探討間葉幹細胞與非小細胞肺癌細胞經自發性融合後產生融合細胞之特性

Characterization of fusion cells derived from spontaneous fusion events between mesenchymal stem cells and non-small cell lung cancer cells

柯承佑(Cherng-You Ke)

指導教授 ： 鄧文炳

共同指導教授 ： 吳駿翃

臺北醫學大學/口腔醫學院/生醫材料暨工程研究所/碩士(2011年)

摘要

細胞融合是一連串過程複雜的細胞間交互作用，其對於體內組織的發展以及平衡是相當重要的，例如肌肉纖維由肌母細胞融合而形成以及免疫反應中巨噬細胞由單核球融合而成等。最近的研究結果顯示，間葉幹細胞可以藉由與細胞進行融合反應修復受損的組織。此外癌細胞亦被證實在其生長的過程當中透過不斷的分泌細胞激素以及趨化激素，而讓受吸引的間葉幹細胞認為癌症病灶處為一個”未癒合的傷口”。許多種癌細胞會任意的和癌細胞本身或和體細胞進行融合，而這些融合會使癌細胞更加惡性，例如增加細胞分裂能力、轉移性和抗藥性。因此我們假設，癌細胞會經由與幹細胞的融合而提高其惡性。本研究中，我們將帶有紅螢光蛋白的間葉幹細胞與帶有綠螢光蛋白的非小細胞肺癌細胞進行共同培養，一方面利用兩種抗生素來篩選，另一方面以兩種螢光表現來觀察融合成功的細胞。從本研究的初步結果可以看出，融合的細胞在側群細胞的比例上明顯的高於親代的非小細胞肺癌細胞。再跟親代的肺癌細胞做型態上的比對，融合細胞在型態上與肺癌細胞的上皮細胞型態已經不相同，其型態變成趨於紡錘狀的間葉細胞型態，而且在體外的細胞遷移實驗上，融合細胞也表現出了優於肺癌細胞的移動能力。總而言之，本研究中我們證實了在經過間葉幹細胞以及非小細胞肺癌細胞自發性的融合之後，融合細胞的抗藥性以及細胞的移動能力上都有顯著的提升，最後期望了解間葉幹細胞在癌症發展上角色以及其機制。

關鍵字

細胞融合 ； 非小細胞肺癌 ； 間葉幹細胞 ； 共同培養 ； 側群細胞

並列摘要

Cell fusion is an intricate cellular event that is critical to tissue development and homeostasis, such as formation of muscle fibers (fusion of myoblasts) or immune response formation of macrophage). Recent studies have indicated that mesenchymal stem cells (MSCs) can regenerate damaged tissue via cell fusion. Moreover, cancer cells can attract MSCs due to the constant deposition of cytokines and chemokines that renders itself as a “wound that never heals”. Many cancer cell types fuse promiscuously with themselves or somatic cells to enhance malignancy, such as higher proliferation ability, metastatic capability and drug resistance. Hence, we hypothesized that tumor cells could gain aggressive malignancy through fusion with MSCs. In this study, we directly co-cultured MSCs containing RFP and non-small cell lung cancer cells (NSCLCs) containing GFP and then identified fused cells with double antibiotics selection and distinct fluorescence. Our preliminary results showed higher percentage of side population in fusion cells than that in parental cancer cells. Comparable to parental cancer cells, the fused cells revealed mesenchymal phenotype and increased motility in in vitro migration assay. In summary, we demonstrated that the fusion cell derived from MSCs and NSCLCs displayed greater drug resistance and cell motility than parental cancer cells. The completion of this study would provide an alternative mechanism by which endogenous MSCs were implicated in cancer development.

並列關鍵字

Cell fusion ； Non-small cell lung cancer ； Mesenchymal stem cell ； Co-culture ； Side population

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