2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/644563摘要：肺癌是發生率相當高的癌症之ㄧ，也是目前台灣癌症死因的第一位。雖然有許多的研究致力於肺癌的治療，肺癌的預後仍然令人失望。肺癌的細胞可以藉由許多的機制產生抗藥性，即使一開始對化療有反應也會很快的復發。癌幹細胞（cancer stem cells）或者腫瘤創始細胞(tunor-initiating cells)的理論提供我們對於癌細胞的起源、抗藥性的產生及預防的方式一個新的研究途徑。從細胞株或臨床檢體中分離或辨認腫瘤創始細胞是可以做的到。但是，培養繼代或是維持腫瘤創始細胞”幹細胞特性”仍有困難。在正常組織的幹細胞，週遭的微環境(microenvironment)或合適的環境（niche）對於幹細胞維持不活動，不分化並保留其生長、分化的能力是非常重要。因此對於腫瘤創始細胞，其週遭的環境也應扮演維持其幹細胞特性的角色。在我們之前的研究，已建立一株特別的細胞株，符合癌幹細胞/腫瘤創始細胞的特性，這株細胞藉由與腫瘤伴生的成纖維細胞(cancer associated fibroblasts)或基質細胞(stromacells)共同培養形成球狀的聚集來維持其幹細胞特性。一但失去其支持細胞（CAFs或stroma cells）它幹細胞的特性便會消失或減少。在其他肺癌細胞株含有較高比例的邊緣族群(sidepopulation)也可以看到球狀的聚集，當這些細胞與CAFs或stroma cells同時培養時。這些球狀聚集的癌細胞有較高幹細胞標誌的表現，例如：nanog，klf-4，Sox-2，Oct4和c-myc基因。我們對於其他具有幹細胞特性的癌細胞是否有同樣的表現感到興趣。癌幹細胞與週遭環境是怎樣互動？在本研究，我們首先會分離癌幹細胞，利用邊緣族群，表面標記，例如：CD133，ALDH，也會利用肺癌病人腫瘤組織做不加血清的原代培養，培養球狀聚集的癌細胞。接著培養CAFs或stoma cells，再將癌幹細胞加入。分析癌細胞的”幹細胞特性”是否仍然維持，並比較共同培養前後癌細胞，CAFs或stoma cells基因表現的差異。CAFs或stoma cells的角色將是重點之ㄧ。我們希望這個研究能闡明癌幹細胞與其周遭微環境的互動，這將提供我們對於癌症藥物治療一個新的對策。<br> Abstract: Lung cancer ranks among the most commonly occurring malignancies and currently is theleading cause of cancer-related death worldwide and in Taiwan. Although a lot of researchfocus on the treatment of lung cancer, the prognosis of lung cancer remains dismal . Lungcancer cells can acquire resistance to chemotherapy by several mechanisms and recur afteran initial response to chemotherapy rapidly. Cancer stem cells (CSCs) or tumor-initiatingcells hypothesis provides us with a new approach to the understanding of carcinogenesis,drug-resistance, and prevention strategies. It is possible to isolate or identify cancer stemcell/ tumor –initiating cells from cancer cell lines or clinical specimens by different methods.However, subculture and maintenance of “stemness” in these cells remained difficult. Innormal stem cells, their microenvironment or “niche” is important to maintain theirquiescent and undifferentiated state, while maintaining their proliferati on and differentiationpotentials. It is suggested CSCs niche to have similar role in maintaining CSCscharacteristics like stem cells. In our preliminary study, we had established a specific cancercell lines with characteristics of CSCs/tumor initiating cells. The cells maintained it sstemness when co-cultured with tumor-associated fibroblasts (or stroma cells) as sphere-likecolonies, and loss characters of CSCs in feeder-free condition. We also found that othercancer cells with higher percentage of side population could form sphere -like colonies whenco-cultured with stroma cells from lung cancer patients. These colonies had high stem cellmarkers such as nanog, klf-4, Sox-2, Oct4 and c-myc. We are interesting that if all cancercells with “stemness” could form sphere-like colonies when co-culture with stroma cells,what will be the interacted mechanisms in the micro environment? In this study, we willisolate cancer stem cells by side population or surface markers such as CD133, ALDH. Wewill also use primary culture without serum to cultivate sphere -like cancer cells. Thenco-culture these cells with available stroma cells or cancer associated fibroblast.Characteristic of the CSCs will be examined; stemness gene expression change in the cancercells and the roles of stroma cells will also be identified. We hope this study could clarify theinteraction between microenvironment and cancer stem cells. It will also provide a newstrategy of drug development for treatment.癌幹細胞微環境肺癌腫瘤伴生的成纖維細胞cancer stem cellsmicroenvironmentlung cancercancer associated fibroblast