臺灣博碩士論文加值系統

癌症為目前世界各國主要死因之一，如何有效對抗癌症已成為現今人類所共同努力之目標。目前雖然有許多方法可以對抗癌細胞，但仍然無法根治，而癌細胞的轉移是治療失敗的主要原因，造成癌細胞轉移的原因有很多，但最主要的是來自於微環境的影響。
腫瘤微環境內充滿各種生長因子，癌細胞與基質細胞間的交互作用對於癌細胞的移動能力具有相當大的影響，它們藉由旁分泌作用進而影響癌細胞的增生或是移動。目前也知道在腫瘤生長期間，組成微環境的基質細胞族群會有所變化，所以基質細胞和癌細胞的交互作用是有次序性的。因此為了研究在腫瘤不同生長時期中癌細胞與基質細胞的交互作用，我們利用三腔體微流道細胞培養晶片，藉以研究纖維母細胞、巨噬細胞對肺腺癌細胞的影響。經由控制不同基質細胞條件式培養液的組合與流動方向，得以模擬在腫瘤生長的不同時期中，參與微環境內的基質細胞族群變化對於癌細胞移動能力的影響。
實驗結果顯示，被癌細胞活化的肌纖維母細胞的條件式培養液會促進癌細胞的移動能力，但巨噬細胞所分泌的TNF-?悜Y先作用到肌纖維母細胞上，則會抑制肌纖維母細胞自分泌TGF-??〞滲鄐O，造成肌纖維母細胞??SMA的表現量與細胞形狀的最小長寬比降低，導致其活性下降，所以肌纖維母細胞促進癌細胞移動的能力也會被減弱。但是當肌纖維母細胞與巨噬細胞所分泌的因子共同作用在癌細胞上時，卻會對癌細胞產生協同作用，使癌細胞的移動能力大幅提升。
因此腫瘤微環境中巨噬細胞扮演非常重要的角色，在腫瘤不同的生長時期中巨噬細胞對癌細胞與其他基質細胞間的交互作用可說是十分重要且複雜的。

In the tumor microenvironment, a lot of stromal cells secret growth factors to enhance the metastasis, proliferation, and survival of cancer cells. A number of cells including fibroblasts and macrophages are involved in the metastasis of a primary tumour. We develop microfluidic cell culture chips that contain three culture chambers to co-culture lung cancer cells, macrophages, and fibroblasts to study the interactions
amount them.
The results showed that cancer cells were activated fibroblasts into myofibroblasts by conditional medium of cancer cell. The myofibroblast conditional medium will promote the mobility of cancer cells, but macrophages secreted TNF-? will inhibit the myofibroblastic ability of autocrine TGF-?? The ??SMA fluorescence intensity and aspect ratio both decreased, so myofibroblasts to promote cancer cell the ability of migration speed will be reduced. But myofibroblasts and macrophages conditional medium together effect on the cancer cells, that will synergies enhance the metastasis of cancer cells. We find that the effect of myofibroblast to increase the migration speed of cancer cells is suppressed by the conditioned medium of the macrophage.
Therefore, macrophages interactions between stromal cells in the tumor micro-environment play a very important role in tumor growth and
metastasis.

摘要 I
Abstract III
目錄 IV
圖表目錄 VI
第一章 序論 1
1-1 癌細胞與轉移 1
1-2 腫瘤微環境 3
1-2-1 纖維母細胞與癌細胞 5
1-2-2 巨噬細胞與癌細胞 7
1-3 研究動機與目的 10
第二章 晶片設計與製作 13
2-1 設計動機 13
2-2 晶片優點 15
2-3 晶片製作 17
2-3-1 PDMS特性 18
2-3-2母模製作 19
2-3-3 PDMS翻模與接合 22
2-4 氣動微閥門 26
2-5 晶片流場分布 28
第三章 實驗方法 30
3-1 細胞培養 30
3-2 系統架設 31
3-3 實驗方法 33
3-4 影像數據分析 42
第四章 結果與討論 44
4-1 癌細胞增生能力 44
4-2 癌細胞移動速率變化 45
4-2-1 實驗A-pretreatment結果 45
4-2-2 實驗B-mixing結果 47
4-2-3 實驗A以及實驗B綜合比較 49
4-3 巨噬細胞抑制myoMRC-5的活性 51
4-3-1 myoMRC-5的最小長寬比變化 51
4-3-2 myoMRC-5的TGF-β1產量 54
4-3-3 巨噬細胞如何抑制myoMRC-5活性 56
4-4 中和TNF-α後癌細胞移動速度變化 62
4-5 結論與未來展望 65
參考文獻 67

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