黑色素瘤在皮膚癌中是最為惡性的細胞之一，而化療藥物順鉑是目前化療的一線用藥，然而，黑色素癌在化療藥物順鉑的治療下卻有著抗藥性，造成順鉑無法根治黑色素癌的狀況。在我們過去的研究中指出，黑色素癌細胞再經由順鉑治療後會增加其stemness marker的表現，並且增加細胞膜上幫浦ABCB5的表現量，而這抗藥性的能力可能來自於細胞內能量ATP的產生：由於順鉑造成的粒線體增多而產生的大量ATP。葡萄糖類似物2-deoxyglucose (2-DG)在過去的研究中扮演著抑制glycolysis的角色，能抑制細胞內ATP的產生。在此研究中，2-DG能降低因為順鉑增加的黑色素癌細胞中ATP的生成量，並且降低ABCB5的表現量。而在2-DG和順鉑共同治療的狀況下，黑色素癌細胞的生長能力、轉移能力皆受到更有效的抑制。更進一步，我們降低粒線體生產ATP的作用路徑，PGC-1α，希望能找出黑色素細胞瘤對順鉑的抗藥原因，以便將來應用在未來的醫療上。

Although cisplatin is among the most potent antitumor agents, cisplatin-based chemotherapy has shown limited effectiveness for treating melanoma. Our previous work showed that cisplatin induces chemoresistance in melanoma by elevating cancer stemness marker ABCB5, which indicates the involvement of chemoresistance mechanisms in the ATP synthesis pathway. Specifically, cisplatin treatment increases production of mitochondria and ATP in melanoma. The glucose analog 2-deoxyglucose (2-DG), which is a glycolytic inhibitor, decreases cellular ATP. Specifically, 2-DG attenuates the elevation of cellular ATP caused by cisplatin in B16-F10 melanoma cells. After cisplatin treatment, 2-DG also reduces cellular levels of ABCB5. Moreover, 2-DG enhances the inhibitory effect of cisplatin on anchorage-independent growth of B16-F10 melanoma cells. Invasion assays further show that a combined treatment with 2-DG (2 mM) and cisplatin (3 μM) has a larger attenuating effect on the invasiveness of B16-F10 melanoma cells compared to 2-DG alone or cisplatin alone. In conclusion, treatment combining the glycolytic inhibitor 2-DG with cisplatin may have potent applications in melanoma therapy.

Abstract in Chinese i

Abstract in English ii

Introduction 1

Materials and Methods 8

Result 13

Discussion 20

References 24

Figures and legends 29

Appendix 43

1. Jerant AF, Johnson JT, Sheridan CD, Caffrey TJ. Early detection and treatment of skin cancer. Am Fam Physician. 2000 Jul 15;62(2):357-68, 75-6, 81-2.
2. Aberdam E, Romero C, Ortonne JP. Repeated UVB irradiations do not have the same potential to promote stimulation of melanogenesis in cultured normal human melanocytes. J Cell Sci. 1993 Dec;106 ( Pt 4):1015-22.
3. Chin L, Garraway LA, Fisher DE. Malignant melanoma: genetics and therapeutics in the genomic era. Genes Dev. 2006 Aug 15;20(16):2149-82.
4. Klein CA. Cancer. The metastasis cascade. Science. 2008 Sep 26;321(5897):1785-7.
5. O'Hagan KA, Cocchiglia S, Zhdanov AV, Tambuwala MM, Cummins EP, Monfared M, et al. PGC-1alpha is coupled to HIF-1alpha-dependent gene expression by increasing mitochondrial oxygen consumption in skeletal muscle cells. Proc Natl Acad Sci U S A. 2009 Feb 17;106(7):2188-93.
6. Klimcakova E, Chenard V, McGuirk S, Germain D, Avizonis D, Muller WJ, et al. PGC-1alpha promotes the growth of ErbB2/Neu-induced mammary tumors by regulating nutrient supply. Cancer Res. 2012 Mar 15;72(6):1538-46.
7. Puigserver P, Wu Z, Park CW, Graves R, Wright M, Spiegelman BM. A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis. Cell. 1998 Mar 20;92(6):829-39.
8. Arany Z, He H, Lin J, Hoyer K, Handschin C, Toka O, et al. Transcriptional coactivator PGC-1 alpha controls the energy state and contractile function of cardiac muscle. Cell Metab. 2005 Apr;1(4):259-71.
9. Lin J, Handschin C, Spiegelman BM. Metabolic control through the PGC-1 family of transcription coactivators. Cell Metab. 2005 Jun;1(6):361-70.
10. Liang H, Ward WF. PGC-1alpha: a key regulator of energy metabolism. Adv Physiol Educ. 2006 Dec;30(4):145-51.
11. Tcherepanova I, Puigserver P, Norris JD, Spiegelman BM, McDonnell DP. Modulation of estrogen receptor-alpha transcriptional activity by the coactivator PGC-1. J Biol Chem. 2000 May 26;275(21):16302-8.
12. Pelicano H, Martin DS, Xu RH, Huang P. Glycolysis inhibition for anticancer treatment. Oncogene. 2006 Aug 7;25(34):4633-46.
13. Prestayko AW, D'Aoust JC, Issell BF, Crooke ST. Cisplatin (cis-diamminedichloroplatinum II). Cancer Treat Rev. 1979 Mar;6(1):17-39.
14. Rosenberg B, Van Camp L, Grimley EB, Thomson AJ. The inhibition of growth or cell division in Escherichia coli by different ionic species of platinum(IV) complexes. J Biol Chem. 1967 Mar 25;242(6):1347-52.
15. Pruefer FG, Lizarraga F, Maldonado V, Melendez-Zajgla J. Participation of Omi Htra2 serine-protease activity in the apoptosis induced by cisplatin on SW480 colon cancer cells. J Chemother. 2008 Jun;20(3):348-54.
16. Chen KG, Valencia JC, Gillet JP, Hearing VJ, Gottesman MM. Involvement of ABC transporters in melanogenesis and the development of multidrug resistance of melanoma. Pigment Cell Melanoma Res. 2009 Dec;22(6):740-9.
17. Frank NY, Margaryan A, Huang Y, Schatton T, Waaga-Gasser AM, Gasser M, et al. ABCB5-mediated doxorubicin transport and chemoresistance in human malignant melanoma. Cancer Res. 2005 May 15;65(10):4320-33.
18. Siddik ZH. Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene. 2003 Oct 20;22(47):7265-79.
19. Huang Y, Anderle P, Bussey KJ, Barbacioru C, Shankavaram U, Dai Z, et al. Membrane transporters and channels: role of the transportome in cancer chemosensitivity and chemoresistance. Cancer Res. 2004 Jun 15;64(12):4294-301.
20. Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer. 2002 Jan;2(1):48-58.
21. Lockhart AC, Tirona RG, Kim RB. Pharmacogenetics of ATP-binding cassette transporters in cancer and chemotherapy. Mol Cancer Ther. 2003 Jul;2(7):685-98.
22. Boyd AS, Wu H, Shyr Y. Monster cells in malignant melanoma. Am J Dermatopathol. 2005 Jun;27(3):208-10.
23. Brown J. Effects of 2-deoxyglucose on carbohydrate metablism: review of the literature and studies in the rat. Metabolism. 1962 Oct;11:1098-112.
24. Lynch RM, Fogarty KE, Fay FS. Modulation of hexokinase association with mitochondria analyzed with quantitative three-dimensional confocal microscopy. J Cell Biol. 1991 Feb;112(3):385-95.
25. Liu H, Hu YP, Savaraj N, Priebe W, Lampidis TJ. Hypersensitization of tumor cells to glycolytic inhibitors. Biochemistry. 2001 May 8;40(18):5542-7.
26. Liu H, Savaraj N, Priebe W, Lampidis TJ. Hypoxia increases tumor cell sensitivity to glycolytic inhibitors: a strategy for solid tumor therapy (Model C). Biochem Pharmacol. 2002 Dec 15;64(12):1745-51.
27. Maher JC, Krishan A, Lampidis TJ. Greater cell cycle inhibition and cytotoxicity induced by 2-deoxy-D-glucose in tumor cells treated under hypoxic vs aerobic conditions. Cancer Chemother Pharmacol. 2004 Feb;53(2):116-22.
28. Borst P, Evers R, Kool M, Wijnholds J. A family of drug transporters: the multidrug resistance-associated proteins. J Natl Cancer Inst. 2000 Aug 16;92(16):1295-302.
29. Cheung ST, Cheung PF, Cheng CK, Wong NC, Fan ST. Granulin-epithelin precursor and ATP-dependent binding cassette (ABC)B5 regulate liver cancer cell chemoresistance. Gastroenterology. 2011 Jan;140(1):344-55.
30. Mootha VK, Lindgren CM, Eriksson KF, Subramanian A, Sihag S, Lehar J, et al. PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet. 2003 Jul;34(3):267-73.
31. Gazzaniga P, Cigna E, Panasiti V, Devirgiliis V, Bottoni U, Vincenzi B, et al. CD133 and ABCB5 as stem cell markers on sentinel lymph node from melanoma patients. Eur J Surg Oncol. 2010 Dec;36(12):1211-4.
32. Zabierowski SE, Herlyn M. Learning the ABCs of melanoma-initiating cells. Cancer Cell. 2008 Mar;13(3):185-7.
33. Maschek G, Savaraj N, Priebe W, Braunschweiger P, Hamilton K, Tidmarsh GF, et al. 2-deoxy-D-glucose increases the efficacy of adriamycin and paclitaxel in human osteosarcoma and non-small cell lung cancers in vivo. Cancer Res. 2004 Jan 1;64(1):31-4.
34. Goldberg L, Israeli R, Kloog Y. FTS and 2-DG induce pancreatic cancer cell death and tumor shrinkage in mice. Cell Death Dis. 2012;3:e284.