science.1354393
  29. Mason IJ, Taylor A, Williams JG, Sage H, Hugan BLM. Evidence from molecular cloning that SPARC, a major product of mouse embryropariental endoderm is related to an endothelial cell cu1ture shock glycoprotein of Mr. 43000. EMBO J 1986;5(9):1465–72.
  30. Liang P, Averboukh L, Keyomarsi K, Sager R, Pardee AB. Differential display and cloning of messenger RNAs from human breast cancer versus mammary epithelial cells. Cancer Res. 1992;52(24):6966–8.
  31. http://www.affymetrix.com.
  32. Wada T, Penninger JM. Mitogen-activated protein kinases in apoptosis regulation. Oncogene 2004;23(16):2838–49. https://doi.org/10.1038/sj.onc.1207556
  33. Han C, Demetris AJ, Liu Y, Shelhamer JH, Wu T. Transforming growth factor-beta (TGF-beta) activates cytosolic phospholipase A2alpha (cPLA2alpha)-mediated prostaglandin E2 (PGE)2/EP1 and peroxisomeproliferator -activated receptor-gamma (PPAR-gamma)/Smad signaling pathways in human liver cancer cells. A novel mechanism for subversion of TGF-beta-induced mitoinhibition. J Biol Chem 2004;279(43):44344–54. https://doi.org/10.1074/jbc.M404852200
  34. Shpargel KB, Ospina JK, Tucker KE, Matera AG, Hebert MD. Control of Cajal body number is mediated by the coilin C-terminus. J Cell Sci 2003;116(Pt 2):303–12. https://doi.org/10.1242/jcs.00211
  35. Sun QA, Wu Y, Zappacosta F, Jeang KT, Lee BJ, Hatfield DL, Gladyshev VN. Redox regulation of cell signaling by selenocysteine in mammalian thioredoxin reductases. J Biol Chem 1999;274(35):24522–30. https://doi.org/10.1074/jbc.274.35.24522
  36. Ko J, Ryu KS, Lee YH, Na DS, Kim YS, Oh YM, Kim IS, Kim JW. Human secreted frizzled-related protein is down-regulated and induces apoptosis in human cervical cancer. Exp Cell Res 2002;280(2):280–7 https://doi.org/10.1006/excr.2002.5649
  37. Spence JP, Liang T, Eriksson CJ, Taylor RE, Wall TL, Ehlers CL, Carr LG. Evaluation of aldehyde dehydrogenase 1 promoter polymorphisms identified in human populations. Alcohol Clin Exp Res 2003;27(9):1389–94 https://doi.org/10.1097/01.ALC.0000087086.50089.59
  38. Ludwig A, Dietel M, Lage H. Identification of differentially expressed genes in classical and atypical multidrug-resistant gastric carcinoma cells. Anticancer Res 2002;22(6A):3213–21.