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標題: 懸浮微粒脂溶性萃取物誘發人類乳癌細胞及肺纖維母細胞之毒性和DNA損害
Induction of cytotoxicity and DNA damage in human breast carcinoma cells and lung embryonal cells by the organic extracts of particulate matter ( PM )
作者: 林家琦
Lin, Chia-Chi
關鍵字: Particulate matters;懸浮微粒;cytotoxic;ROS;DNA damage;breast cancer cells;細胞毒性;氧化壓力;DNA損壞;乳癌細胞
出版社: 環境工程學系所
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Lett. , 122, 103-110. 莊秉潔(2006),台灣火力電廠環境空氣品質平行監測期末報告 莊惠萍(2005),屏東地區空氣污染對易感性族群健康影響之研究,高雄醫學大學公共衛生學研究所碩士論文。 郭育良等人(2005),職業病概論,華杏出版股份有限公司。 黃俊智(2000),焚化爐衍生空氣污染物特性分析之研究,國立屏東科技大學環境工程與科學系碩士學位論文。 黃群真(2006),戴奧辛誘發人類乳癌細胞株(MCF-7及MDA-MB-231 cells)細胞毒性及核酸氧化損壞作用研究,國立中興大學環境工程學研究所碩士論文。 蔡尚學(2003),台灣地區室外空氣污染對健康效應之研究,高雄醫學大學醫學研究所博士論文。 謝振友(1999),台北市垃圾焚化爐排氣對周界懸浮微粒之影響評估,國立臺灣大學環境工程學研究所碩士論文。
懸浮微粒(particulate matter, PM)已知能對人類誘發廣泛之生物毒性效應。累積之研究證據顯示,PM中之內含物質,如多環芳香族碳氫化物(polyaromatic hydrocarbons, PAHs) 及過渡金屬,誘發之氧化壓力(oxidative stress)及核酸損壞作用於PM所誘發之致細胞毒性及致癌機制上扮演一個重要的角色。由於氧化壓力是造成老化、自發性突變、及癌症生成之主要因素,因此探討不同來源之PM誘發氧化壓力之機制及其對細胞毒性作用之影響,將有助於了解PM致毒及致癌作用機轉。本研究計畫主要目標是探討曝露於PM之脂溶性萃取物環境下,能否經由代謝活化及誘發基因表現失衡,進而導致細胞氧化壓力、核酸氧化損壞作用及細胞死亡。本研究以人類細胞運用生物性指標,包括細胞存活率測試(SRB assay)、氧化壓力測定法(oxidative burst assay)、DNA斷鏈分析方法(Comet assay)、核酸醛基損害分析法(ADL assayed by ARP-ASB)和及時定量(real-time RT-PCR)分析法,以了解懸浮微粒所造成之細胞毒性、氧化壓力、DNA損害、及其所誘發之基因表現。由研究結果顯示,各採樣站及各時間點之總懸浮微粒(TSP)於四株細胞MDA-MB-231、MCF-7、MRC-5、與XPA細胞所誘發之細胞毒性皆具有時間及劑量之效應。TSP於24h即可使MDA-MB-231和MCF-7細胞內累積1.06~9.67倍之ROS,此外,ERα會影響由TSP脂溶性萃取液對細胞誘發之毒性相關效應。在TSP誘發細胞中DNA損害方面,皆可使MDA-MB-231和MCF-7細胞誘發DNA單股斷鏈。MCF-7人類乳癌細胞於東大採樣站(2005/12/05)和草屯採樣站(2005/12/05)反應處理後CYP 1A1 mRNA基因表現有明顯增加作用(p < 0.05);CYP 1B1 mRNA則皆無明顯基因表現增加作用。因此綜合本研究結果顯示,台灣中部地區之懸浮微粒會誘發人類乳癌細胞MCF-7之CYP 1A1 mRNA基因表現增加,造成ROS之生成及累積,進而誘發細胞毒性及DNA損壞。

Particulate matters (PM) are known to induce a broad-spectrum of adverse biological effects in humans. Cumulating evidence indicates that solvent-extractable organic compounds from PM, including polycyclic aromatic hydrocarbons (PAHs) and transition metals play an important role in PM-induced oxidative stress and the subsequent induction of DNA lesions in mammalian cells. As oxidative stress is the source of aging, endogenous mutation, and cancer, investigation in the induction of oxidative stress and DNA lesions by PM should shed light on the mechanism(s) by which PM exert their actions. The primary goal of this study is to examine whether exposure to the organic extracts of PM may induce oxidative stress, DNA lesions, and cell death via the induction of imbalance in gene expression. To achieve these research goals, we applied biochemical assays to assess the extent of cell death, DNA lesions, and altered gene expression induced by the organic extracts of PM in human cultured cells. Results from this research should provide pivotal information of the relevance of the environmental exposure to PM and the mechanism(s) by which PM induce cytotoxic response and oxidative damage to genomic DNA. Results from cytotoxicity analyses indicated that the organic extracts of total suspended particles(TSP) induced concentration- and time-dependent increases in cytotoxic response in MDA-MB-231, MCF-7, MRC-5 and XPA cells. Further, we observed that TSP induce ~1.06-9.67-fold increases in ROS formation in MDA-MB-231 and MCF-7 cells after 24h of exposure. Overall, these findings suggest that the status of estrogen receptor may play a role in modulating the induction of oxidative DNA damage and cell death by organic extracts of TSP in human breast cancer cells. The data also reveal that the organic extracts of TSP was capable of inducing DNA single strain break in both MDA-MB-231 and MCF-7 cells as measured by the single-gel-electrophoresisi (Comet) assay. Further investigation indicated that TSP(from Tunghai university and Caotun)induced increases in the expression of CYP1A1 genes in MCF-7 cells as determined by the real-time RT-polymerase chain reaction (real-time RT- PCR) assay. Overall, results from our current study strongly suggests that the organic extracts of PM collected at three sampling sites in central Taiwan are capable of inducing cytotoxic response and DNA damage in human MCF-7 cells through altered gene expression and the subsequent induction of oxidative stress.
其他識別: U0005-1408200712325500
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