Differential expression of genes associated with cell proliferation and apoptosis induced by okadaic acid during the transformation process of BALB/c 3T3 cells
Differential expression of genes associated with cell proliferation and apoptosis induced by okadaic acid during the transformation process of BALB/c 3T3 cells
复制标题
DOI：
10.1016/j.tiv.2007.08.013
复制
发表时间：
2008-02-01
影响因子：
3.2
通讯作者：
Cao, Jia
中科院分区：
文献类型：
--
作者：
Ao, Lin;Liu, Jin-yi;Cao, Jia
冈田酸（OA）在两期癌变实验中是一种肿瘤启动子。然而，OA对细胞转化、细胞增殖和细胞凋亡的影响差异很大，而OA这些影响背后的分子事件尚不清楚。在本研究中，我们研究了OA在BALB/c 3T3细胞中促进细胞生长和凋亡的活性及其相关作用，并通过微阵列分析评估了基因转录表达的变化。OA的促进活性通过两阶段转化试验来评估，在两阶段转化试验中，细胞首先用低剂量的n -甲基-n '-硝基-n -亚硝基胍（MNNG）引发剂处理，然后用OA处理14天。结果表明，7.8 ~ 31.3 ng/ml浓度的OA能促进mnng处理的细胞转化。在促进期，暴露于OA （7.8 ng/ml）的细胞在前2天生长缓慢，随后死亡。Hoechst 33342荧光染料和Annexin-V/PI双色流式细胞术检测，OA诱导细胞形态凋亡，增加早期凋亡细胞比例。利用含有1796个克隆的小鼠毒理学芯片，测定了OA在促进期5个时间点诱导的基因表达谱，共鉴定出177个差异表达基因。通过基因本体论分析，其中31个被确定在功能上参与细胞生长和/或维持。在该组中，许多与细胞增殖和细胞周期进展相关的基因在早期和/或中期时间点下调。其中有一个与细胞凋亡相关的基因子集，其中Bnip3、Cycs、Casp3和Bag1基因参与细胞凋亡的线粒体途径。Ier3、Mdm2和Bnip3基因可能是p53的靶点。此外，real-lime PCR证实了从差异表达基因中随机选择的5个基因的表达变化，我们认为OA在两阶段，mnng启动的BALB/c 3T3细胞转化中诱导细胞生长抑制和凋亡。基因表达谱分析结果表明，oa诱导的细胞增殖抑制和细胞凋亡涉及多种分子途径。线粒体和p53相关的凋亡途径也可能有助于oa诱导的细胞凋亡。(C) 2007 Elsevier Ltd.版权所有。
Okadaic acid (OA) is a tumor promoter in two-stage carcinogenesis experiments. Nevertheless, the effects of OA on cell transformation, cell proliferation and apoptosis vary widely, and the molecular events underlying these effects of OA are not well understood. In the present study, we examined the promoting activity and the associated effects on cell growth and apoptosis mediated by OA in BALB/c 3T3 cells, and evaluated alterations of gene transcriptional expression by microarray analysis. The promoting activity of OA was estimated by a two-stage transformation assay, in which cells were treated first with a low dose of the initiator N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and then with OA for 14 days. It showed that OA, at concentrations of 7.8-31.3 ng/ml, enhanced the transformation of MNNG-treated cells. In the promotion phase, cells exposed to OA (7.8 ng/ml) grew slowly for the first 2 days and subsequently died. As determined by Hoechst 33342 fluorescent dye and Annexin-V/PI dual-colored flow cytometry, OA induced morphologically apoptotic cells and increased the percentage of early apoptotic cells. The gene expression profile induced by OA at five time points in the promotion phase was determined by use of a specific mouse toxicological microarray containing 1796 clones, and a total of 177 differentially expressed genes were identified. By gene ontology analysis, 31 of these were determined to be functionally involved with cell growth and/or maintenance. In this group, numerous genes associated with the cell proliferation and cell cycle progression were down-regulated at early and/or middle time points. Among these was a subset of genes associated with apoptosis, in which Bnip3, Cycs, Casp3 and Bag1 genes are involved in the mitochondrial pathway of apoptosis. Ier3, Mdm2 and Bnip3 genes may be p53 targets. Furthermore, real-lime PCR confirmed the expression changes of five genes selected at random from the differentially expressed genes, We conclude that OA induces cell growth inhibition and apoptosis in the two-stage, MNNG-initiated transformation of BALB/c 3T3 cells. The results of gene expression profile analysis imply that multiple molecular pathways are involved in OA-induced proliferation inhibition and apoptosis. Mitochondrial and p53-associated apoptotic pathways also may contribute to OA-induced apoptosis. (C) 2007 Elsevier Ltd. All rights reserved.