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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 investigate the changes of early gene expression profiles induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). Methods BALB / c3T3 cell transformation model was established by using N methyl N ’nitro N nitrosoguanidine (MNNG) as a promoter and TPA as a cancer promoting agent. Cell growth was detected by trypan blue staining, and cell cycle was detected by flow cytometry. The cDNA microarray was used to detect the changes of gene expression profile in the early stage of TPA treatment. Results TPA inhibited cell proliferation in early stage and arrested cells in G1 phase and S phase. After 4 and 24 h of TPA treatment, 19 differentially expressed genes were screened out among the 1 152 genes tested, among which 9 genes were up-regulated and 10 genes were down-regulated. The function of many differentially expressed genes is related to cell proliferation, apoptosis and cycle regulation, mainly involving the ras and p53 gene signaling pathways. Conclusion TPA can promote the transcriptional expression of certain genes that regulate the cell cycle progression in the early stages of BALB / c3T3 cell transformation, leading to cell growth retardation.