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研究生:林庭蕙
研究生(外文):TING-HUI,LIN
論文名稱:維生素C抑制顆粒蛋白誘發腦癌細胞的增生
論文名稱(外文):Vitamin C inhibit granulin-induced proliferation on Brain cancer cells
指導教授:魏秋偉魏秋偉引用關係
指導教授(外文):CHYOU-WEI,WEI
學位類別:碩士
校院名稱:弘光科技大學
系所名稱:營養醫學研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
畢業學年度:99
語文別:中文
論文頁數:52
中文關鍵詞:顆粒蛋白前體腦癌細胞增殖維生素C凋亡
外文關鍵詞:particle precursor proteinbrain cancercell proliferationvitamin Capoptosis
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Progranulin (Pgrn)是一種醣蛋白質,它可以調整細胞週期,增殖和傷口癒合。許多研究報告顯示Pgrn過度增生可見於癌症病患身上,例如乳癌、卵巢癌、前列腺癌和肝癌。在過去探討Granulin誘發腫瘤增生的研究都用在人體或動物,未使用在細胞研究上。
本研究模擬建立人體微環境探討Granulin誘發人類惡性腦癌細胞(DBTRG細胞)的增生;而維生素C為一有效之抗氧化物,過去研究曾被用於伴隨治療Pgrn所導致的腫瘤擴散。
本實驗即利用不同濃度的維生素C加入含Granulin的腦癌細胞,看維生素C是否能抑制Granulin造成的腫瘤細胞增生。初步的結果顯示,grn藉由dose-dependent的方式,可以誘導DBTRG細胞的擴散。維生素C可抑制grn所誘導的DBTRG 細胞增殖,且使細胞走向凋亡。因此在未來應用於癌細胞的治療上,具有一參考價值。


關鍵字: 顆粒蛋白前體、腦癌、細胞增殖、維生素C、凋亡

Progranulin (Pgrn)(also called granulin or epithelin precursor, axrogranin or PC-derived growth factor) is a glycoprotein, which is used to adjust the cell cycle, proliferation, and wound healing. Many empirical studies or experiments show that progranulin contributes to tumorigenesis in diverse cancers, including breast cancer, ovarian cancer, prostate cancer and liver cancer. Most of previous studies or researches in the field are focus on human or animal, but only few on the cell.It is widely recognized that vitamin C is one of the effective antioxidants and has been used to cure the human tumors resulted from the overuse of Pgrn.

This study attempts to build a simulation model in a human-induced micro-environment, examining how vitamin C inhibits the progranulin induction in the DBTRB cell.This study attempts to add various dozen of Vitamin C in the DBTRG cell with the progranulin, observing if vitamin C would be able to curb the continuous growth of such evil cells.

The preliminary data given by this study reveals that the Vitamin C can inhibit the proliferation of DGTRG cells induced by the Pgrn through the dose-dependant. Furthermore, the vitamin C ends up pushing those cells toward apoptosis.Thus,this study has provided some useful implications for the researches interested in the treatment of cancer cells.




Keywords: particle precursor protein, brain cancer, cell proliferation, vitamin C,
apoptosis

中文摘要……………………………………………………………………….....2
英文摘要………………………………………………………………………….3
目 錄………………………………………………………………………….4
謝 誌………………………………………………………………………….6
縮 寫 表………………………………………………………………………….7
第一章 緒 論………………………………………………………………….8
1 研究動機………………………………………………………………….9
2 研究目的....................................................................................................9
3 研究步驟…………………………………………………………………10
4 研究架構…………………………………………………………………11
第二章 文獻回顧………………………………………………………………..12
1 維生素C(Vitamin C)...............................................................................12
2 顆粒蛋白前體( Progranulin )..................................................................17
3 腦癌細胞(Brain Cancer Cells )...............................................................18
4 細胞週期(Cell Cycle)和細胞凋亡(Apoptosis)........................................19
5 自由基(Free Radical)...............................................................................20
6 抗氧化酵素系統.......................................................................................21
第三章 實驗方法與材料
1 細胞培養…………………………………………………………………27
2 細胞存活率測定......................................................................................27
3 細胞自由基………………………………………………………………28
4 統計分析…………………………………………………………………28
5 實驗流程…………………………………………………………………28
第四章 結果……………………………………………………………………..31
1 實驗ㄧ 不同濃度的人類、魚類grn作用於DBTRG細胞後生長情
形及自由基含量之變化…………………………………………………31
1-1最適生長濃度及天數………………………………………………….31
1-2 不同濃度人類grn處理DBTRG 細胞後對細胞存活率之影響
…………………………………………………………………………31
1-3 不同濃度人類grn處理DBTRG細胞後對細胞自由基含量變化
…………………………………………………………………………31
1-4 不同濃度魚類grn處理DBTRG 細胞後,對細胞存活率之影響
………………………………………………………………………..31
1-5 不同濃度魚類grn處理DBTRG細胞後,對細胞自由基含量變化
………………………………………………………………………..32
1-6 不同濃度人類grn於正常腦細胞mRTP2A後,對細胞存活率之
影響.....................................................................................................32
1-7 不同濃度魚類grn於正常腦細胞mRTP2A後,對細胞存活率之
影……………………………………………………………………..32
2 實驗二 人類、魚類grn作用於DBTRG細胞後加入維生素C觀察
細胞生長情形及是否造成細胞凋………………………………….33
2-1 人類grn處理DBTRG 細胞後加入維生素C 5M對細胞生長情
形之影響…………………………………………………………….33
2-2 魚類grn處理DBTRG 細胞後加入維生素C 5M對細胞生長情
形之影響…………………………………………………………….33
2-3 維生素C處理加入人類及魚類grn的DBTRG細胞造成細胞
凋亡或壞死………………………………………………………….33
第五章 討論與建議……………………………………………………………39
1 人類grn處理DBTRG細胞後對細胞存活率影響之探討…………. 39
2 人類grn處理DBTRG細胞後對細胞自由基含量變化之影響……. 39
3 魚類grn處理DBTRG細胞後對細胞存活率之探討………………. 40
4 比較魚類和人類的grn處理DBTRG細胞後對細胞自由基含
量變化之探討…………………………………………………………..40
5 探討人類和魚類grn對正常腦細胞mRTP2A及腦瘤細胞
存活率之影響…………………………………………………………..40
6 抗壞血酸抑制腫瘤細胞之探討……………………………………….41
7 探討維生素C造成腫瘤細胞發生細胞凋亡…………………………..41
8 結論與建議…………………………………………………………….42
表目錄………………………………………………………………………….43
圖目錄………………………………………………………………………….43
參考文獻……………………………………………………………………….44

參 考 文 獻
1. Ardelt W, Mikulski SM, Shogen K. (1991) Amino acid sequence of an anti-tumor protein from Rana pipiens oocytes and early embryos. Homology to pancreatic ribonucleases. J Biol Chem. 266(1):245-51.

2. Arnold U, Ulbrich-Hofmann R. (2006) Natural and engineered ribonucleases as potential cancer therapeutics. Biotechnol Lett. 28(20):1615-1622

3. Barbero S, Bonavia R, Bajetto A, Porcile C, Pirani P, Ravetti JL, Zona GL, Spaziante R, Florio T, Schettini G. (2003) Stromal cell-derived factor 1alpha stimulates human glioblastoma cell growth through the activation of both extracellular signal-regulated kinases 1/2 and Akt. Cancer Res. 63(8):1969-74.

4. Beg AA, Baltimore D. (1996) An essential role for NF-kappa B in preventing TNF-α-induced cell death. Science. 274: 782-784.

5. Benito A, Ribo M,Vilanova M. (2005) On the track of antitumor ribonuclease. Mol. BioSyst. 1(4):294-302

6. Chang CF, Chen C, Chen YC, Hom K, Huang RF, Huang TH. (1998) The solution structure of cytotoxic ribonuclease from the oocytes of Rana catebeiana (bullfrog). J Mol Biol. 283(1): 231-244.

7. Cheng EH, Levine LH, Boise CB, Hardwick JM. (1996) Bax-independent inhibition of apoptosis by Bcl-XL. Nature. 379: 554-556.

8. Cheng EH, Kirsch RJ, Ravi MB, Bedi K, Hardwick JM. (1997) Conversion of Bcl-2 to a Bax-like death effector by caspases. Science. 278: 1966-1968.

9. Clem RJ, Cheng EH, Karp CL, Karp DG, Kirsch K, Takahashi A, Kastan MB, Griffin DE, Earnshaw WC, Veliuona MA, Hardwich JM. (1998) Modulation of cell death by Bcl-XL through caspase interaction. Proc. Natl. Avad. Sci.USA 95:554-559.

10. Coste A, Farbre M, McDonell N, Porteu A, Gilgenkrantz H, Perret C, Kahn A, Mignon A. (1999) Differential protective effects of Bcl-XL and Bcl-2 on apoptotic liver injuryin transgenic mice. Am. J. Physiol. 277: G702-G708.


11. Darnbrough C, Ford PJ. (1979) Turnover and processing of poly(A) in full-grown oocytes and during progesterone-induced oocyte maturation in Xenopus laevis. Dev Biol. 71(2): 323-340.

12. Domachowske JB, Dyer KD, Adams AG, Leto TL, Rosenberg HF. (1998a) Eosinophil cationic protein/Rnase 3 is another RNase A-family ribonuclease with direct antiviral activity. Nucleic Acid Res. 26(14): 3358-3363.

13. Domachowske JB, Dyer KD, Bonville CA, Rosenberg HF. (1998b) Recombinant human eosinophil-derived neurotoxin/RNase 2 functions as an effective antiviral agent against respiratory syncytial virus. J Infect Dis. 177(6): 1458-1464.

14. Domachowske JB, Bonville CA, Dyer KD, Rosenberg HF. (1998c) Evolution of antiviral activity in the ribonuclease A gene superfamily: evidence for a specific interaction between eosinophil-derived neurotoxin (EDN/RNase2) and respiratory syncytial virus. Nucleic Acid Res. 26(23): 5327-5332.

15. Dong L, Wang W, Wang F, Stoner M, Reed JC, Harigai M, Samudio I, Kladde MP, Vyhlidal C, Safe S. (1999) Mechanisms of transcriptional activation of bcl-2 gene expression by 17beta-estradiol in breast cancer cells.J Biol Chem. 274(45): 32099-107.

16. Elstner E, Williamson EA, Zang C, Fritz J, Heber D, Fenner M, Possinger K, Koeffler HP. (2002) Novel therapeutic approach: ligands for PPARγ and retinoid receptors induce apoptosis in bcl-2-positive human breast cancer cells. Breast Cancer Res and Treat 74:155-165.

17. Gottardis M, Jordan C. (1988) Development of tamoxifen-stimulated growth of MCF-7 tumors in athymic mice after long-term antiestrogen administration. Cancer Research. 48: 5183-5187.

18. Hsu CK, Risch A, Li XS, Dawson M, Reichert U, Shroot B, Fontana J. (1997) Bcl-XL expression and its downregulation by a novel retinod inbreast carcinoma cells. Exprimental Cell Res. 232: 17-24.

19. Huang HC, Wang SC, Leu YJ, Lu SC, Liao YD. (1998) The Rana catesbeiana rcr gene encoding a cytotoxic ribonuclease. Tissue distribution, cloning, purification, cytotoxicity, and active residues for RNase activity. J Biol Chem. 273(11): 6395-6401.

20. Hu CC, Lee YH, Tang CH, Cheng JT, Wang JJ. (2001a) Synergistic cytotoxicity of Rana catebeiana ribonuclease and IFN-gamma on hepatoma cells. Biochem Biophys Res Commun. 280(5): 1229-1236.

21. Hu CC, Tang CH, Wang JJ. (2001b) Caspase activation in response to cytotoxic Rana catesbeiana ribonuclease in MCF-7 cells. FEBS Lett. 503(1):65-8.

22. Kawauchi H, Sakakibara F, Watanabe K. (1975) Agglutinins of frog eggs: anew class of proteins causing preferential agglutination of tumor cells. Experientia. 31(3): 364-365.
23.Lheureux S, Le Moulec S.(2011) Molecules involved in apoptosis regulation as therapeutic targets in oncology. Bull Cancer. 24(6);452-458.
24. Reimann M, Schmitt CA, Lee S.(2011) Non-cell-autonomous tumor suppression: oncogene-provoked apoptosis promotes tumor cell senescence via stromal crosstalk.J Mol Med (Berl). 19(6);267-272.

25. Lee YH, Wang JJ (2001) The effects of Rana catebeiana RNase on Japanese encephalitis virus replication in BHK-21 cells. Mol Biol Cell 12:249a. The 41st Annual Metting of American Society for Cell Biology. Nov. 8-12, 2001. Washington DC, USA.

26. Leland PA, Raines RT. (2001) Cancer chemotherapy ribonucleases to the rescue. Chem Biol. 8(5): 405-13. Review.

27. Leung K, Wang TY. (1999) Differential ffevts of chemotherapeutic agents on Bcl-2/Bax apoptosis pathway inhuman breast cancer cell line MCF-7. Breast Cancer Res and Treat. 55: 73-83.

28. Liao YD. (1992) A pyrimidine-guanine sequence-specific ribonuclease from Rana catesbeiana (bullfrog) oocytes. Nucleic Acids Res. 20:1371-1377.

29. Liao YD, Wang JJ. (1995) Yolk granules are the major compartment for bullfrog (Rana catebeiana) oocyte-specific ribonuclease. Eur J Biochem. 222(1): 213-220.

30. Liao YD, Huang HC, Chen HJ, Kuo SJ. (1996) Large-scale preparation of a ribonuclease from Rana catebeiana (bullfrog) oocytes and characterization of its specific cytotoxic activity against tumor cells. Protein Expression Purif. 7: 194-202.

31. Liao YD, Huang HC, Leu YJ, Wei CW, Tang PC, Wang SC. (2000) Purification and cloning of cytotoxic ribonucleases from Rana catebeiana (bullfrog). Nucleic Acids Res. 28(21): 4097-4104.

32. Liu G, Svhwartz JA, Brooks SC. (1999) p53 down-regulates ER-Responsive gene by interfering with the binding of ER to ERE. Biochem. Biophy. Res. Commun. 264: 359-364.

33. Michaelis M, Cinatl J, Anand P, Rothweiler F, Kotchetkov R, Deimling A, Doerr HW, Shogen K, Cinatl J. (2007) Onconase induces caspase-independent cell death in chemoresistant neuroblastoma cells. Cancer Lett. 250(1):107-16.

34. Newport J, Kirschner M. (1982) A major developmental transition in early Xenopus embryos: II, Control of the onset of transcription. Cell. 30(3): 687-696.

35. Newton DL, Nicholls PJ, Rybak SM, Youle RJ. (1994) Expression and characterization of recombinant human eosinophil-derived neurotoxin and eosinophil-derived neurotoxin-anti-transferrin receptor sFv. J Biol Chem. 269(43): 26739-45.

36. Newton DL, Rybak SM. (1998) Unique recombinant human ribonuclease and inhibition of Kaposi’s sarcoma cell growth. J Natl Cancer Inst. 90(23): 1787-91.

37. Nitta K, Ozaki K, Ishikawa M, Furusawa S, Hosono M, Kawauchi H, Sasaki K, Takayanagi Y, Tsuiki S, Hakomori S. (1994) Inhibition of cell proliferation by Rana catebeiana and Rana japonica lectins belonging to the ribonuclease superfamily. Cancer Res. 54(4): 920-927.

38. Obrero M, Yu D, Shapiro J. (2002) Estrogen receptor-dependent and estrogen receptor-independent pathways for tamoxifen and 4-hydroxytamoxifen-induced programmed cell death. J. Biol. Chem. 277(47): 45695-45703.

39. Pao CI, Lee TC, Liao YD, Wu CW. (1988) An N-terminally fused Xenopus transcription factor IIIA synthesized in Escherichia coli is biologically active. J Biol Chem. 263(21): 10295-10299.

40. Progranulin gene expression regulates epithelial cell growth and promotes tumor growth in vivo: Z. He & A. Bateman; Cancer Res. 59, 3222 (1999)
41. Progranulin: normal function and role in neurodegeneration: J.L. Eriksen & I.R. Mackenzie; J. Neurochem. 104, 287 (2008)
42.Progranulin is a mediator of the wound response: Z. He, et al.; Nat. Med. 9, 225 (2003)

43. Real P, Sierra A, Juan A, Segovia J, Lopez-Vega J, Fernandez-Luna J. (2002) Resistance to chemotherapy via Stat3-dependent overexpression of Bcl-2 in metastatic breast cancer cells. Oncogene. 21: 7611-7618.

44. Rosenberg H, Dyer K. (1996) Molecular cloning and characterization of a novel human ribonuclease (RNAse k6): increasing diversity in the enlarging ribonuclease gene family. Nucleic acid Res. 18: 3507-3513.

45. Saxena SK, Gravell M, Wu YN, Mikulski SM, Shogen K, Ardelt W, Youle RJ. (1996) Inhibition of HIV-1 production and selective degradation of viral RNA by an amphibian ribonuclease. J Biol Chem. 271(34): 20783-10788.

46. Schein CH. (1997) From housekeeper to microsurgeon: the diagnostic and therapeutic potential of ribonucleases. Nat Biotechnol. 15(6): 529-536.

47. Shapiro R, Riordan JF, Vallee BL. (1986) Characteristic ribonucleolytic activity of human angiogenin. Biochemistry. 25(12): 3527-3532.

48. Simonian L, Grillot AM, NuNez G. (1997a) Bcl-2 and Bcl-XL can differentially block chemotherapy-induced cell death. Blood. 90(3): 1208-1216.

49. Simonian L, Grillot AM, NuNez G. (1997b) Bak can accelerate chemotherapy-induced cell death independently of its heterodimerization with Bcl-XL and Bcl-2. Oncogene. 15: 1871-1875.

50. Tao R, Yamane H, Mori H, Gradziel TM, Dandekar AM, Sugiura A. (1997) Identification of stylar Rnases associated with gametophytic self-incompatibility in almomd (Pruns dulcis). Plant Cell Physiol. 38(3): 304-311.

51. Titani K, Takio K, Kuwada M, Nitta K, Sakakibara F, Kawauchi H, Takayanagi G, Hakomori S. (1987) Amino acid sequence of sialic acid binding lectin from frog (Rana catebeiana) eggs. Biochemistry. 26(8): 2189-2194.

52. Tsai NM, Chen YL, Lee CC, Lin PC, Cheng YL, Chang WL, Lin SZ, Harn HJ. (2006) The natural compound n-butylidenephthalide derived from Angelica sinensis inhibits malignant brain tumor growth in vitro and in vivo. J Neurochem. 99(4):1251-62.
53. Wang JJ, Tang PC, Chao SH, Cheng CH, Ma HJ, Liao YD. (1995) Immunocytochemical localization of ribonuclease in yolk granules of adult Rana catebeiana oocytes. Cell Tissue Res. 280(2): 259-265.

54. Wei CW, Hu CC, Tang CH, Lee MC, Wang JJ. (2002) Induction of differentiation resues HL-60 cells from Rana catebeiana ribonuclease-induced cell death. FEBS Letters. 531: 421-426.

55. Guo Y, Kyprianou N. (1999) Restoration of transforming growth factor beta signaling pathway in human prostate cancer cells suppresses tumorigenicity via induction of caspase-1-mediated apoptosis. Cancer Res. 59(6):1366-71.

56. Youle RJ, Wu YN, Mikulski SM, Shogen K, Hamilton RS, Newton D, D’Alessio G, Gravell M. (1994) RNase inhibition of human immunodeficiency virus infection of H9 cells. Proc. Natl Adad Sci U.S.A 91(13): 6012-6.

57. Yang E, Korsmeyer S. (1996) Molecular thanatopsis: a discourse on the Bcl-2 family and cell death. Blood. 88(2): 386-401.

58. Yin D, Zhou H, Kumagai T, Liu G, Ong JM, Black KL, Koeffler HP. (2005) Proteasome inhibitor PS-341 causes cell growth arrest and apoptosis in human glioblastoma multiforme (GBM). Oncogene. 24(3):344-54.

59. Youle RJ, Wu YN, Mikulski SM, Shogen K, Hamilton RS, Newton D, D’ Alessio G, Chen c, Hom K, Huang RF, Chong PJ, Liao YD, Huang TH. (1996) Secondary structure of a pyrimidine-guanine sequence specific ribonuclease possessing cytotoxic activity from the oocytes of Rana catebeiana. J Biomol. NMR. 8: 331-334.

60. Zhang GJ, Kimijima I, Onda M, Kanno M, Sato H, Watanabe T, Tsuchiya A, Rikiya A, Takenoshita S. (1999) Tamoxifen-induced apoptosis in breast cancer cells relates to down-regulation of bcl-2, but not bax and bcl-XL, without alteration of p53 protein levels. Clinical Cancer Res. 5: 2971-2977.
61. Zou A, Marschke KB, Arnold KE, Berger EM, Fitzgerald P, Mais DE, Allegretto EA. (1999) Estrogen receptor beta activates the human retinoic acid receptor alpha-1 promoter in response to tamoxifen and other estrogen receptor antagonists, but not in response to estrogen. Mol Endocrinol. 13(3): 418-30

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