輻射照射腦瘤細胞所致之旁觀者效應以及肺癌相關重要基因之功能性探討

Radiation-Induced Bystander Effect in Human Glioblastoma Cells and Functional Characterization of Lung Cancer Related Genes

侯雅雪(Ya-Hsiue Hou)

指導教授 ： 江士昇 ； 楊素珍

中臺科技大學/健康科學院/放射科學研究所/碩士(2008年)

摘要

肺癌為全球常見之癌症之一，為國內癌症死因之首，其中以非小細胞肺癌(NSCLC)居多，形成癌症之因素眾多複雜，目前沒有良好的篩檢工具、有效的治療方法，是造成肺癌存活率、治癒率低的主因。獲得肺癌的早期診斷或存活之生物標幟，應可開發有效肺癌的篩檢方法，或在臨床治療上提高病人存活率。為獲得這些生物標幟，我們以微陣列基因晶片在肺癌細胞株篩選出sox9基因。sox9為SOX家族轉錄因子在生物發育之過程及癌症中扮演重要角色，為探討sox9在NSCLC中的功能性，在本篇論文我們以data mining及siRNA干擾技術剔除sox9基因再以分子生物技術進行分析。實驗結果發現，sox9在臨床NSCLC組織中之表現較鄰近或正常組織高；在NSCLC復發病人之表現量較沒有癌復發者高。剔除sox9會降低NSCLC細胞株之cell viability；降低NSCLC細胞株之G1 phase、增加G2 phase；降低肺腺癌細胞株之存活率；增加肺腺癌細胞株之侵犯力。以微陣列基因晶片進行分析sox9剔除與哪些訊息傳遞路徑有關，用Quantitative real-time RT- PCR(即時定量反轉錄聚合鏈鎖反應)加以驗證，結果為MT2A 、FTL 、CASP4 、FTH1 、RPL35 、AKAP5 、JNK1 皆被過度表現；sox1 、sox17 、BBC3 、IRS2 、WNT6 皆為低表現這些基因之功能性主要與細胞訊息之傳遞及細胞生長有關。我們認為sox9 與調控NSCLC 之細胞存活是有重要關聯的。

關鍵字

微陣列基因晶片 ； 存活率 ； 肺癌 ； sox9

並列摘要

Lung cancer, especially non-small cell lung cancer (NSCLC), is the most common cause of cancer related death worldwide. It is generally believed that lack of biomarkers of early detection or prognosis of NSCLC is the main reason that explains high mortality of this disease. In order to obtain such biomarkers, we have conducted a lung cancer cell line-based functional genomics screen, in which sox9 was a candidate biomarker. In the present study, in silico data mining and molecular biology emphasizing siRNA gene silencing technique was used to probe the functional roles of sox9 in NSCLC. Here we showed that the expression of sox9 is significantly higher in the tumor parts than in the normal parts of cells of NSCLC; mean while, its expression is also higher in patients with recurrent diseases than those without. Furthermore, silencing of sox9 expression can cause reduced viability of cells, affect the cell cycle progression through G1 or G2 phases, and change the invasion capability of cells. Microarray gene expression analysis and quantitative polymerase chain reaction (q-PCR) revealed that the genes affected by silencing of sox9 included MT2A、FTL、CASP4、FTH1、RPL35、AKAP5、JNK1 (up-regulated) and sox1、sox17、BBC3、IRS2、WNT6 (down-regulated). Many of the differentially expressed genes are involved in the signaling pathways of cell growth or proliferation. In summary, we concluded that sox9 plays important roles in the survival of NSCLC cells.

並列關鍵字

cell survival ； microarray analysis ； lung cancer

參考文獻

1 行政院衛生署網站

3 Chu YW, Yang PC, Yang SC, Shyu YC, Hendrix M J. C., Wu R, and Wu CW. Selection of Invasive and Metastatic Subpopulations from a

Human Lung Adenocarcinoma Cell Line. Am. J. Respir. Cell Mol. Biol. Vol. 17, pp. 353–360, 1997

5 Bi W, Deng JM, Zhang Z, Behringer RR, de Crombrugghe B. Sox9 is required for cartilage formation. Nat Genet. 1999 May;22(1):85-9.

6 Mori-Akiyama Y, Akiyama H, Rowitch DH, de Crombrugghe B. Sox9 is required for determination of the chondrogenic cell lineage in the cranial neural crest. Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9360-5. Epub 2003 Jul 23.