請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74165
標題: | 間充質幹細胞衍生之腫瘤微環境介導MAPK信號途徑中c-Fos S374的磷酸化並促進肺癌進程 Mesenchymal Stem Cells-derived Microenvironment Promotes Lung Cancer Cell Progression via c-Fos S374 Phosphorylation in MAPK Signaling Pathway |
作者: | Chia-Chi Wang 王嘉琪 |
指導教授: | 阮雪芬 |
關鍵字: | 腫瘤微環境,間充質幹細胞,磷酸化蛋白質體學,絲裂原活化蛋白激?信號途徑,轉錄因子c-Fos,異位ATP合成?,P2X7受體,嘌呤能信號傳導途徑, tumor microenvironment,mesenchymal stem cells (MSCs),phosphoproteomics,mitogen-activated protein kinases (MAPK) signaling pathway,c-Fos,ectopic ATP synthase,P2X7 receptor,purinergic signaling, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 腫瘤微環境由腫瘤細胞與異質性的基質細胞組合而成,包括間充質幹細胞與其衍生的成纖維細胞。然而這些基質細胞能夠透過旁分泌趨化因子及生長因子來調節腫瘤的生成,血管新生以及促進癌症轉移及侵襲。在本研究中我們試圖探討間充質幹細胞在肺癌腫瘤微環境中所扮演的角色。首先我們以培養間充質幹細胞三天的條件培養基對於肺癌細胞進行處理,結果發現間充質幹細胞的條件培養基會增強肺癌細胞的生長能力及遷移情形。進一步我們想了解其中的調控機制,我們以磷酸化蛋白質體學的方式剖析其參與調控的訊號途徑,我們總共鑑定到有差異表現的磷酸化蛋白700個,其中含有1926個磷酸化位點,此外我們進行激酶的預測分析,發現間充質幹細胞透過激活肺癌細胞中絲裂原活化蛋白激酶信號途徑 (MAPK pathway) 促使轉錄因子c-Fos在磷酸化位點絲氨酸374處 (serine 374) 的活化。進一步我們透過使用MAPK抑制劑來抑制c-Fos S374的磷酸化,結果發現原本肺癌細胞增強的生長及遷移能力因此而受到抑制。另一方面我們也構築了去磷酸化的c-Fos質體DNA,將其於細胞中過度表現。在與野生型的c-Fos相比之下,去磷酸化的c-Fos導致細胞增殖與集落形成的能力下降,而細胞的遷移情形也受到抑制。整合上述實驗結果,我們的研究表明,間充質幹細胞所衍生的腫瘤微環境會介導肺癌細胞MAPK信號途徑的活化,進一步將c-Fos S374磷酸化,並且促進肺癌細胞的生長及轉移。
另一方面,根據最近的文獻報導顯示腫瘤微環境中存在著大量的三磷酸腺苷(ATP),其能透過嘌呤能信號傳導途徑活化癌細胞膜上的P2X7受體。然而在我們的研究中觀察到間充質幹細胞的細胞表面存在著大量的異位ATP合成酶,此外腫瘤微環境中的ATP會激活肺癌細胞的MAPK信號途徑,進一步導致c-Fos S374的磷酸化。綜合來說,我們的研究證明,間充質幹細胞表面的異位ATP合成酶會釋放大量的ATP進入肺癌的腫瘤微環境,並且透過MAPK信號途徑的活化進一步將c-Fos S374磷酸化,最後促進肺癌細胞的生長以及轉移能力。 Tumor microenvironment contains tumor cells and a mixture of heterogeneous stromal cells, including mesenchymal stem cells (MSCs) and MSC-derived fibroblast. These stromal cells are able to regulate tumor growth, angiogenesis, metastasis and invasion by paracrine secretion of cytokines, chemokine and growth factors. Here, we attempt to dissect the roles of MSCs in the tumor microenvironment of lung cancer. At first, we found that the MSCs conditioned medium (MSC-CM) enhanced lung cancer cell proliferation and migration. However, the signaling pathways mediated by MSC-secreting factors are still unclear. To further elucidate MSC regulatory pathways, we performed quantitative phosphoproteomics for MSC-CM treated lung cancer cells, and a total of 1926 phosphorylation sites on 700 phosphoproteins were identified. Moreover, integrative analysis of phosphoproteins and predicted kinases suggested that phosphorylation of c-Fos at serine 374 was induced by MSC-CM through activating mitogen-activated protein kinases (MAPK) signaling pathway in lung cancer cells. Consistently, our results showed that MSC-induced cell proliferation and migration were abrogated by inhibiting c-Fos phosphorylation using ERK1/2 inhibitor. In addition, the dephosphorylated form of c-Fos at serine 374 led to decreased cell proliferation, cell motility and colony forming ability compared to wild-type form of c-Fos in MSC-CM treated lung cancer cells. Collectively, our study demonstrated that MSCs might trigger the MAPK pathway followed by the phosphorylation of c-Fos at serine 374 to promote cancer proliferation and migration. Recent studies have reported that extracellular ATPs accumulate in the tumor microenvironment and stimulate the P2X7 receptor on the cancer cell membrane through purinergic signaling. In our data, we observed that ectopic ATP synthase was overexpressed on the cell surface of MSCs. In addition, extracellular ATPs in the lung cancer microenvironment trigger the MAPK pathway to phosphorylate c-Fos on serine 374. In conclusion, our study suggested that ectopic ATP synthase on the surface of MSCs released extracellular ATPs into the lung cancer microenvironment and promoted cancer progression through c-Fos S374 phosphorylation in the MAPK signaling pathway. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74165 |
DOI: | 10.6342/NTU201903131 |
全文授權: | 有償授權 |
顯示於系所單位: | 分子與細胞生物學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-108-1.pdf 目前未授權公開取用 | 5.79 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。