Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/99397
標題: Metformin與Emodin對於雄性激素受體蛋白及攝護腺癌LNCaP細胞增殖的協同抑制作用
The synergistic inhibition of Metformin and Emodin on androgen receptor protein and LNCaP cell proliferation
作者: 方晧
Hao Fang
關鍵字: 每福敏;大黃素;雄性激素受體;攝護腺癌;Androgen receptor;Emodin;Metformin;Prostate cancer
引用: 參考文獻 1. Singapore Urological Association Clinical Guidelines for Male Lower Urinary Tract Symptoms/Benign Prostatic Hyperplasia. Singapore Med J, 2017. 58(8): p. 473-480. 2. Castro, E. and R. Eeles, The role of BRCA1 and BRCA2 in prostate cancer. Asian J Androl, 2012. 14(3): p. 409-14. 3. Chodak, G.W., P. Keller, and H.W. Schoenberg, Assessment of screening for prostate cancer using the digital rectal examination. J Urol, 1989. 141(5): p. 1136-8. 4. Pron, G., Prostate-Specific Antigen (PSA)-Based Population Screening for Prostate Cancer: An Evidence-Based Analysis. Ont Health Technol Assess Ser, 2015. 15(10): p. 1-64. 5. Giovacchini, G., et al., (11)C-choline PET/CT predicts survival in prostate cancer patients with PSA < 1 NG/ml. Eur J Nucl Med Mol Imaging, 2019. 6. Hoang, D.T., et al., Androgen receptor-dependent and -independent mechanisms driving prostate cancer progression: Opportunities for therapeutic targeting from multiple angles. Oncotarget, 2017. 8(2): p. 3724-3745. 7. Narayanan, S., S. Srinivas, and D. Feldman, Androgen-glucocorticoid interactions in the era of novel prostate cancer therapy. Nat Rev Urol, 2016. 13(1): p. 47-60. 8. Tan, M.H., et al., Androgen receptor: structure, role in prostate cancer and drug discovery. Acta Pharmacol Sin, 2015. 36(1): p. 3-23. 9. Brand, J.S., et al., Testosterone, sex hormone-binding globulin and the metabolic syndrome in men: an individual participant data meta-analysis of observational studies. PLoS One, 2014. 9(7): p. e100409. 10. Ferrari, N., et al., Adaptive phenotype drives resistance to androgen deprivation therapy in prostate cancer. Cell Commun Signal, 2017. 15(1): p. 51. 11. Beitel, L.K., et al., Mechanisms mediating spinal and bulbar muscular atrophy: investigations into polyglutamine-expanded androgen receptor function and dysfunction. Front Neurol, 2013. 4: p. 53. 12. Russo, J.W., et al., Phosphorylation of androgen receptor serine 81 is associated with its reactivation in castration-resistant prostate cancer. Cancer Lett, 2018. 438: p. 97-104. 13. Hostalek, U., M. Gwilt, and S. Hildemann, Therapeutic Use of Metformin in Prediabetes and Diabetes Prevention. Drugs, 2015. 75(10): p. 1071-94. 14. Sun, J., et al., Deciphering Signaling Pathway Networks to Understand the Molecular Mechanisms of Metformin Action. PLoS Comput Biol, 2015. 11(6): p. e1004202. 15. Viollet, B., et al., Cellular and molecular mechanisms of metformin: an overview. Clin Sci (Lond), 2012. 122(6): p. 253-70. 16. De Santi, M., et al., Metformin prevents cell tumorigenesis through autophagy-related cell death. Scientific Reports, 2019. 9(1): p. 66. 17. Peng, M., N. Yin, and M.O. Li, SZT2 dictates GATOR control of mTORC1 signalling. Nature, 2017. 543(7645): p. 433-437. 18. Kourelis, T.V. and R.D.J.M.O. Siegel, Metformin and cancer: new applications for an old drug. 2012. 29(2): p. 1314-1327. 19. Wang, Y., et al., Metformin represses androgen-dependent and androgen-independent prostate cancers by targeting androgen receptor. Prostate, 2015. 75(11): p. 1187-96. 20. Demir, U., et al., Metformin anti-tumor effect via disruption of the MID1 translational regulator complex and AR downregulation in prostate cancer cells. BMC Cancer, 2014. 14: p. 52. 21. Shrimali, D., et al., Targeted abrogation of diverse signal transduction cascades by emodin for the treatment of inflammatory disorders and cancer. Cancer Lett, 2013. 341(2): p. 139-49. 22. Huang, Q., et al., Emodin inhibits tumor cell adhesion through disruption of the membrane lipid Raft-associated integrin signaling pathway. Cancer Res, 2006. 66(11): p. 5807-15. 23. Li, W.Y., et al., Emodin elicits cytotoxicity in human lung adenocarcinoma A549 cells through inducing apoptosis. Inflammopharmacology, 2014. 22(2): p. 127-34. 24. Kim, J., et al., Emodin suppresses maintenance of stemness by augmenting proteosomal degradation of epidermal growth factor receptor/epidermal growth factor receptor variant III in glioma stem cells. Stem Cells Dev, 2015. 24(3): p. 284-95. 25. Jayasuriya, H., et al., Emodin, a protein tyrosine kinase inhibitor from Polygonum cuspidatum. J Nat Prod, 1992. 55(5): p. 696-8. 26. Huang, P.H., et al., Emodin and Aloe-Emodin Suppress Breast Cancer Cell Proliferation through ER alpha Inhibition. Evid Based Complement Alternat Med, 2013. 2013: p. 376123. 27. Yu, C.X., et al., Emodin induces apoptosis in human prostate cancer cell LNCaP. Asian J Androl, 2008. 10(4): p. 625-34. 28. Cha, T.L., et al., Emodin down-regulates androgen receptor and inhibits prostate cancer cell growth. Cancer Res, 2005. 65(6): p. 2287-95. 29. Dong, X., et al., Emodin: A Review of its Pharmacology, Toxicity and Pharmacokinetics. Phytother Res, 2016. 30(8): p. 1207-18. 30. Tran, L.N.K., et al., The Combination of Metformin and Valproic Acid Has a Greater Anti-tumoral Effect on Prostate Cancer Growth In Vivo than Either Drug Alone. In Vivo, 2019. 33(1): p. 99-108. 31. Bojkova, B., et al., Metformin and melatonin improve histopathological outcome of NMU-induced mammary tumors in rats. Pathol Res Pract, 2019. 32. Ballangrud, A.M., et al., Growth and characterization of LNCaP prostate cancer cell spheroids. Clin Cancer Res, 1999. 5(10 Suppl): p. 3171s-3176s. 33. Malaguarnera, R., et al., Metformin inhibits androgen-induced IGF-IR up-regulation in prostate cancer cells by disrupting membrane-initiated androgen signaling. Endocrinology, 2014. 155(4): p. 1207-21. 34. Zhang, Y., et al., Metformin Ameliorates Uterine Defects in a Rat Model of Polycystic Ovary Syndrome. EBioMedicine, 2017. 18: p. 157-170. 35. Lee, S.Y., et al., SMILE upregulated by metformin inhibits the function of androgen receptor in prostate cancer cells. Cancer Lett, 2014. 354(2): p. 390-7. 36. Jaune, E. and S. Rocchi, Metformin: Focus on Melanoma. Front Endocrinol (Lausanne), 2018. 9: p. 472. 37. Sarmento-Cabral, A., et al., Metformin Reduces Prostate Tumor Growth, in a Diet-Dependent Manner, by Modulating Multiple Signaling Pathways. Mol Cancer Res, 2017. 15(7): p. 862-874. 38. Iyer, V.V., Small molecules for immunomodulation in cancer: a review. Anticancer Agents Med Chem, 2015. 15(4): p. 433-52. 39. Ok, S., et al., Emodin inhibits invasion and migration of prostate and lung cancer cells by downregulating the expression of chemokine receptor CXCR4. Immunopharmacol Immunotoxicol, 2012. 34(5): p. 768-78. 40. Drewinko, B., et al., Combination chemotherapy in vitro with adriamycin. Observations of additive, antagonistic, and synergistic effects when used in two-drug combinations on cultured human lymphoma cells. Cancer Biochem Biophys, 1976. 1(4): p. 187-95. 41. Gao, W., C.E. Bohl, and J.T. Dalton, Chemistry and structural biology of androgen receptor. Chem Rev, 2005. 105(9): p. 3352-70. 42. Cha, T.L., et al., Emodin modulates epigenetic modifications and suppresses bladder carcinoma cell growth. Mol Carcinog, 2015. 54(3): p. 167-77. 43. Chien, S.W., et al., Growth Modulation of Diabetic Factors and Antidiabetic Drugs on Prostate Cancer Cell Lines. Chin J Physiol, 2016. 59(2): p. 109-18. 44. Peng, M., et al., Combination of metformin with chemotherapeutic drugs via different molecular mechanisms. Cancer Treat Rev, 2017. 54: p. 24-33. 45. Mayer, M.J., L.H. Klotz, and V. Venkateswaran, Metformin and prostate cancer stem cells: a novel therapeutic target. Prostate Cancer Prostatic Dis, 2015. 18(4): p. 303-9. 46. Hsu, F.N., et al., Regulation of androgen receptor and prostate cancer growth by cyclin-dependent kinase 5. J Biol Chem, 2011. 286(38): p. 33141-9. 47. Mijatovic, S., et al., Naturally occurring compounds in differentiation based therapy of cancer. Biotechnol Adv, 2018. 36(6): p. 1622-1632. 48. Lin, W.F., C. Wang, and C.Q. Ling, [Research progress in anti-tumor effect of emodin]. Zhongguo Zhong Yao Za Zhi, 2015. 40(20): p. 3937-40. 49. Zu, C., et al., Low dose Emodin induces tumor senescence for boosting breast cancer chemotherapy via silencing NRARP. Biochem Biophys Res Commun, 2018. 505(4): p. 973-978. 50. He, L. and F.E. Wondisford, Metformin action: concentrations matter. Cell Metab, 2015. 21(2): p. 159-162. 51. Honjo, S., et al., Metformin sensitizes chemotherapy by targeting cancer stem cells and the mTOR pathway in esophageal cancer. Int J Oncol, 2014. 45(2): p. 567-74. 52. Zhuang, Y., et al., Mechanisms by which low glucose enhances the cytotoxicity of metformin to cancer cells both in vitro and in vivo. PLoS One, 2014. 9(9): p. e108444. 53. Biernacka, K.M., et al., Hyperglycaemia-induced resistance to Docetaxel is negated by metformin: a role for IGFBP-2. Endocr Relat Cancer, 2016. 24(1): p. 17-30. 54. Su, Y.J., et al., Role of Rad51 down-regulation and extracellular signal-regulated kinases 1 and 2 inactivation in emodin and mitomycin C-induced synergistic cytotoxicity in human non-small-cell lung cancer cells. Mol Pharmacol, 2010. 77(4): p. 633-43. 55. Kim, Y.S., et al., Emodin Sensitizes Hepatocellular Carcinoma Cells to the Anti-Cancer Effect of Sorafenib through Suppression of Cholesterol Metabolism. Int J Mol Sci, 2018. 19(10).
摘要: 
攝護腺癌全世界中,具有相當高的發生率以及死亡率,而在台灣攝護腺癌的發生率逐年上升且居高不下的死亡率。目前許多研究指出,攝護腺癌的發生及惡化與雄性激素和雄性激素受體(Androgen receptor, AR)的表現量有密切的關係;甚至有研究表示,AR也可加速雄激素非依賴性攝護腺癌的癌細胞生長,也與攝護腺癌復發和轉移特性有關。因此抑制雄性激素與AR是治療攝護腺癌的方式之一。本實驗室在過去發表的文獻中提到,AR Ser 81位點磷酸化對於AR蛋白質的穩定性相當重要。Metformin主要用於治療糖尿病的藥物,可經由抑制攝護腺癌重要生長指標AR mRNA的表現進而影響AR的訊息傳遞路徑。許多研究發現,Metformin可透過多種路徑達到抑癌效果,包括抑制致癌基因(Oncogene)、血管生成和促生長因子;然而,在現今癌症研究顯示,Metformin在癌症治療的效應,包括與其他抗癌藥物合併治療的影響甚至在癌細胞中的作用機轉還有很多部分是未知的。此外,Emodin是一種在中草藥中常見的成分,具有抗發炎,抗氧化和肝保護功能,其抗癌活性在近期也有廣泛的研究,且Emodin可抑制多種癌細胞生長和與促使其凋亡,並且通過誘導AR降解而抑制攝護腺癌細胞生長。因此,本篇論文想探討Metformin與Emodin兩者對AR有不同抑制作用的藥物,在合併處理下,對於抑制AR蛋白穩定性、AR mRNA表現及協同抑制攝護腺癌細胞株生長的情形。研究結果發現,在Metformin 8 mM與Emodin 10 µM合併處理組發現抑制AR蛋白量的能力明顯增強,在量化統計分析也有顯著差異;而以Emodin 10µM作為對照組,在Metformin與Emodin合併處理發現細胞數量隨著Metformin的增加而隨之下降,在Metformin 4、8 mM兩組具有顯著差異,在Metformin 8 mM與Emodin 10 µM合併處理組有最高CI (Combination index)值2.537,代表在其濃度處理下具有協同作用。在AR降解方面,Metformin 8 mM與Emodin 10 µM處理下會使LNCaP細胞株中的AR蛋白質降解速度上升。在基因表現方面,Metformin 2、4、8 mM搭配Emodin 10 µM處理下與對照組比較經統計分析有顯著差異,得知Emodin對於Metformin抑制AR mRNA的表現具有加強作用。依上述結果得知,在兩者藥物合併處理下,對於LNCaP生長顯著抑制且具有協同作用;合併處理可以減少Metformin與Emodin的劑量和副作用。期許透過本篇研究的發現,能夠對於未來攝護腺癌治療方面有所貢獻。

Prostate cancer has a high incidence and mortality rate in the world. At present, many studies have shown that the occurrence and deterioration of prostate cancer are closely related to androgen receptor (AR). It is known that AR increases the growth, recurrence and metastasis of androgen-independent prostate cancer. Therefore, inhibition of AR can be one of the ways to treat prostate cancer. Our group has published that AR Ser81 site phosphorylation is important for the stability of AR protein. In addition, Metformin which is a clinical drug for diabetes patients has the anti-cancer ability including inhibitions of oncogenic ability, angiogenesis, and growth factor effects. Interestingly, Metformin was found to inhibit AR-related signaling by decreasing AR expression. On the other hand, Emodin, a compound in Chinese herbal medicine, has anti-inflammatory, anti-oxidant, and liver protective functions. Its anti-cancer activity has also recently been extensively studied such as the inhibitions of cancer growth and induction of apoptosis. Besides, Emodin can inhibit prostate cancer cell growth by degrading AR protein. Therefore, the aim of this study is to explore the effects of the combination treatment of Metformin and Emodin on prostate cancer cell line, LNCaP, and investigate whether AR protein degradation is the target of the treatment. After identifying the individual efficiencies of Metformin and emodin to AR protein decrease in LNCaP cells, we found the further decrease of AR protein level with the combination treatment and the AR decline was also identified as proteasome-dependent degradation. In this study, it was found that the combination of Metformin 8 mM and Emodin 10 µM showed that the ability to significantly inhibit AR protein level, and there were significant differences by quantitative statistical analysis. Comparing the combination treatment group Metformin 8mM and Emodin 10 µM to control group Emodin 10 µM, that the cell number was decreased following by increasing concentration of Metformin. In addition, Metformin 4 mM and 8 mM had significant differences. The Metformin 8 mM and Emodin 10 µM combined treatment group had the highest Combination Index value (CI=2.537), representing a synergistic effect under this combination. In terms of AR degradation, combined treatment with Metformin 8 mM and Emodin 10 µM could accelerate AR protein degradation in LNCaP cell line. In terms of gene expression, there was also a statistical difference between combination treatment and with the control group. It is known that Metformin can enhance the effect of Emodin in inhibiting AR mRNA. At last, LNCaP cell growth was significantly inhibited by combination treatment of Metformin and Emodin in a synergistic manner. We hope these findings might contribute to the future treatment for prostate cancer by using the strategy of combination treatment of Metformin and Emodin.
URI: http://hdl.handle.net/11455/99397
Rights: 同意授權瀏覽/列印電子全文服務,2022-02-15起公開。
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