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研究生:鄭仲惠
研究生(外文):Jhong-Huei Jheng
論文名稱:在人類肺癌細胞中甘胺酸螯合鐵透過抑制低氧誘導因子來調控細胞的能量代謝及侵襲性
論文名稱(外文):Ferrous glycinate regulates cell energy metabolism and invasiveness via suppression of hypoxia-induced factor in human A549 cells
指導教授:李宏謨李宏謨引用關係
指導教授(外文):Horng-Mo Lee
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:醫學檢驗暨生物技術學系所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2016
畢業學年度:104
語文別:中文
論文頁數:82
中文關鍵詞:低氧誘導因子瓦氏效應甘胺酸螯合鐵能量代謝細胞侵襲性
外文關鍵詞:HIF-1αWarburg effecferrous glycinateenergy metabolismcell invasiveness
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低氧誘導因子(Hypoxia-inducible factor 1 alpha, HIF-1α) 在實質性腫瘤的缺氧微環境中扮演重要角色,調控了癌細胞的增殖,遷移性和侵襲性。而在目前的研究結果中,我們發現甘胺酸螯合鐵會透過活化脯氨酰羥化酶(prolyl hydroxylase)降低HIF-1α的蛋白表現量,進而調控葡萄糖轉運蛋白(Glucose transporter-1, Glut-1)、己糖激酶(hexokinase-2)、乳酸脫氫酶(lactate dehydrogenase A, LDHA)而降低有氧醣解作用。另一方面,也減少了丙酮酸脫氫酶激酶(pyruvate dehydrogenase kinase, PDK)的蛋白表現量及丙酮酸脫氫酶(pyruvate dehydrogenase, PDH)的磷酸化,進而增加粒線體膜電位和ATP的產生,因此推測甘胺酸螯合鐵可能反轉了癌細胞的瓦氏效應(Warburg effect)。此外也發現甘胺酸螯合鐵能夠減少Twist蛋白量,導致增加E-cadherin蛋白量及減少Vimentin蛋白量,改善了癌細胞的上皮細胞間質轉化現象,進而抑制細胞的遷移速度及侵襲性。這些結果表示甘胺酸螯合鐵可能可以成為治療人類肺腺癌的佐劑。

HIF-1α is a multi-facet protein that control tumor progression, including cell proliferation, invasion and metastasis. In the present study, we demonstrated that incubation of A549 cells with ferrous glycinate decreased the protein levels of HIF-1α through a prolyl hydroxylation-dependent mechanism. The reduction of HIF-1α level was associated with decreased expression levels of glucose transporter, Glut-1, and glycolytic enzymes including hexokinase-2, and lactate dehydrogenase A. On the other hand, treatment with ferrous glycinate reactivated oxidative phosphorylation by decreasing the expression of pyruvate dehydrogenase kinase-1 and pyruvate dehydrogenase phosphorylation that subsequently increased mitochondrial membrane potential and ATP production, suggesting ferrous glycinate may reverse Warburg effect. In addition, ferrous glycinate decreased cell migration and invasion in A549 cells, which was accompanied with reduced twist and vimentin protein levels and increased of E-cadherin, suggesting ferrous glycinate may alter the epithelial mesenchymal transition. These results indicate that ferrous glycinate may serve as a therapeutic adjuvant for human lung adenocarcinoma.

目錄 I
圖目錄 IV
縮寫表 VI
Abstract 1
中文摘要 2
文獻回顧(Introduction) 3
肺癌(Lung cancer) 4
瓦氏效應(Warburg effect):腫瘤細胞的醣類代謝 4
上皮細胞間質轉化(Epithelial-mesenchymal transition, EMT) 5
低氧誘導因子(Hypoxia-inducible factor 1-alpha, HIF-1α) 7
研究目的 9
實驗材料與方法(Material and Methods) 12
材料(Material) 13
人類肺腺癌上皮細胞培養(A549 cell culture) 18
細胞增生(Cell proliferation) 18
細胞蛋白質測定(Protein assay) 18
西方墨點法(Western blotting) 19
乳酸測定(Lactate assay) 20
ATP測定(ATP assay) 21
粒線體膜電位測定(JC-1 assay) 21
細胞遷移(Migration) 22
明膠蛋白酶活性分析(Gelatin zymography) 22
細胞侵襲(Invasion assay) 22
軟性瓊膠群落生長試驗(Soft agar growth assay) 23
統計方式 24
實驗結果(Result) 25
甘胺酸螯合鐵在人類肺腺癌上皮細胞中降低了HIF-1α蛋白量 26
甘胺酸螯合鐵降低人類肺腺癌上皮細胞的醣解作用 27
甘胺酸螯合鐵恢復人類肺腺癌上皮細胞的粒線體功能 28
甘胺酸螯合鐵改善人類肺腺癌上皮細胞的上皮細胞間質轉化現象 30
討論(Discussion) 32
參考文獻 37
附圖 43


Agani F, Jiang BH (2013) Oxygen-independent regulation of HIF-1: novel involvement of PI3K/AKT/mTOR pathway in cancer. Curr Cancer Drug Targets 13: 245-51

Bosutti A, Qi J, Pennucci R, Bolton D, Matou S, Ali K, Tsai LH, Krupinski J, Petcu EB, Montaner J, Al Baradie R, Caccuri F, Caruso A, Alessandri G, Kumar S, Rodriguez C, Martinez-Gonzalez J, Slevin M (2013) Targeting p35/Cdk5 signalling via CIP-peptide promotes angiogenesis in hypoxia. PLoS One 8: e75538

Burroughs SK, Kaluz S, Wang D, Wang K, Van Meir EG, Wang B (2013) Hypoxia inducible factor pathway inhibitors as anticancer therapeutics. Future Med Chem 5: 553-72

Cannito S, Novo E, Compagnone A, Valfre di Bonzo L, Busletta C, Zamara E, Paternostro C, Povero D, Bandino A, Bozzo F, Cravanzola C, Bravoco V, Colombatto S, Parola M (2008) Redox mechanisms switch on hypoxia-dependent epithelial-mesenchymal transition in cancer cells. Carcinogenesis 29: 2267-78

Chen S, Sang N (2016) Hypoxia-Inducible Factor-1: A Critical Player in the Survival Strategy of Stressed Cells. J Cell Biochem 117: 267-78

Cohen R, Neuzillet C, Tijeras-Raballand A, Faivre S, de Gramont A, Raymond E (2015) Targeting cancer cell metabolism in pancreatic adenocarcinoma. Oncotarget 6: 16832-47

Denko NC (2008) Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer 8: 705-13

Du J, Sun B, Zhao X, Gu Q, Dong X, Mo J, Sun T, Wang J, Sun R, Liu Y (2014) Hypoxia promotes vasculogenic mimicry formation by inducing epithelial-mesenchymal transition in ovarian carcinoma. Gynecol Oncol 133: 575-83

Gambhir SS (2002) Molecular imaging of cancer with positron emission tomography. Nat Rev Cancer 2: 683-93

Hayashi M, Fukuhara H, Inoue K, Shuin T, Hagiya Y, Nakajima M, Tanaka T, Ogura S (2015) The effect of iron ion on the specificity of photodynamic therapy with 5-aminolevulinic acid. PLoS One 10: e0122351

Hirsila M, Koivunen P, Gunzler V, Kivirikko KI, Myllyharju J (2003) Characterization of the human prolyl 4-hydroxylases that modify the hypoxia-inducible factor. J Biol Chem 278: 30772-80

Hockel M, Vaupel P (2001) Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects. J Natl Cancer Inst 93: 266-76

Hotary K, Li XY, Allen E, Stevens SL, Weiss SJ (2006) A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev 20: 2673-86

Ioannou M, Paraskeva E, Baxevanidou K, Simos G, Papamichali R, Papacharalambous C, Samara M, Koukoulis G (2015) HIF-1alpha in colorectal carcinoma: review of the literature. J BUON 20: 680-9

Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M, Salic A, Asara JM, Lane WS, Kaelin WG, Jr. (2001) HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 292: 464-8

Iyer NV, Kotch LE, Agani F, Leung SW, Laughner E, Wenger RH, Gassmann M, Gearhart JD, Lawler AM, Yu AY, Semenza GL (1998) Cellular and developmental control of O2 homeostasis by hypoxia-inducible factor 1 alpha. Genes Dev 12: 149-62

Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ, von Kriegsheim A, Hebestreit HF, Mukherji M, Schofield CJ, Maxwell PH, Pugh CW, Ratcliffe PJ (2001) Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science 292: 468-72

Kalluri R, Neilson EG (2003) Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest 112: 1776-84

Katayama A, Bandoh N, Kishibe K, Takahara M, Ogino T, Nonaka S, Harabuchi Y (2004) Expressions of matrix metalloproteinases in early-stage oral squamous cell carcinoma as predictive indicators for tumor metastases and prognosis. Clin Cancer Res 10: 634-40

Kim JW, Tchernyshyov I, Semenza GL, Dang CV (2006) HIF-1-mediated expression of pyruvate dehydrogenase kinase: a metabolic switch required for cellular adaptation to hypoxia. Cell Metab 3: 177-85

Lai JH, Jan HJ, Liu LW, Lee CC, Wang SG, Hueng DY, Cheng YY, Lee HM, Ma HI (2013) Nodal regulates energy metabolism in glioma cells by inducing expression of hypoxia-inducible factor 1alpha. Neuro Oncol 15: 1330-41

Laughner E, Taghavi P, Chiles K, Mahon PC, Semenza GL (2001) HER2 (neu) signaling increases the rate of hypoxia-inducible factor 1alpha (HIF-1alpha) synthesis: novel mechanism for HIF-1-mediated vascular endothelial growth factor expression. Mol Cell Biol 21: 3995-4004

Liu T, Zhang X, Shang M, Zhang Y, Xia B, Niu M, Liu Y, Pang D (2013) Dysregulated expression of Slug, vimentin, and E-cadherin correlates with poor clinical outcome in patients with basal-like breast cancer. J Surg Oncol 107: 188-94

Liu Y, Cui Y, Shi M, Zhang Q, Wang Q, Chen X (2014) Deferoxamine promotes MDA-MB-231 cell migration and invasion through increased ROS-dependent HIF-1alpha accumulation. Cell Physiol Biochem 33: 1036-46

Lu H, Forbes RA, Verma A (2002) Hypoxia-inducible factor 1 activation by aerobic glycolysis implicates the Warburg effect in carcinogenesis. J Biol Chem 277: 23111-5

Maccio A, Madeddu C, Gramignano G, Mulas C, Tanca L, Cherchi MC, Floris C, Omoto I, Barracca A, Ganz T (2015) The role of inflammation, iron, and nutritional status in cancer-related anemia: results of a large, prospective, observational study. Haematologica 100: 124-32

Manz DH, Blanchette NL, Paul BT, Torti FM, Torti SV (2016) Iron and cancer: recent insights. Ann N Y Acad Sci 1368: 149-61

Maxwell PH, Wiesener MS, Chang GW, Clifford SC, Vaux EC, Cockman ME, Wykoff CC, Pugh CW, Maher ER, Ratcliffe PJ (1999) The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature 399: 271-5

Mole DR (2010) Iron homeostasis and its interaction with prolyl hydroxylases. Antioxid Redox Signal 12: 445-58

Ohh M, Park CW, Ivan M, Hoffman MA, Kim TY, Huang LE, Pavletich N, Chau V, Kaelin WG (2000) Ubiquitination of hypoxia-inducible factor requires direct binding to the beta-domain of the von Hippel-Lindau protein. Nat Cell Biol 2: 423-7

Onder TT, Gupta PB, Mani SA, Yang J, Lander ES, Weinberg RA (2008) Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res 68: 3645-54

Onnis B, Rapisarda A, Melillo G (2009) Development of HIF-1 inhibitors for cancer therapy. J Cell Mol Med 13: 2780-6

Paoli P, Giannoni E, Chiarugi P (2013) Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta 1833: 3481-98

Rankin EB, Giaccia AJ (2008) The role of hypoxia-inducible factors in tumorigenesis. Cell Death Differ 15: 678-85

Rennebeck G, Martelli M, Kyprianou N (2005) Anoikis and survival connections in the tumor microenvironment: is there a role in prostate cancer metastasis? Cancer Res 65: 11230-5

Rensvold JW, Ong SE, Jeevananthan A, Carr SA, Mootha VK, Pagliarini DJ (2013) Complementary RNA and protein profiling identifies iron as a key regulator of mitochondrial biogenesis. Cell Rep 3: 237-45

Semenza GL (2003) Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3: 721-32

Semenza GL, Roth PH, Fang HM, Wang GL (1994) Transcriptional regulation of genes encoding glycolytic enzymes by hypoxia-inducible factor 1. J Biol Chem 269: 23757-63
Siegert I, Schodel J, Nairz M, Schatz V, Dettmer K, Dick C, Kalucka J, Franke K, Ehrenschwender M, Schley G, Beneke A, Sutter J, Moll M, Hellerbrand C, Wielockx B, Katschinski DM, Lang R, Galy B, Hentze MW, Koivunen P et al. (2015) Ferritin-Mediated Iron Sequestration Stabilizes Hypoxia-Inducible Factor-1alpha upon LPS Activation in the Presence of Ample Oxygen. Cell Rep 13: 2048-55

Tong WH, Sourbier C, Kovtunovych G, Jeong SY, Vira M, Ghosh M, Romero VV, Sougrat R, Vaulont S, Viollet B, Kim YS, Lee S, Trepel J, Srinivasan R, Bratslavsky G, Yang Y, Linehan WM, Rouault TA (2011) The glycolytic shift in fumarate-hydratase-deficient kidney cancer lowers AMPK levels, increases anabolic propensities and lowers cellular iron levels. Cancer Cell 20: 315-27

Torti SV, Torti FM (2013) Iron and cancer: more ore to be mined. Nat Rev Cancer 13: 342-55

Tsai JH, Yang J (2013) Epithelial-mesenchymal plasticity in carcinoma metastasis. Genes Dev 27: 2192-206

Warburg O (1956) On the origin of cancer cells. Science 123: 309-14

Wu M, Neilson A, Swift AL, Moran R, Tamagnine J, Parslow D, Armistead S, Lemire K, Orrell J, Teich J, Chomicz S, Ferrick DA (2007) Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. Am J Physiol Cell Physiol 292: C125-36

Xiao D, He J (2010) Epithelial mesenchymal transition and lung cancer. J Thorac Dis 2: 154-9

Yang MH, Wu MZ, Chiou SH, Chen PM, Chang SY, Liu CJ, Teng SC, Wu KJ (2008) Direct regulation of TWIST by HIF-1alpha promotes metastasis. Nat Cell Biol 10: 295-305

Zavadil J, Haley J, Kalluri R, Muthuswamy SK, Thompson E (2008) Epithelial-mesenchymal transition. Cancer Res 68: 9574-7

Zhang W, Wu Y, Yan Q, Ma F, Shi X, Zhao Y, Peng Y, Wang J, Jiang B (2014) Deferoxamine enhances cell migration and invasion through promotion of HIF-1alpha expression and epithelial-mesenchymal transition in colorectal cancer. Oncol Rep 31: 111-6


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