探討肺癌腦轉移之幾丁聚醣三維體外模型之建立與特性分析

Abstract

根據世界衛生組織(WHO)統計，肺癌近年來的發生率以及死亡率都是在全世界排名前三的癌症，而此癌症隨著時間的發展，大約會有三、四成的病患發現肺癌細胞有腦轉移的情形，這些轉移後的腫瘤細胞與原位癌細胞相比，其腫瘤的性質、特性會產生改變。 目前腦轉移細胞相關特性的研究仍以二維培養或動物實驗為主，然而，傳統的二維培養對於實際的生理環境機制可能沒辦法完全模擬，而減少動物實驗也是最近這幾年來有許多國家在推廣的計畫，因此，本研究期望可以建立一個能替代動物實驗、且能模擬腦轉移細胞生理環境的平台，並期望能透過此平台來分析其相關特性以及藥物篩選。 本研究利用細胞對於幾丁聚醣低貼附的特性，來建立三維細胞的培養環境，並將肺癌細胞A549與在腦轉移過程中相當重要的角色──星狀細胞CTX-TNA2進行共培養，並觀察在三維培養中共培養細胞的相關性質，如：細胞活性、蛋白表現等。而研究結果發現，我們在三維環境下所構成的共培養細胞球的抗藥性相對於傳統二維培養還要高，且在某些細胞蛋白的表現也有特定的成果。 整體而言，本研究成功地透過幾丁聚醣的特性建立了三維細胞體外模型，提供針對肺癌腦轉移一個新的特性分析、藥物治療的研究平台，未來也期望能將三維模型套用在不同類型的癌細胞腦轉移(如：乳癌)，為腦轉移相關癌症研究、治療做出貢獻。

According to the statistics from the World Health Organization (WHO), lung cancer has ranked among the top three cancers worldwide in terms of both incidence and mortality rates in recent years. With disease progression, approximately 30% to 40% of patients are found to have brain metastases of lung cancer cells. These metastatic cells undergo changes in their characteristics compared to the primary cancer cells. Currently, research on the characteristics of brain metastatic cells mainly relies on 2D cell culture or animal experiments. However, traditional 2D cell culture may not fully mimic the physiological environment, and reducing animal experiments has been a goal of many countries in recent years. Therefore, this study aims to establish a platform that can serve as an alternative to animal experiments and simulate the physiological environment of brain metastatic cells. The platform is expected to be used for related characteristic analysis and drug screening. In this study, we utilized the low-adhesive properties of cells to chitosan to establish a three-dimensional cell culture environment. Lung cancer cells A549 and astrocytes CTX-TNA2, which play a crucial role in brain metastasis, were co-cultured to observe the relevant characteristics of cells in a three-dimensional environment, such as cell viability and expression of protein. The experimental results showed that the drug resistance of co-culture spheroids formed in the three-dimensional environment was higher compared to traditional two-dimensional culture, and there was different expression of relevant protein. The results of this study successfully established a three-dimensional cell in vitro model based on chitosan, providing a new platform for characteristic analysis and drug treatment research for lung cancer brain metastasis. In the future, it is also expected to apply the three-dimensional model to other cancer brain metastases (such as breast cancer), contributing to research and treatment of brain metastasis-related cancers.