细胞色素P450花生四烯酸环氧化酶对肿瘤细胞增殖的影响。

Effects of cytochrome P450 arachidonic acid epoxygenases on the proliferation of tumor cells.

作者信息

Jiang Jian-Gang, Shen Gui-Fen, Chen Chen, Fu Xian-Ning, Wang Dao-Wen

机构信息

Department of Internal Medicine, Tongji Hospital, Tongji Medical School, Huazhong University of Science and Technology, Wuhan, Hubei, China.

出版信息

Ai Zheng. 2009 Jan;28(1):14-9. Epub 2009 Jan 5.

PMID:19448409

Abstract

BACKGROUND AND OBJECTIVE

Cytochrome P450 (CYP) arachidonic acid epoxygenases promote cell proliferation and inhibit apoptosis in endothelial cells. This study was to investigate the effects of CYP epoxygenases 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 epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase (ERK)1/2 and Akt before and after transfection were detected by western blot. Tca-8113 cells infected with rAAV-CYP2J2, rAAV-CYPF87V, rAAV-antiCYP2J2 and rAAV-GFP were inoculated into nude mice, to observe the effect of CYP epoxygenases on the growth of xenografts.

RESULTS

Infection of Tca-8113, A549, Ncl-H446 and HepG2 cells with rAAV-CYP2J2 and rAAVCYPF87V significantly increased the proliferation of tumor cells by 1.7-, 1.4-, 1.6- and 2.2-fold, and 2.0-, 1.5-, 1.8- and 2.0-fold respectively, as compared with control cells. On the contrary, infection with rAAV-antiCYP2J2 inhibited the proliferation of the four tumor cell lines. Moreover, CYP epoxgenases remarkably enhanced phosphorylation of EGFR, ERK1/2 and Akt, and upregulated total PI3K by 2-, 2.3-, 2.4- and 1.9-fold in the four cell lines, while rAAV-antiCYP2J2 exerted an inhibition effect. Infection of CYP450 epoxygenase genes markedly increased the cell percentage in S/G(2)/M phases by 210% as compared to control Tca-8113 cells. rAAV-CYP2J2 and rAAV-CYPF87V promoted tumor growth of Tca-8113 cell xenografts in nude mice in comparison to the control and rAAV-antiCYP2J2 groups.

CONCLUSION

CYP epoxygenases efficiently promote the proliferation of tumor cells, which may be related with the activation of EGFR, ERK1/2 and PI3K/Akt signaling pathways.

摘要

背景与目的

细胞色素P450（CYP）花生四烯酸环氧化酶可促进内皮细胞增殖并抑制其凋亡。本研究旨在探讨CYP环氧化酶对肿瘤细胞增殖的影响及其可能的信号通路。

方法

采用MTT法和流式细胞术检测重组腺相关病毒（rAAV）介导的细胞色素P450 2J2（CYP2J2）、细胞色素P450 F87V（CYPF87V）及抗CYP2J2对Tca-8113、A549、Ncl-H446和HepG2细胞增殖的影响。通过蛋白质印迹法检测转染前后磷酸化表皮生长因子受体（EGFR）、细胞外信号调节激酶（ERK）1/2和Akt的表达。将感染rAAV-CYP2J2、rAAV-CYPF87V、rAAV-antiCYP2J2和rAAV-GFP的Tca-8113细胞接种于裸鼠，观察CYP环氧化酶对异种移植瘤生长的影响。

结果

与对照细胞相比，rAAV-CYP2J2和rAAV-CYPF87V感染Tca-8113、A549、Ncl-H446和HepG2细胞后，肿瘤细胞增殖显著增加，分别为1.7倍、1.4倍、1.6倍和2.2倍，以及2.0倍、1.5倍、1.8倍和2.0倍。相反，rAAV-antiCYP2J2感染抑制了这四种肿瘤细胞系的增殖。此外，CYP环氧化酶显著增强了四种细胞系中EGFR、ERK1/2和Akt的磷酸化，并使总PI3K上调了2倍、2.3倍、2.4倍和1.9倍，而rAAV-antiCYP2J2则发挥抑制作用。与对照Tca-8113细胞相比，CYP450环氧化酶基因感染使S/G(2)/M期细胞百分比显著增加了210%。与对照组和rAAV-antiCYP2J2组相比，rAAV-CYP2J2和rAAV-CYPF87V促进了裸鼠体内Tca-8113细胞异种移植瘤的生长。