三陰性乳癌的特徵是易於生長、轉移，具有高異質性、高復發性及高抗藥性，基因調控錯誤會重塑細胞的命運並誘導腫瘤生長。在此篇論文，我們利用MCF-10A作為研究材料，探討乙型轉化生長因子第三型受體（TGFBR3）在乳腺腺泡形成過程中，對其異質性與細胞生死抉擇產生的影響。TGFBR3幫助接收受質，是TGFβ訊號傳遞途徑中的一員。實驗結果顯示，當細胞脫離細胞外間質，TGFBR3會被大量表現，BCL2相關蛋白的表現量會被調節，最後導致細胞死亡。抑制TGFBR3則可以使細胞壓力相關基因表現量上升，使細胞凋亡的現象受到抑制。顯示TGFBR3的表現能夠影響細胞的生死抉擇。TGFBR3和細胞壓力之間的關係對於細胞命運的影響讓我們從新看待乳腺形成的過程，或許可以提供治療乳癌的新策略。

Triple-negative breast cancer is characterized by high proliferation, metastases, heterogeneity incidence of relapse, and drug resistance. Misregulation of gene expression re-programs cell fate and promotes tumor progression. Here, we target transforming growth factor beta receptor III (TGFBR3) and use MCF-10A as a material to investigate the life-death decision in mammary acinar morphogenesis. The results suggest that TGFBR3 expression is induced while cells detach from extracellular matrix (ECM), which is critical for cell survival. In this stressful situation, epithelial cells undergo apoptosis through BCL2 pathway during ECM detachment. Knockdown of TGFBR3 lead to upregulation of cell stress genes, resulting in apoptosis inhibition. The TGFBR3-stress response genes axis for cell fate determination provides a new insight of mammary gland morphogenesis and might pave the way for new therapeutic strategies.

摘要 i
Abstract ii
謝誌 iii
目錄 iv
圖目錄 vi
第一章 緒論 1
1-1 三陰性乳癌 1
1-2 MCF-10A細胞株 3
1-3 MCF-10A乳腺腺泡的細胞間異質性 9
1-4 乙型轉化生長因子第三型受體（TGFBR3） 11
1-5 上皮間質轉換 13
第二章 實驗材料與方法 15
2-1細胞株 15
2-2質體 15
2-3反轉錄定量聚合酶連鎖反應 （RT-qPCR） 15
2-4西方墨點法（Western blotting） 17
2-5懸浮實驗 18
2-6統計方法 21
第三章 實驗結果 22
3-1 當細胞脫離ECM會造成細胞表現TGFBR3，將促使細胞凋亡 22
3-1-1 結論 22
3-1-2當乳腺腺泡形成時，TGFBR3被大量表現在內層細胞中。 22
3-1-3 TGFBR3被表現在無法貼附ECM的細胞中 24
3-1-4 懸浮培養細胞時，抑制TGFBR3可減緩細胞凋亡 26
3-1-5懸浮培養細胞會影響BCL2相關蛋白的mRNA表現量，抑制TGFBR3可以減緩此情況 29
3-1-6 TGFBR3抑制ATF4表現而導致細胞凋亡 42
3-2 TGFBR3經由SMADs讓乳腺上皮細胞進行上皮間質轉移 46
3-2-1 結論 46
3-2-2 TGFBR3讓細胞趨於表現間質細胞型態，而非上皮細胞型態 47
3-2-3 TGFBR3將TGFβ訊號引流至磷酸化SMAD1／5而非磷酸化SMAD2／3 50
第四章 討論 54
第五章 參考文獻 56

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