佛波酯对BALB/c3T3细胞转化早期基因表达的影响  

Gene expression profiles in early stage of B ALB/c 3T3 cells’transformation promoted with 12-O-tetradecanoylphorbol-13-a cetate

作　　者：敖琳[1] 曹佳[1] 黄明辉[2] 伍亚舟[3] 刘晋祎[1] 张彦琦[3] 曾志雄[2] 杨梦苏[2] 

机构地区：[1]第三军医大学预防医学系卫生毒理学教研室,重庆400038 [2]香港城市大学基因组科技应用研究中心 [3]第三军医大学预防医学系卫生统计学教研室,重庆400038

出　　处：《中华预防医学杂志》2005年第2期99-102,F003,共5页Chinese Journal of Preventive Medicine

基　　金：国家973计划资助项目(2002CB512901);国家自然科学基金资助项目(30271136;30200354);国家杰出青年基金资助项目(30125037)

摘　　要：　目的　探讨佛波酯(12- O -tetradecanoylphorbol- 13 -acetate,TPA)促进细胞转化早期基因表达谱的变化。方法　以N 甲基 N’ 硝基 N 亚硝基胍 (N methyl N’ nitro N nitrosoguanidine,MNNG)为启动剂,TPA为促癌剂,建立BALB/c3T3细胞转化模型。采用锥虫蓝染色法检测细胞生长情况,流式细胞仪检测细胞周期变化。采用cDNA微阵列检测TPA处理早期的基因表达谱变化。结果　TPA处理早期可抑制细胞增殖,阻滞细胞于G1 期与S期。TPA处理 4h和 24h后,在检测的1 152个基因中筛选出 19个差异表达基因,其中 9个基因表达上调, 10个基因表达下调。许多差异表达基因的功能与细胞增殖、凋亡和周期调控相关,主要涉及ras和p53基因信号传导通路。结论　TPA在促进BALB/c3T3细胞转化的早期阶段,可影响某些调节细胞周期进展基因的转录表达,从而导致细胞生长阻滞。Objective To elucidate the po tential molecular mechanism responsible for the early time of tumor promotion, g ene expression profile was studied in the transformed BALB/c 3T3 cells induced b y 12-O-tetradecanoylphorbol-13-acetate(TPA).Methods The two-stage cell transformation model was established by using the initiato r of N-methyl-N’ -nitro-N-nitrosoguanidine (MNNG) and promoter of TPA. C ell proliferation was measured by trypanblau staining and cell cycle analysis wa s carried out by flow cytometry assay.A cDNA microarray representing 1 152 gen es was used to investigate the gene expression profiles of BALB/c 3T3 cells expo sed to TPA at 4 h and 24 h respectively.Results TPA coul d effectively inhibit cell proliferation and induce the G 1 and S cell cycle ar rested in the early time.Moreover 19 genes were found differentially expressed at least twofold in the TPA treated cells as compared with the control cells, 9 of them were upregulated and 10 downregulated.Most of the differentially expr essed genes were involved in cell proliferation, differentiation or apoptosis, a nd related to ras or p53 signal transduction pathway.Conclusion TPA could influence the transcriptional expression of some genes related to cell cycle modulation and ultimately result in the cell growth arrest.

关 键 词：TPA 早期 BALB/C 基因表达谱 佛波酯 3T3细胞 细胞增殖 差异表达基因 细胞生长 转化 

分 类 号：R73-3[医药卫生—肿瘤]