中文摘要：

背景与目的：细胞色素P450（cytochromeP450，CYP）花生四烯酸表氧化酶对内皮细胞具有促进增殖、抑制凋亡的作用。本研究主要探讨CYP表氧化酶对肿瘤细胞增殖的影响，并初步探讨其影响细胞增殖的相关信号转导机制。方法：用重组腺相关病毒（recombinant adeno-associatedvirus，rAAV）介导CYP花生四烯酸表氧化酶基因CYP2J2（cytochromeP4502J2）、CYPF87V（cytochromeP450F87V）和反义CYP2J2基因分别转染A549、Tea-8113、HepG2、Ncl-H4464种肿瘤细胞．采用MTT法、细胞计数法和流式细胞术分析CYP表氧化酶对肿瘤细胞增殖的影响：并用Westernblot法检测转染前后Tca-8113细胞中表皮生长因子受体（epidermal growth factor receptor，EGFR）、ERKI／2和Akt磷酸化水平：最后将转染rAAV-CYP2J2、rAAV-CYPF87V、rAAV-antiCYP2J2和rAAV-GFP的Tca-8113细胞分别接种至裸鼠皮下，观察皮下移植瘤的牛长情况。结果：转染CYP2J2和CYPF87V后，A549、Tca-8113、HepG2和Ncl-H446细胞的细胞增殖数分别是相应未转染细胞的1．7倍和2．0倍、1．4倍和1．5倍、1．6倍和1．8倍、2．2倍和2．0倍；转染反义CYP2J2阻断细胞自身CYP2J2表达后，上述4种细胞增殖显著减低：流式细胞术结果显示转染CYP表氧化酶基fj；I的肿瘤细胞中S／G2／M期细胞增加了210％。Westernblot结果显示，转染CYP2J2基因后，EGFR、ERK1／2和Akt磷酸化水平分别为未转染细胞的2倍、2．3倍和2．4倍，同时P13K表达水平也L调为未转染细胞的1．9倍：相反．转入反义CYP2J2基因明显抑制这些蛋白的磷酸化和表达。在体皮下移植瘤实验结果显示，转染rAAV-CYP2J2、rAAV—CYPF87V组裸鼠皮下移植瘤出现的时间（分别为5．8d和6．0d）明显较对照组和转染rAAV—GFP组（分别为8．4d和8．6d）短：而转染反义rAAV-CYP2J2组移植瘤生长缓慢，出现时间也明显较长（约10d）。结论：CYP花生四烯酸表氧化酶能促进肿瘤细胞

英文摘要：

Background and Objective= Cytochrome P450 （CYP） arachidonic acid epoxygenase promotes cell proliferation and inhibits apoptosis in endothelial cells. This study was to investigate the effects of CYP epoxygenase on the proliferation of tumor cells and possible signaling pathways. Methods： The effects of recombinant adeno-associated virus （rAAV） mediated cytochrome P450 2J2 （CYP2J2）, cytochrome P450 F87V （CYPF87V） and anti-CYP2J2 on proliferation of Tca-8113, A549, Ncl-H446 and HepG2 cells were measured using MTT and flow cytometry. Expressions of phosphorylated EGFR, ERK and Akt after the treatment with CYP2J2 were detected by Western blot. Tca-8113 cells infected with rAAV-CYP2J2, rAAV- CYPF87V, rAAV-anti-CYP2J2 and rAAV-GFP were inoculated into nude mice, to observe the effect of CYP epoxygenase on the growth of xenografts in nude mice. Results; Infection of Tca-8113, A549, NcI-H446 and HepG2 cells with rAAV-CYP2J2 and rAAV-CYPF87V significantly increased the proliferation of tumor cells to 1.7, 1.4, 1.6 and 2.2 folds, as compared with control cells. On the contrary, infection with rAAV-anti-CYP2J2 inhibited the proliferation of the four tumor cell lines. Moreover, CYP epoxgenase remarkably enhanced phosphorylation of EGFR, ERK1/2 and Akt, and upregulated the expression of PI3K to 2, 2.3, 2.4 and 1.9 folds in the four cell lines, while rAAV-anti-CYP2J2 exerted an inhibition effect. Infection of CYP450 epoxygenase genes markedly increased the cell percentage in S/ GJM phase as compared to control Tca-8113 cells （63.77% vs. 29.90%）. rAAV-CYP2J2 and rAAV-CYPF87V promoted tumor growth of Tca-8113 cell xenografts in nude mice in comparison to the control groups. Conclusion： CYP epoxygenase could efficiently promote the proliferation of tumor cells, which may be related with the activation of EGFR, ERK1/2 and PI3K/Akt signaling pathways.