研究生: |
沈志謙 Chih-Chien, Shen |
---|---|
論文名稱: |
探討五種中藥單方處理非小細胞肺癌細胞株之效果 Investigation of the effect of five Chinese herbs in non-small cell lung carcinoma cell lines |
指導教授: |
謝秀梅
Hsieh, Hsiu-Mei |
學位類別: |
碩士 Master |
系所名稱: |
生命科學系 Department of Life Science |
論文出版年: | 2011 |
畢業學年度: | 99 |
語文別: | 中文 |
論文頁數: | 79 |
中文關鍵詞: | 非小細胞肺癌 、中草藥 、癌幹細胞 |
英文關鍵詞: | NSCLC, Chinese herbs, CSCs |
論文種類: | 學術論文 |
相關次數: | 點閱:208 下載:6 |
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替代性療法近來在許多疾病中因為療效良好而備受關注;其好處
為除了可以有效的治療疾病之外,相較之下,其所帶來的毒性或是副
作用可能低於傳統的治療方式。多年以來,肺癌即是許多國家癌症死
亡的首位,而本研究主要的目的就是想要探討傳統的中草藥配方是否
對於非小細胞肺癌細胞株A549, NCI-H460, NCI-H520 具有抑制其生
長或是存活的效果。我們對細胞分別用了五種不同的中草藥
(HC1-HC5)作處理,再探討這些藥物是透過甚麼樣的機制去減低細胞
生長的情形。我們發現在五種中草藥方內有兩種中草藥HC3以及HC4
對於減低非小細胞肺癌細胞株之存活率有顯著的影響,但相同的劑量
對於正常細胞株MRC5 則沒有太大的影響。另外我們也以流式細胞
儀以及西方墨點轉漬的實驗進一步的探討藥物處理對於細胞週期的
表現影響,我們發現在藥物處理後細胞族群較多停滯在G1 以及G2
的現象,分析調節細胞週期的蛋白質表現結果也符合流式細胞儀看到
的結果。除此之外,我們也發現了A549 側群細胞(具有類癌幹細胞的
特性)的比例在經過藥物處理後有顯著降低的趨勢,我們也分析了間
質-表皮轉換(MET)的相關蛋白以進一步的解釋癌幹細胞可能的變化
機制。另外我們也觀測到癌細胞的轉移能力因為藥物的處理而降低。
由我們的分析結果顯示這些藥物也許有潛力可以應用於肺癌治療。
Complementary Alternative Medicines (CAM) is attractive for its
therapeutic efficacy, and its less toxic effect in some diseases than
conventional treatments. Lung cancer has been the leading cause of
cancer death in many countries since a few years ago. In this study we
investigate if some of the Chinese Herbs have the therapeutic potential to
suppress the proliferation or survival rate of Non-Small Cell Lung
carcinoma (NSCLC) cell lines- A549, NCI-H460, and NCI-H520. We
used five different herbal complexes (HC1-5) to treat the cells, and
further investigated the mechanism that contribute to the reduction of cell
survival rate. Two of the five complexes, HC3 and HC4, reduced the
viability of cancer cells but not normal cells under the half maximal
inhibitory concentration (IC50) dosage. The results of flow cytometry
analysis indicated that these two HCs induced cell cycle arrested at G1
and G2 phases. We also confirmed this result by western blot analysis
showing that the expression levels of several cell-cycle regulators,
CDK4/6, cyclin D3 and cyclin B1were reduced after HC3 and HC4
treatment. In addition, side population (SP, which represented stem-like
cancer cells) was also reduced after the treatment. To further elucidate the
molecular mechanism, we examined the mesenchymal to epithelial
transition (MET) on cells after treatment. Furthermore, we also
performed the transwell assay to elucidate the cell migration ability after
treatment. The data showed that both of HCs decreased the migration
capacity of A549. In conclusion, these HCs may have potential to be
applied to the NSCLC therapy.
衛生署。(2010)。中華民國國人98年度十大死因。
王天駿。(2010)。以A549肺癌細胞株為模式篩選對非小細胞肺癌有治療潛力之中藥複方。國立臺灣師範大學研究所碩士論文。
Acloque H, Adams MS, Fishwick K, Bronner-Fraser M, Nieto MA (2009) Epithelial-mesenchymal transitions: the importance of changing cell state in development and disease. J Clin Invest 119:1438-1449.
Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A 100:3983-3988.
Biesalski HK, Bueno de Mesquita B, Chesson A, Chytil F, Grimble R, Hermus RJ, Kohrle J, Lotan R, Norpoth K, Pastorino U, Thurnham D (1998) European Consensus Statement on Lung Cancer: risk factors and prevention. Lung Cancer Panel. CA Cancer J Clin 48:167-176; discussion 164-166.
Blagosklonny MV, Pardee AB (2002) The restriction point of the cell cycle. Cell Cycle 1:103-110.
Catelinois O, Rogel A, Laurier D, Billon S, Hemon D, Verger P, Tirmarche M (2006) Lung cancer attributable to indoor radon exposure in france: impact of the risk models and uncertainty analysis. Environ Health Perspect 114:1361-1366.
Chen LG, Hung LY, Tsai KW, Pan YS, Tsai YD, Li YZ, Liu YW (2008) Wogonin, a bioactive flavonoid in herbal tea, inhibits inflammatory cyclooxygenase-2 gene expression in human lung epithelial cancer cells. Mol Nutr Food Res 52:1349-1357.
Chen Q, Wang Y, Xu K, Lu G, Ying Z, Wu L, Zhan J, Fang R, Wu Y, Zhou J (2010) Curcumin induces apoptosis in human lung adenocarcinoma A549 cells through a reactive oxygen species-dependent mitochondrial signaling pathway. Oncol Rep 23:397-403.
Cheng YW, Chiou HL, Sheu GT, Hsieh LL, Chen JT, Chen CY, Su JM, Lee H (2001) The association of human papillomavirus 16/18 infection with lung cancer among nonsmoking Taiwanese women. Cancer Res 61:2799-2803.
Chiba T, Kita K, Zheng YW, Yokosuka O, Saisho H, Iwama A, Nakauchi H, Taniguchi H (2006) Side population purified from hepatocellular carcinoma cells harbors cancer stem cell-like properties. Hepatology 44:240-251.
Chiu HF, Cheng MH, Tsai SS, Wu TN, Kuo HW, Yang CY (2006) Outdoor air pollution and female lung cancer in Taiwan. Inhal Toxicol 18:1025-1031.
Coyle YM, Minahjuddin AT, Hynan LS, Minna JD (2006) An ecological study of the association of metal air pollutants with lung cancer incidence in Texas. J Thorac Oncol 1:654-661.
Donnenberg VS, Landreneau RJ, Donnenberg AD (2007) Tumorigenic stem and progenitor cells: implications for the therapeutic index of anti-cancer agents. J Control Release 122:385-391.
Faheina-Martins GV, da Silveira AL, Ramos MV, Marques-Santos LF, Araujo DA (2011) Influence of fetal bovine serum on cytotoxic and genotoxic effects of lectins in MCF-7 cells. J Biochem Mol Toxicol.
Fang D, Nguyen TK, Leishear K, Finko R, Kulp AN, Hotz S, Van Belle PA, Xu X, Elder DE, Herlyn M (2005) A tumorigenic subpopulation with stem cell properties in melanomas. Cancer Res 65:9328-9337.
Giuliani L, Jaxmar T, Casadio C, Gariglio M, Manna A, D'Antonio D, Syrjanen K, Favalli C, Ciotti M (2007) Detection of oncogenic viruses SV40, BKV, JCV, HCMV, HPV and p53 codon 72 polymorphism in lung carcinoma. Lung Cancer 57:273-281.
Guo XZ, Song CL (2002) The regulation of herbal application for lung cancer. Liaoning Journal of Traditional Chinese Medicine 29:273.
Hackshaw AK, Law MR, Wald NJ (1997) The accumulated evidence on lung cancer and environmental tobacco smoke. BMJ 315:980-988.
Hamilton W, Peters TJ, Round A, Sharp D (2005) What are the clinical features of lung cancer before the diagnosis is made? A population based case-control study. Thorax 60:1059-1065.
Heald R, McKeon F (1990) Mutations of phosphorylation sites in lamin A that prevent nuclear lamina disassembly in mitosis. Cell 61:579-589.
Hecht SS (2003) Tobacco carcinogens, their biomarkers and tobacco-induced cancer. Nat Rev Cancer 3:733-744.
Herbst RS, Heymach JV, Lippman SM (2008) Lung cancer. N Engl J Med 359:1367-1380.
Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, Bruns CJ, Heeschen C (2007) Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1:313-323.
Ho MM, Ng AV, Lam S, Hung JY (2007) Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res 67:4827-4833.
Ho YT, Yang JS, Li TC, Lin JJ, Lin JG, Lai KC, Ma CY, Wood WG, Chung JG (2009a) Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-kappaB, u-PA and MMP-2 and -9. Cancer Lett 279:155-162.
Ho YT, Lu CC, Yang JS, Chiang JH, Li TC, Ip SW, Hsia TC, Liao CL, Lin JG, Wood WG, Chung JG (2009b) Berberine induced apoptosis via promoting the expression of caspase-8, -9 and -3, apoptosis-inducing factor and endonuclease G in SCC-4 human tongue squamous carcinoma cancer cells. Anticancer Res 29:4063-4070.
Hsu TS, Chen C, Lee PT, Chiu SJ, Liu HF, Tsai CC, Chao JI (2008) 7-Chloro-6-piperidin-1-yl-quinoline-5,8-dione (PT-262), a novel synthetic compound induces lung carcinoma cell death associated with inhibiting ERK and CDC2 phosphorylation via a p53-independent pathway. Cancer Chemother Pharmacol 62:799-808.
Hsu WH, Hsieh YS, Kuo HC, Teng CY, Huang HI, Wang CJ, Yang SF, Liou YS, Kuo WH (2007) Berberine induces apoptosis in SW620 human colonic carcinoma cells through generation of reactive oxygen species and activation of JNK/p38 MAPK and FasL. Arch Toxicol 81:719-728.
Hsu YL, Cho CY, Kuo PL, Huang YT, Lin CC (2006) Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) induces apoptosis and cell cycle arrest in A549 cells through p53 accumulation via c-Jun NH2-terminal kinase-mediated phosphorylation at serine 15 in vitro and in vivo. J Pharmacol Exp Ther 318:484-494.
Huang D, Gao Q, Guo L, Zhang C, Jiang W, Li H, Wang J, Han X, Shi Y, Lu SH (2009) Isolation and identification of cancer stem-like cells in esophageal carcinoma cell lines. Stem Cells Dev 18:465-473.
Huang R, Liu YG, Lin Y, Fan Y, Boynton A, Yang D, Huang RP (2001) Enhanced apoptosis under low serum conditions in human glioblastoma cells by connexin 43 (Cx43). Mol Carcinog 32:128-138.
Huber MA, Kraut N, Beug H (2005) Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 17:548-558.
Iizuka N, Miyamoto K, Okita K, Tangoku A, Hayashi H, Yosino S, Abe T, Morioka T, Hazama S, Oka M (2000) Inhibitory effect of Coptidis Rhizoma and berberine on the proliferation of human esophageal cancer cell lines. Cancer Lett 148:19-25.
Ishizawa K, Rasheed ZA, Karisch R, Wang Q, Kowalski J, Susky E, Pereira K, Karamboulas C, Moghal N, Rajeshkumar NV, Hidalgo M, Tsao M, Ailles L, Waddell TK, Maitra A, Neel BG, Matsui W (2010) Tumor-initiating cells are rare in many human tumors. Cell Stem Cell 7:279-282.
James MA, Fu H, Liu Y, Chen DR, You M (2011) Dietary administration of berberine or Phellodendron amurense extract inhibits cell cycle progression and lung tumorigenesis. Mol Carcinog 50:1-7.
Kabir Z, Bennett K, Clancy L (2007) Lung cancer and urban air-pollution in Dublin: a temporal association? Ir Med J 100:367-369.
Kang NJ, Lee KW, Kim BH, Bode AM, Lee HJ, Heo YS, Boardman L, Limburg P, Dong Z (2011) Coffee phenolic phytochemicals suppress colon cancer metastasis by targeting MEK and TOPK. Carcinogenesis.
Khleif SN, DeGregori J, Yee CL, Otterson GA, Kaye FJ, Nevins JR, Howley PM (1996) Inhibition of cyclin D-CDK4/CDK6 activity is associated with an E2F-mediated induction of cyclin kinase inhibitor activity. Proc Natl Acad Sci U S A 93:4350-4354.
Kimura K, Hirano M, Kobayashi R, Hirano T (1998) Phosphorylation and activation of 13S condensin by Cdc2 in vitro. Science 282:487-490.
Lang SH, Frame FM, Collins AT (2009) Prostate cancer stem cells. J Pathol 217:299-306.
Lee JW, Min HY, Han AR, Chung HJ, Park EJ, Park HJ, Hong JY, Seo EK, Lee SK (2007) Growth inhibition and induction of G1 phase cell cycle arrest in human lung cancer cells by a phenylbutenoid dimer isolated from Zingiber cassumunar. Biol Pharm Bull 30:1561-1564.
Li C, Heidt DG, Dalerba P, Burant CF, Zhang L, Adsay V, Wicha M, Clarke MF, Simeone DM (2007) Identification of pancreatic cancer stem cells. Cancer Res 67:1030-1037.
Li PW (2001) Comprehensive diagnosis and treatment for LC. Beijing: China Chinese Medicine Press.
Li Q, Cheng H, Zhu G, Yang L, Zhou A, Wang X, Fang N, Xia L, Su J, Wang M, Peng D, Xu Q (2010) Gambogenic acid inhibits proliferation of A549 cells through apoptosis-inducing and cell cycle arresting. Biol Pharm Bull 33:415-420.
Lin CC, Ng LT, Hsu FF, Shieh DE, Chiang LC (2004) Cytotoxic effects of Coptis chinensis and Epimedium sagittatum extracts and their major constituents (berberine, coptisine and icariin) on hepatoma and leukaemia cell growth. Clin Exp Pharmacol Physiol 31:65-69.
Lin RK, Hsieh YS, Lin P, Hsu HS, Chen CY, Tang YA, Lee CF, Wang YC (2010) The tobacco-specific carcinogen NNK induces DNA methyltransferase 1 accumulation and tumor suppressor gene hypermethylation in mice and lung cancer patients. J Clin Invest 120:521-532.
Liu CA, Peng M (1994) Dictionary of Chinese anti-cancer herbal medicine. Hubei: Hubei Science and Technology Press.
Liu JJ, Huang TS, Cheng WF, Lu FJ (2003) Baicalein and baicalin are potent inhibitors of angiogenesis: Inhibition of endothelial cell proliferation, migration and differentiation. Int J Cancer 106:559-565.
Liu S, Ma Z, Cai H, Li Q, Rong W, Kawano M (2010) Inhibitory effect of baicalein on IL-6-mediated signaling cascades in human myeloma cells. Eur J Haematol 84:137-144.
Ma Z, Otsuyama K, Liu S, Abroun S, Ishikawa H, Tsuyama N, Obata M, Li FJ, Zheng X, Maki Y, Miyamoto K, Kawano MM (2005) Baicalein, a component of Scutellaria radix from Huang-Lian-Jie-Du-Tang (HLJDT), leads to suppression of proliferation and induction of apoptosis in human myeloma cells. Blood 105:3312-3318.
Mahller YY, Williams JP, Baird WH, Mitton B, Grossheim J, Saeki Y, Cancelas JA, Ratner N, Cripe TP (2009) Neuroblastoma cell lines contain pluripotent tumor initiating cells that are susceptible to a targeted oncolytic virus. PLoS One 4:e4235.
Maitland NJ, Collins AT (2008) Prostate cancer stem cells: a new target for therapy. J Clin Oncol 26:2862-2870.
Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, Campbell LL, Polyak K, Brisken C, Yang J, Weinberg RA (2008) The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 133:704-715.
Moritsugu KP (2007) The 2006 Report of the Surgeon General: the health consequences of involuntary exposure to tobacco smoke. Am J Prev Med 32:542-543.
Mu R, Qi Q, Gu H, Wang J, Yang Y, Rong J, Liu W, Lu N, You Q, Guo Q (2009) Involvement of p53 in oroxylin A-induced apoptosis in cancer cells. Mol Carcinog 48:1159-1169.
Murray AW (2004) Recycling the cell cycle: cyclins revisited. Cell 116:221-234.
Ning ZF, Huang YJ, Lin TX, Zhou YX, Jiang C, Xu KW, Huang H, Yin XB, Huang J (2009) Subpopulations of stem-like cells in side population cells from the human bladder transitional cell cancer cell line T24. J Int Med Res 37:621-630.
O'Brien CA, Pollett A, Gallinger S, Dick JE (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445:106-110.
O'Reilly KM, McLaughlin AM, Beckett WS, Sime PJ (2007) Asbestos-related lung disease. Am Fam Physician 75:683-688.
Parada-Turska J, Paduch R, Majdan M, Kandefer-Szerszen M, Rzeski W (2007) Antiproliferative activity of parthenolide against three human cancer cell lines and human umbilical vein endothelial cells. Pharmacol Rep 59:233-237.
Park CH, Bergsagel DE, McCulloch EA (1971) Mouse myeloma tumor stem cells: a primary cell culture assay. J Natl Cancer Inst 46:411-422.
Park TY, Park MH, Shin WC, Rhee MH, Seo DW, Cho JY, Kim HM (2008) Anti-metastatic potential of ginsenoside Rp1, a novel ginsenoside derivative. Biol Pharm Bull 31:1802-1805.
Patel M, Lu L, Zander DS, Sreerama L, Coco D, Moreb JS (2008) ALDH1A1 and ALDH3A1 expression in lung cancers: correlation with histologic type and potential precursors. Lung Cancer 59:340-349.
Peacock CD, Watkins DN (2008) Cancer stem cells and the ontogeny of lung cancer. J Clin Oncol 26:2883-2889.
Peng PL, Hsieh YS, Wang CJ, Hsu JL, Chou FP (2006) Inhibitory effect of berberine on the invasion of human lung cancer cells via decreased productions of urokinase-plasminogen activator and matrix metalloproteinase-2. Toxicol Appl Pharmacol 214:8-15.
Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci U S A 104:973-978.
Seigel GM, Campbell LM, Narayan M, Gonzalez-Fernandez F (2005) Cancer stem cell characteristics in retinoblastoma. Mol Vis 11:729-737.
Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, Henkelman RM, Cusimano MD, Dirks PB (2004) Identification of human brain tumour initiating cells. Nature 432:396-401.
Sopori M (2002) Effects of cigarette smoke on the immune system. Nat Rev Immunol 2:372-377.
Sung JM, Cho HJ, Yi H, Lee CH, Kim HS, Kim DK, Abd El-Aty AM, Kim JS, Landowski CP, Hediger MA, Shin HC (2008) Characterization of a stem cell population in lung cancer A549 cells. Biochem Biophys Res Commun 371:163-167.
Teng Y, Wang X, Wang Y, Ma D (2010) Wnt/beta-catenin signaling regulates cancer stem cells in lung cancer A549 cells. Biochem Biophys Res Commun 392:373-379.
Thiery JP (2003) Epithelial-mesenchymal transitions in development and pathologies. Curr Opin Cell Biol 15:740-746.
Tirino V, Desiderio V, d'Aquino R, De Francesco F, Pirozzi G, Graziano A, Galderisi U, Cavaliere C, De Rosa A, Papaccio G, Giordano A (2008) Detection and characterization of CD133+ cancer stem cells in human solid tumours. PLoS One 3:e3469.
Travis WD (2002) Pathology of lung cancer. Clin Chest Med 23:65-81, viii.
Travis WD, Travis LB, Devesa SS (1995) Lung cancer. Cancer 75:191-202.
Wakeling AE, Guy SP, Woodburn JR, Ashton SE, Curry BJ, Barker AJ, Gibson KH (2002) ZD1839 (Iressa): an orally active inhibitor of epidermal growth factor signaling with potential for cancer therapy. Cancer Res 62:5749-5754.
Wang J, Guo LP, Chen LZ, Zeng YX, Lu SH (2007) Identification of cancer stem cell-like side population cells in human nasopharyngeal carcinoma cell line. Cancer Res 67:3716-3724.
WHO (2009) Cancer. In: World Health Organization.
Zhang HZ, Lin XG, Hua P, Wang M, Ao X, Xiong LH, Wu C, Guo JJ (2010) The study of the tumor stem cell properties of CD133+CD44+ cells in the human lung adenocarcinoma cell line A549. Cell Mol Biol (Noisy-le-grand) 56 Suppl:OL1350-1358.
Zhang S, Balch C, Chan MW, Lai HC, Matei D, Schilder JM, Yan PS, Huang TH, Nephew KP (2008) Identification and characterization of ovarian cancer-initiating cells from primary human tumors. Cancer Res 68:4311-4320.
Zhang WD, Zhao HR, Yan Y, Wang XH, Zong ZH, Liu Y (2005) [Apoptosis induced by cantharidin in human pulmonary carcinoma cells A549 and its molecular mechanisms]. Zhonghua Zhong Liu Za Zhi 27:330-334.
Zheng H, Abdel Aziz HO, Nakanishi Y, Masuda S, Saito H, Tsuneyama K, Takano Y (2007) Oncogenic role of JC virus in lung cancer. J Pathol 212:306-315.