研究生: |
韋會妤 Hui-Yu Wei |
---|---|
論文名稱: |
利用非小細胞肺癌細胞以及動物模式探討中草藥之治療潛力 Investigation of potential Chinese herbs in non-small cell lung carcinoma in vitro and in vivo |
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: | 2013 |
畢業學年度: | 101 |
語文別: | 中文 |
論文頁數: | 74 |
中文關鍵詞: | 非小細胞肺癌 、中藥 、異種移植 、細胞週期 、細胞凋亡 |
英文關鍵詞: | NSCLC, CHM, xenograft model, cell cycle, apoptosis |
論文種類: | 學術論文 |
相關次數: | 點閱:155 下載:4 |
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非小細胞肺癌在肺癌中具有高發生率與致死率。文獻報導有許多中草藥含抑制腫瘤生長、血管新生和轉移的能力,同時伴隨較少的副作用。我們先前從十五種臨床上常用的中藥複方中,篩檢出三種對非小細胞肺癌細胞具有抑制能力的複方,分別為三號、五號及十四號藥方。已知此三種複方會造成A549細胞週期停滯進而抑制細胞的生長。我們更進一步使用非小細胞肺癌細胞株H460以及H520為細胞模式,評估這三種複方是否對非小細胞肺癌細胞具有一致的抑制效果,並以A549細胞株於雄性裸鼠進行腫瘤異種移植動物模式,以評估藥方的抗癌效果。在細胞模式中,我們利用細胞存活率(MTT assay)檢測十四號複方對非小細胞肺癌細胞株H460以及H520的半致死率,分別為4.5% 跟3.8%。此外,我們也發現十四號複方具有抑制細胞群落形成,以及造成細胞週期停滯在G1與G2的現象。在異種移植動物模式中,我們投予三種複方後發現三號以及十四號複方可減少腫瘤的大小,以西方墨點轉漬法進一步探討複方藥物處理的結果,我們發現這兩種複方可能藉由將腫瘤內細胞停滯在G2細胞週期以及誘導細胞凋亡的發生。本實驗的結果提供細胞以及動物實驗模式的結果顯示三號以及十四號複方可能具有抗腫瘤的潛力。
Non-small cell lung carcinoma (NSCLC) is lung cancer with the highest incidence and mortality rate in the world. Some Chinese herb medicines (CHM) have been reported to have potential anti-cancer activity through reducing tumor recurrence, angiogenesis, metastasis, and drug-resistance.
Our previous studies have identified Formulae 3, 5, and14, showing potential anti-cancer activity in NSCLC cell lines. These formulae were found to have anti-proliferative activity through G1 and G2 arrest toward the A549 cells. We further investigated the effect of these formulae toward another two NSCLC cancer cell lines. Using MTT assay, we have determined the IC50 of Formula 14 in NSCLC H460 and H520 to be 4.5% and 3.8%, respectively. Furthermore, Formula 14 reduced the ability of colony formation, and induced G1 and G2 arrest.
In addition, we characterized their therapeutic potential in vivo using mouse xenograft tumor model. We confirmed that all the 3 formula treatment had no notable toxicity for the mice though the pathological examination of mouse kidney and liver. We found that tumor size was decreased by Formulae 3 and 14. Further investigation showed that these formulae may induce G2 arrest and apoptosis to reduce the tumor size. In conclusion, we found that Formulae 3 and 14 showed potential anti-cancer activity from both the in vitro and in vivo analyses.
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. The Journal of clinical investigation 119:1438-1449.
Agarwal ML, Agarwal A, Taylor WR, Chernova O, Sharma Y, Stark GR (1998) A p53-dependent S-phase checkpoint helps to protect cells from DNA damage in response to starvation for pyrimidine nucleotides. Proc Natl Acad Sci U S A 95:14775-14780.
Albers JW, Chaudhry V, Cavaletti G, Donehower RC (2011) Interventions for preventing neuropathy caused by cisplatin and related compounds. Cochrane Database Syst Rev CD005228.
Attardi LD, Lowe SW, Brugarolas J, Jacks T (1996) Transcriptional activation by p53, but not induction of the p21 gene, is essential for oncogene-mediated apoptosis. EMBO J 15:3693-3701.
Badie C, Bourhis J, Sobczak-Thepot J, Haddada H, Chiron M, Janicot M, Janot F, Tursz T, Vassal G (2000) p53-dependent G2 arrest associated with a decrease in cyclins A2 and B1 levels in a human carcinoma cell line. Br J Cancer 82:642-650.
Banks-Schlegel SP, Gazdar AF, Harris CC (1985) Intermediate filament and cross-linked envelope expression in human lung tumor cell lines. Cancer research 45:1187-1197.
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.
Chao JC, Chiang SW, Wang CC, Tsai YH, Wu MS (2006) Hot water-extracted Lycium barbarum and Rehmannia glutinosa inhibit proliferation and induce apoptosis of hepatocellular carcinoma cells. World J Gastroenterol 12:4478-4484.
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.
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.
Choi HJ, Seon MR, Lim SS, Kim JS, Chun HS, Park JH (2008) Hexane/ethanol extract of Glycyrrhiza uralensis licorice suppresses doxorubicin-induced apoptosis in H9c2 rat cardiac myoblasts. Exp Biol Med (Maywood) 233:1554-1560.
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.
Dulic V, Kaufmann WK, Wilson SJ, Tlsty TD, Lees E, Harper JW, Elledge SJ, Reed SI (1994) p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest. Cell 76:1013-1023.
Evan GI, Brown L, Whyte M, Harrington E (1995) Apoptosis and the cell cycle. Current opinion in cell biology 7:825-834.
Gao J, Morgan WA, Sanchez-Medina A, Corcoran O (2011) The ethanol extract of Scutellaria baicalensis and the active compounds induce cell cycle arrest and apoptosis including upregulation of p53 and Bax in human lung cancer cells. Toxicology and applied pharmacology 254:221-228.
Gaube F, Wolfl S, Pusch L, Kroll TC, Hamburger M (2007) Gene expression profiling reveals effects of Cimicifuga racemosa (L.) NUTT. (black cohosh) on the estrogen receptor positive human breast cancer cell line MCF-7. BMC Pharmacol 7:11.
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.
Gu H, Wang X, Rao S, Wang J, Zhao J, Ren FL, Mu R, Yang Y, Qi Q, Liu W, Lu N, Ling H, You Q, Guo Q (2008) Gambogic acid mediates apoptosis as a p53 inducer through down-regulation of mdm2 in wild-type p53-expressing cancer cells. Mol Cancer Ther 7:3298-3305.
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.
Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ (1993) The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75:805-816.
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.
Hostanska K, Nisslein T, Freudenstein J, Reichling J, Saller R (2004) Cimicifuga racemosa extract inhibits proliferation of estrogen receptor-positive and negative human breast carcinoma cell lines by induction of apoptosis. Breast Cancer Res Treat 84:151-160.
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.
Hsu YL, Kuo PL, Tzeng TF, Sung SC, Yen MH, Lin LT, Lin CC (2008) Huang-lian-jie-du-tang, a traditional Chinese medicine prescription, induces cell-cycle arrest and apoptosis in human liver cancer cells in vitro and in vivo. J Gastroenterol Hepatol 23:e290-299.
Huber MA, Kraut N, Beug H (2005) Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 17:548-558.
Ireland H, Kemp R, Houghton C, Howard L, Clarke AR, Sansom OJ, Winton DJ (2004) Inducible Cre-mediated control of gene expression in the murine gastrointestinal tract: effect of loss of beta-catenin. Gastroenterology 126:1236-1246.
Jiang F, Qiu Q, Khanna A, Todd NW, Deepak J, Xing L, Wang H, Liu Z, Su Y, Stass SA, Katz RL (2009) Aldehyde dehydrogenase 1 is a tumor stem cell-associated marker in lung cancer. Molecular cancer research : MCR 7:330-338.
Kabir Z, Bennett K, Clancy L (2007) Lung cancer and urban air-pollution in Dublin: a temporal association? Ir Med J 100:367-369.
Kerbel RS (2003) Human tumor xenografts as predictive preclinical models for anticancer drug activity in humans: better than commonly perceived-but they can be improved. Cancer Biol Ther 2:S134-139.
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.
Kihara A, Kabeya Y, Ohsumi Y, Yoshimori T (2001) Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network. EMBO Rep 2:330-335.
Kimura K, Hirano M, Kobayashi R, Hirano T (1998) Phosphorylation and activation of 13S condensin by Cdc2 in vitro. Science 282:487-490.
King RW, Jackson PK, Kirschner MW (1994) Mitosis in transition. Cell 79:563-571.
Koops HS, Vaglini M, Suciu S, Kroon BB, Thompson JF, Gohl J, Eggermont AM, Di Filippo F, Krementz ET, Ruiter D, Lejeune FJ (1998) Prophylactic isolated limb perfusion for localized, high-risk limb melanoma: results of a multicenter randomized phase III trial. European Organization for Research and Treatment of Cancer Malignant Melanoma Cooperative Group Protocol 18832, the World Health Organization Melanoma Program Trial 15, and the North American Perfusion Group Southwest Oncology Group-8593. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 16:2906-2912.
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.
Leonard CJ, Canman CE, Kastan MB (1995) The role of p53 in cell-cycle control and apoptosis: implications for cancer. Important advances in oncology 33-42.
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.
Li R, Waga S, Hannon GJ, Beach D, Stillman B (1994) Differential effects by the p21 CDK inhibitor on PCNA-dependent DNA replication and repair. Nature 371:534-537.
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. Tumor Index 31-33.
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.
McGowan CH, Russell P (1993) Human Wee1 kinase inhibits cell division by phosphorylating p34cdc2 exclusively on Tyr15. EMBO J 12:75-85.
Mitchell KO, Ricci MS, Miyashita T, Dicker DT, Jin Z, Reed JC, El-Deiry WS (2000) Bax is a transcriptional target and mediator of c-myc-induced apoptosis. Cancer research 60:6318-6325.
Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin HK, Liebermann DA, Hoffman B, Reed JC (1994) Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene 9:1799-1805.
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.
Morton CL, Houghton PJ (2007) Establishment of human tumor xenografts in immunodeficient mice. Nat Protoc 2:247-250.
Murray AW (2004) Recycling the cell cycle: cyclins revisited. Cell 116:221-234.
Murray CJ, Lopez AD (1997) Alternative projections of mortality and disability by cause 1990-2020: Global Burden of Disease Study. Lancet 349:1498-1504.
O'Leary DJ, Madgewick R, Wallace J, Ang J (1998) Size and number of epithelial cells washed from the cornea after contact lens wear. Optom Vis Sci 75:692-696.
O'Reilly KM, McLaughlin AM, Beckett WS, Sime PJ (2007) Asbestos-related lung disease. Am Fam Physician 75:683-688.
Oltvai ZN, Milliman CL, Korsmeyer SJ (1993) Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609-619.
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 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.
Parker LL, Piwnica-Worms H (1992) Inactivation of the p34cdc2-cyclin B complex by the human WEE1 tyrosine kinase. Science 257:1955-1957.
Peacock CD, Watkins DN (2008) Cancer stem cells and the ontogeny of lung cancer. J Clin Oncol 26:2883-2889.
Pearson AS, Spitz FR, Swisher SG, Kataoka M, Sarkiss MG, Meyn RE, McDonnell TJ, Cristiano RJ, Roth JA (2000) Up-regulation of the proapoptotic mediators Bax and Bak after adenovirus-mediated p53 gene transfer in lung cancer cells. Clin Cancer Res 6:887-890.
Qi Q, Gu H, Yang Y, Lu N, Zhao J, Liu W, Ling H, You QD, Wang X, Guo Q (2008) Involvement of matrix metalloproteinase 2 and 9 in gambogic acid induced suppression of MDA-MB-435 human breast carcinoma cell lung metastasis. J Mol Med (Berl) 86:1367-1377.
Qiang L, Yang Y, You QD, Ma YJ, Yang L, Nie FF, Gu HY, Zhao L, Lu N, Qi Q, Liu W, Wang XT, Guo QL (2008) Inhibition of glioblastoma growth and angiogenesis by gambogic acid: an in vitro and in vivo study. Biochem Pharmacol 75:1083-1092.
Ryan JJ, Prochownik E, Gottlieb CA, Apel IJ, Merino R, Nunez G, Clarke MF (1994) c-myc and bcl-2 modulate p53 function by altering p53 subcellular trafficking during the cell cycle. Proc Natl Acad Sci U S A 91:5878-5882.
Sabbatini P, Lin J, Levine AJ, White E (1995) Essential role for p53-mediated transcription in E1A-induced apoptosis. Genes Dev 9:2184-2192.
Schmelzle T, Hall MN (2000) TOR, a central controller of cell growth. Cell 103:253-262.
Schreiber M, Kolbus A, Piu F, Szabowski A, Mohle-Steinlein U, Tian J, Karin M, Angel P, Wagner EF (1999) Control of cell cycle progression by c-Jun is p53 dependent. Genes Dev 13:607-619.
Shackelford C, Long G, Wolf J, Okerberg C, Herbert R (2002) Qualitative and quantitative analysis of nonneoplastic lesions in toxicology studies. Toxicologic pathology 30:93-96.
Sherr CJ (2004) Principles of tumor suppression. Cell 116:235-246.
Smits VA, Klompmaker R, Vallenius T, Rijksen G, Makela TP, Medema RH (2000) p21 inhibits Thr161 phosphorylation of Cdc2 to enforce the G2 DNA damage checkpoint. J Biol Chem 275:30638-30643.
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.
Swisher SG, Roth JA, Nemunaitis J, Lawrence DD, Kemp BL, Carrasco CH, Connors DG, El-Naggar AK, Fossella F, Glisson BS, Hong WK, Khuri FR, Kurie JM, Lee JJ, Lee JS, Mack M, Merritt JA, Nguyen DM, Nesbitt JC, Perez-Soler R, Pisters KM, Putnam JB, Jr., Richli WR, Savin M, Schrump DS, Shin DM, Shulkin A, Walsh GL, Wait J, Weill D, Waugh MK (1999) Adenovirus-mediated p53 gene transfer in advanced non-small-cell lung cancer. Journal of the National Cancer Institute 91:763-771.
Szabo E, Riffe ME, Steinberg SM, Birrer MJ, Linnoila RI (1996) Altered cJUN expression: an early event in human lung carcinogenesis. Cancer research 56:305-315.
Tammemagi MC, McLaughlin JR, Bull SB (1999) Meta-analyses of p53 tumor suppressor gene alterations and clinicopathological features in resected lung cancers. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology 8:625-634.
Taylor EH, Fink LM, Pappas AA (1989) Reproducibility of creatine kinase isoenzyme electrophoresis. Clin Chem 35:710.
Thiery JP (2003) Epithelial-mesenchymal transitions in development and pathologies. Current opinion in cell biology 15:740-746.
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 TC, Fang CN, Shen CC, Wei HY, Weng YP, Lin JY, Hsieh-Li HM, Lee CY (2012) Yang-Dan-Tang, Identified from 15 Chinese Herbal Formulae, Inhibits Human Lung Cancer Cell Proliferation via Cell Cycle Arrest. Evidence-based complementary and alternative medicine : eCAM 2012:276032.
Wu X, Cai ZD, Lou LM, Zhu YB (2012) Expressions of p53, c-MYC, BCL-2 and apoptotic index in human osteosarcoma and their correlations with prognosis of patients. Cancer epidemiology 36:212-216.
Xiong Y, Zhang H, Beach D (1993) Subunit rearrangement of the cyclin-dependent kinases is associated with cellular transformation. Genes Dev 7:1572-1583.
Xu C, Xie D, Yu SC, Yang XJ, He LR, Yang J, Ping YF, Wang B, Yang L, Xu SL, Cui W, Wang QL, Fu WJ, Liu Q, Qian C, Cui YH, Rich JN, Kung HF, Zhang X, Bian XW (2013) beta-Catenin/POU5F1/SOX2 transcription factor complex mediates IGF-I receptor signaling and predicts poor prognosis in lung adenocarcinoma. Cancer research 73:3181-3189.
Yun J, Chae HD, Choy HE, Chung J, Yoo HS, Han MH, Shin DY (1999) p53 negatively regulates cdc2 transcription via the CCAAT-binding NF-Y transcription factor. J Biol Chem 274:29677-29682.
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.