臺灣博碩士論文加值系統

乳腺癌是國人女性惡性腫瘤中的第四名並且在全球的發生率持續增加。其中三陰性乳腺癌(Triple-negative breast cancer,TNBC)佔所有乳腺癌約10-15%，被定義為缺乏雌激素受體(Estrogen receptor,ER)、黃體激素受體(Progesterone receptor ,PR)及人類上皮生長因子受體2(Human epidermal growth factor receptor2,HER2)三陰性乳癌因無特定受體表現。不適合使用賀爾蒙治療及標靶治療，目前尚未有較特定之臨床治療。過去的文獻指出，Y-box binding protein1(YB-1)藉由轉錄和轉譯機制來調控基因表現，並參與腫瘤生長和調控HER2和ER之表現。在此篇研究中探討2-phenylnaphthyridin-4-ones(2-PN)的衍生物CSC-3436是否可藉由抑制YB-1使ERα重新表現並增加Tamoxifen對三陰性乳癌細胞之毒殺作用。，研究結果顯示，三陰性乳癌細胞中CSC-3436藉由抑制YB-1的表現，進而造成ERα表現量恢復並增強Tamoxifen的抗癌作用及促進細胞凋亡。進一步發現CSC-3436可透過Akt路徑的調控來抑制TWIST及下游YB-1的表現。因此CSC-3436協同Tamoxifen能誘導細胞凋亡，並增加對三陰性乳癌細胞的抗癌作用，在TWIST以及其下游YB-1也能提供相關的分子標靶治療。

Breast cancer is the most common malignancy among women in Taiwan and its incidence is increasing worldwide. Triple-negative breast cancer (TNBC) accounts for approximately 20% of breast cancer, it is defined by a lack of expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). However, TNBC is a non-specific receptor expression and is therefore not suitable for use hormone therapy and target therapy, currently there is no specific clinical treatment. Y-box binding protein 1 (YB-1) controls gene expression through both transcriptional and translational mechanisms and is involved in various biological activities such as tumor progression. Expression of HER2 and ER depends upon YB-1 in human breast cancers. In this study, we used (2-PN) CSC-3436, to address a the derivatives hypothesis of that 2-phenylnaphthyridin-4-ones CSC-3436 could inhibit YB-1 expression and re-express ER-α in TNBC cells. The results showed that CSC-3436 decreased YB-1 expression in TNBC cells. CSC-3436 also increased ER-α expression and enhanced the anticancer effect of tamoxifen through inducing apoptosis in TNBC cells. We next found that CSC-3436 inhibited YB-1 expression through decreased Twist expression and Akt signal pathway. Our analysis indicated that Twist and its downstream effector YB-1 provide molecular targets for CSC-3436 synergized with tamoxifen against TNBC cells through the induction of apoptosis.

致謝 ...............I
中文摘要 ...............III
英文摘要 .................. IV
目次 ................V
圖表目次 ...............VIII
縮寫表 ...............X
第一章 前言 ...............1
第一節 癌症 (Cancer) ...............1
第二節 乳癌 (Breast cancer) ...............4
第三節 三陰性乳癌 (Triple-negative breast cancer) ...............7
第四節 細胞凋亡 (Apoptosis) ...............10
第五節 Y-box 結合蛋白 (Y-box binding protein, YB-1）...............13
第六節 CSC-3436 化合物介紹 ...............16
第七節 研究動機 ...............19
第二章 材料與方法...............20
第一節 材料來源 ...............20
第二節 實驗方法...............25
第三章 研究結果...............37
第一節 CSC-3436 對三陰性乳癌細胞株 ERα 和 YB-1 蛋白質表現量之影響 ..... 37
1.1 CSC-3436 對三陰性乳癌細胞株的 YB-1 蛋白表現量之影響 ..... 37
1.2 抑制 YB-1 蛋白質表現對 MDA-MB-231 細胞株 ERα 表現之影響...............39
1.3 CSC-3436 協同 Tamoxifen 對於 MDA-MB-231 細胞株之存活率(Cell viability)影響... 41
1.4 CSC-3436 協同 Tamoxifen 對於 MDA-MB-231 細胞株之死亡率(Apoptosis)影響..... 43
第二節 CSC-3436 對三陰性乳癌細胞中抑制 YB-1 蛋白質之分子機制...............45
2.1 CSC-3436 對於 YB-1 mRNA 表現之影響 ...............45
2.2 CSC-3436 對於 YB-1 蛋白質穩定度之影響 ...............47
第三節 CSC-3436 對三陰性乳癌細胞中 DNMT1 蛋白質表現之影響...............49
第四節 CSC-3436 在 MDA-MB-231 細胞株中是否透過 YB-1 上游路徑調控 ERα 之表現 ...............51
4.1 CSC-3436 對於 YB-1 上游調控因子 Twist 和 Slug 之蛋白質表現影響 ...............51
4.2 抑制 Twist 蛋白質表現對 MDA-MB-231 細胞株 YB-1 表現之影響...............53
4.3 CSC-3436 在 MDA-MB-231 細胞株對 Twist 上游相關路徑之影響...............55
第四章 討論 ...............57
第五章 結論 ...............59
第六章 參考文獻...............63

Negrini, S., V.G. Gorgoulis, and T.D. Halazonetis, Genomic instability--an evolving hallmark of cancer. Nat Rev Mol Cell Biol, 2010. 11(3): p. 220-8.

Weiderpass, E., Lifestyle and cancer risk. J Prev Med Public Health, 2010. 43(6): p. 459-71.

.

Hanahan, D. and R.A. Weinberg, Hallmarks of cancer: the next generation. Cell, 2011. 144(5): p. 646-74.

Lalloo, F. and D.G. Evans, Familial breast cancer. Clin Genet, 2012. 82(2): p. 105-14.

Chu, Q.D., et al., High chemokine receptor CXCR4 level in triple negative breast cancer specimens predicts poor clinical outcome. J Surg Res, 2010. 159(2): p. 689-95.

.

Sorlie, T., et al., Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci U S A, 2001. 98(19): p. 10869-74.

.

Dent, R., et al., Pattern of metastatic spread in triple-negative breast cancer. Breast Cancer Res Treat, 2009. 115(2): p. 423-8.

Anders, C.K. and L.A. Carey, Biology, metastatic patterns, and treatment of patients with triple-negative breast cancer. Clin Breast Cancer, 2009. 9 Suppl 2: p. S73-81.

Igney, F.H. and P.H. Krammer, Death and anti-death: tumour resistance to apoptosis. Nat Rev Cancer, 2002. 2(4): p. 277-88.

Elmore, S., Apoptosis: a review of programmed cell death. Toxicol Pathol, 2007. 35(4): p. 495-516.

Haupt, Y. and M. Oren, p53-mediated apoptosis: mechanisms and regulation. Behring Inst Mitt, 1996(97): p. 32-59.

Haupt, Y., et al., p53 mediated apoptosis in HeLa cells: transcription dependent and independent mechanisms. Leukemia, 1997. 11 Suppl 3: p. 337-9.

Chowdhury, D., et al., The exonuclease TREX1 is in the SET complex and acts in concert with NM23-H1 to degrade DNA during granzyme A-mediated cell death. Mol Cell, 2006. 23(1): p.
133-42.

Eliseeva, I.A., et al., Y-box-binding protein 1 (YB-1) and its functions. Biochemistry (Mosc), 2011. 76(13): p. 1402-33.

Kohno, K., et al., The pleiotropic functions of the Y-box-binding protein, YB-1. Bioessays, 2003. 25(7): p. 691-8.

Stratford, A.L., et al., Targeting p90 ribosomal S6 kinase eliminates tumor-initiating cells by inactivating Y-box binding protein-1 in triple-negative breast cancers. Stem Cells, 2012. 30(7): p.1338-48.

Bader, A.G. and P.K. Vogt, Phosphorylation by Akt disables the anti-oncogenic activity of YB-1. Oncogene, 2008. 27(8): p. 1179-82.

Basaki, Y., et al., Akt-dependent nuclear localization of Y-box-binding protein 1 in acquisition of malignant characteristics by human ovarian cancer cells. Oncogene, 2007. 26(19): p.2736-46.