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研究生:李宜珍
研究生(外文):Yi Cheng Li
論文名稱:檳榔子萃取物對口腔細胞在型態與基因表現影響之分析
論文名稱(外文):Phenotype Characterization and Transcriptome Profiling of Oral Mucosa Cells Chronically Exposed to Areca Nut Extract
指導教授:鄭恩加
指導教授(外文):A. J. Cheng
學位類別:碩士
校院名稱:長庚大學
系所名稱:醫學生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
論文頁數:67
中文關鍵詞:口腔癌檳榔子
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口腔癌為台灣癌症死因排名的第六位,而其之高發生率與死亡率已知與嚼食檳榔子習慣息息相關。為了探討嚼食檳榔子誘發的致癌機轉,本研究建立長期低劑量曝露檳榔子萃取物的細胞株,藉此觀察其對細胞表型的變化以及參與分子層面影響。結果顯示,相較於母代細胞,經檳榔子處理的子代細胞生長能力降低了0.3-0.8倍,但細胞侵犯能力卻增強5-16倍;儘管不同細胞株的表現程度不甚一致,總體而言,經低劑量長期處理檳榔子的子代細胞對化療藥物與放射線照射有較高的抗性。進一步以cDNA微矩陣和生物資訊法演算分析母代細胞與子代細胞轉錄圖譜的差異,在p-value< 0.05下篩出相關的分子傳遞路徑,包含細胞恆定、免疫反應、細胞附著與壓力所導致訊息傳遞途徑。其中CHES1 mRNA在所有子代細胞株表現量皆下降,而過度表現CHES1則可能經由活化Erk導致細胞週期受抑制。臨床檢體分析顯示,在嚼食檳榔的口腔癌病患中,其腫瘤組織CHES1表現量相對於正常組織下降了0.19倍。以上研究直接提供檳榔子萃取物造成細胞表型及轉錄圖譜變化的證據,將有助於我們對口腔癌危險因子有更深入的認識。
Oral cancer is the 6th most frequent cancer in Taiwan. The habit of areca nut chewing is the main etiological factor of oral cancer. To shed light on the phenotype characterization and molecular basis of areca nut associated oral carcinogenesis, we established three oral cancer sublines chronically treated with areca nut extract (ANE) at IC70 dose for 2 months. Cell growth of the ANE subline was decreased, with 0.8 to 0.3 fold of reduction compared to the parental cells. Nevertheless, cell invasion ability was significantly increased in all three sublines, with approximately 5 to 16 folds of elevation. Although various levels in different cell lines, these ANE-sublines exhibited more resistance to stress (cisplatin and radiation) in general, that was associated with higher ROS clearance ability in the subline cells. cDNA microarrays were used in transcriptome profiling between parental and ANE sublines of oral cancer cells. Heretical cluster and algorithmic analyses of the microarray data revealed several pathways significantly associated with ANE (p< 10E-5), as homeostasis, immune response, cell adhesion and signaling pathway response to stimulus. Specially, CHES1 mRNA expressions were decreased in ANE sublines by RT-PCR assay. In addition, CHES1 overexpression arrested cell cycle at G2/M phase might through activating Erk pathway. Clinical study further showed that CHES1 was under expressed in oral cancer tissues. Therefore, the phenotype characterization and transcriptome profiling of oral mucosa cells chronically exposed to areca nut extract were determined. This study may contribute to clinical investigation as risk assessment.
BACKGROUND………………………………………………………...1
ORAL CANCER/ ORAL CAVITY CANCER……………………………….….1
RISK FACTOR OF ORAL CANCER………………………………………….2
BETEL QUID……………………………………………………………..3
ARECA NUT EXTRACT…………………………………………………...4
MULTIPLE CELLULAR EFFECTS IN RESPONSE TO ARECA NUT…………….7
CHES1…………..……………………………………………………...8
Study Aim and approach…………..…………………………………..10
MATERIALS AND METHODS…………..…………………………..11
RESULTS……………………………………………………………….20
DISSCUSION…………………………………………………………..28
REFERENCES…………………………………………………………32
TABLE AND FIGURE………………………………………………...41
1. Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncol 2009; 45: 309-16.
2. Chen YJ, Chang JT, Liao CT, Wang HM, Yen TC, Chiu CC, Lu YC, Li HF, Cheng AJ. Head and neck cancer in the betel quid chewing area: recent advances in molecular carcinogenesis. Cancer Sci 2008; 99: 1507-14.
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