- 學位論文
以微晶片實驗探討天生靈芝乙醇萃液對非小細胞肺癌細胞株的抗癌作用
A Gene Chip Approach to Anti-cancer Effect of Tien-Shen Lingzhi Ethanol Extracts on Non-Small Cell Lung Cancer Cell Line
摘要
背景：肺癌是所有惡性腫瘤疾病中預後最差的疾病之一，仍必須尋求更多的治療方法以改善臨床預後。從傳統中草藥尋找具有生物活性之治療成分已經成為一種新趨勢。幾個世紀以來，靈芝一直是廣被使用作為促進健康的中草藥。我們曾經發表過關於靈芝在不同類型癌症的抗癌功效，本研究以基因體實驗進一步探討天生靈芝(台灣松杉靈芝改良品種)抗癌作用相關的訊息傳導路徑。方法：以Affymetrix Human Genome U133 Plus 2.0 晶片設計實驗，藉由生物資訊工具GeneGo Meta CoreTM 分析比較，尋找非小細胞肺癌細胞株H23/0.3 經天生靈芝處理與否所引起的差異性表達基因。結果：非小細胞肺癌細胞株H23/0.3接受天生靈芝處理與否的全基因體表達， 由多元尺度分析(multidimentional scaling)呈現明顯之差異。生物資訊分析顯示，天生靈芝擾亂了非小細胞肺癌細胞株的細胞週期，且細胞週期調控相關的訊息傳導路徑是最主要被影響之標的。在26 個與DNA 合成相關的訊息傳導分子中，有17 個在本次分析中呈現差異。針對部分差異表達基因以Q-PCR 實驗驗證，結果亦與晶片實驗結果一致。再以流式細胞儀檢測，顯然地，經天生靈芝處理之肺癌細胞株呈現細胞週期停滯。結論：微晶片實驗顯示，天生靈芝擾亂非小細胞肺癌細胞株H23/0.3 的細胞週期，流式細胞儀檢測到的細胞週期擾亂與晶片生物資訊分析結果是一致的。
並列摘要
Background: Lung cancers are among the neoplastic diseases with the worst prognosis. More treatment modalities are needed to improve clinical outcome. It’s being a new trend to search for bioactive traditional Chinese medicines (TCM). Ganoderma, also known as Lingzhi, is a TCM and has been widely used for improving human health for centuries. We have addressed the anticancer effects of Ganoderma on various cancers. In this study, a comprehensive genomic profiling was used to further explore the signal pathway for the anticancer effects of Tien Shen Lingzhi(TSL),an improved strain of Ganoderma tsugae. Methods: A comparative genomics study was performed to search for TSL-mediated differential expressions on H23/0.3 cells. Bioinformatics tool, e.g., GeneGo Meta CoreTM ,was used to analyze the experimental results based on Affymetrix Human Genome U133 Plus 2.0 microarray. Results: The whole-genome expression profiles of H23/0.3 with or without TSL treatment were resolved by multidimentional scaling (MDS) analysis. It indicated a TSL-mediated cell cycle perturbation. Bioinformatics analysis showed cell cycle was the most affected pathway. GeneGo Meta CoreTM derived cellular pathway map revealed dramatic hits (17/26) on the signal transduction of DNA replication. The microarray results were confirmed by Q-PCR. Conclusion: The genomic microarray data demonstrates the TSL-induced cell cycle deregulation in H23/0.3 cells. TSL-mediated cell cycle perturbation on H23/0.3 cells is consistent with the cellular pathway map resulting from GeneGo Meta CoreTM analysis.