肺癌在已開發國家中是常見的癌症之一，在台灣，肺癌已經蟬聯十一年癌症死亡率之首。以往罹患肺癌常見的原因為抽菸，但近年來也發現肺癌跟基因遺傳及空氣汙染有關。治療肺癌常見的方法有手術切除、化學治療、放射線治療及標靶藥物治療，儘管現今醫學的進步延長了肺癌病患的存活率，但肺癌細胞的轉移及抗藥性還是現今的一大難題。在本研究中，我們比較了肺腺癌的親代細胞及其抗藥細胞的基因表現量，發現BMP4的表現量在抗藥細胞中高於親代細胞，而且跟上皮細胞間質轉化有關。BMP4也被發現在近端支氣管發育過程中，與胚胎幹細胞轉錄因子SOX2有交互調控的現象。我們在抗藥細胞中敲低BMP4發現會降低抗藥細胞的增生與存活，而在親代細胞中過表達BMP4則發現在低濃度胎牛血清培養的情況下，可以增強親代細胞的存活能力。在添加組蛋白去乙醯酶抑制物的實驗中，我們發現BMP4會藉由表觀遺傳的調控在抗藥細胞中表達，並且我們進一步在親代細胞中敲低HDAC1會發現BMP4的表現量會上升，再次證明BMP4參與表觀遺傳調控。另外，在親代細胞中添加TGF-β的實驗發現TGF-β可以抑制SOX2的表達並促進BMP4表現，進而增強親代細胞的抗藥能力。總體而言，我們發現BMP4在肺腺癌中會促進細胞的存活能力，並證明其參與表觀遺傳調控。

Activating mutation in epidermal growth factor receptor (EGFR) is one of the most common driver mutations in lung adenocarcinoma to initiate deregulated growth, while EGFR-tyrosine kinase inhibitors (EGFR-TKIs) specifically target this mutation exhibit beneficial effect to patients. Although EGFR-TKI brings the initial tremendous response to patients, resistance inevitably occur in 1-2 year. How differentiation signaling of lung branching regulates EGFR-TKI tolerance is unclear. Here, I report that BMP4, a cytokine involved in lung branching, is much more highly enriched in EGFR-TKI tolerant lung cancer cells than their sensitive parental cells, and its overexpression endows cancer cells with high survival ability under low serum stress. We discovered that BMP4 is essential for growth of EGFR-TKI tolerant cells and its silencing can attenuate growth of EGFR-TKI tolerant cells. We observed that TGF-β stimulation can induce BMP4 expression and enhance EGFR-TKI tolerance in lung cancer cells, accompanied with epithelial-to-mesenchymal transition (EMT) and SOX2 downregulation. Meanwhile, we found that romidepsin, a HDAC1/2 specific inhibitor, can induce BMP4 and EMT feature, while knockdown of HDAC1 induces BMP4 expression. These data support the conclusion that BMP4 is under epigenetic control. Together, our findings suggest a critical role of BMP4 in cancer cell survival and indicate a potential of BMP4 as a therapeutic target in lung cancer treatment.

Abstract--------------- 1
中文摘要- --------------- 2
致謝------------------- 3
Introduction----------- 6
Materials and Methods-- 11
Results---------------- 16
Summary---------------- 21
Figures---------------- 23
References------------- 37

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