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目的用基因芯片观察花色苷矢车菊素-3-葡萄苷(C3G)作用人白血病单核细胞细胞株(THP-1)源性巨噬细胞前后相关信号通路上基因表达的差异。方法100μmol/L的C3G作用于THP-1源性巨噬细胞12h,用信号转导通路发现者基因芯片研究C3G对THP-1源性巨噬细胞基因表达谱的影响,并利用半定量逆转录-聚合酶链反应(RT-PCR)法对筛选出的部分基因进行验证。结果C3G作用后有7条信号通路中的15个基因发生改变,其中表达上调的基因有5个,包括肿瘤基因p53(TP53)、核因子κB抑制因子α(NFκBIA)、视黄醇结合蛋白基因2(RBP2)等;表达下调的基因有10个,包括FBJ鼠科骨肉瘤病毒癌基因同源物(V-fos)、诱导型一氧化氮合酶(NOS2A)、前列腺素内过氧化物合酶2(PTGS2)等,这些基因编码的宿主蛋白均为肿瘤免疫、炎症反应相关的信号转导通路中的关键点。半定量RT-PCR结果显示,经C3G干预后,NOS2A和PTGS2的表达水平下调了58%和54%。结论花色苷类植物化合物可能通过促有丝分裂、果蝇无翼基因同源物(WNT)、p53、磷脂酶C、核因子κB(NFκB)、Janus激酶/信号转导和转录活化因子(JAK/STAT)和维甲酸等信号转导通路发挥其抗炎、抗肿瘤功能。
Objective To observe the difference of gene expression in the signal pathways of human leukemia cell line THP-1-derived macrophages before and after using anthocyanin cyanidin-3-glucoside (C3G) by gene chip. METHODS: The THP-1-derived macrophages were treated with 100μmol / L of C3G for 12h, and the effects of C3G on THP-1-derived macrophages were investigated by using gene chip of signal transduction pathway. The selected genes were verified by polymerase chain reaction (RT-PCR). RESULTS: Fifteen of the seven signaling pathways changed after C3G treatment. Five of the seven genes were upregulated, including p53 (TP53), NFκBIA (α), retinol binding protein 2 (RBP2), etc. There were 10 genes that were downregulated, including FBJ murine osteosarcoma oncogene homolog (V-fos), inducible nitric oxide synthase (NOS2A), prostaglandin endoperoxide Enzyme 2 (PTGS2), etc. The host proteins encoded by these genes are the key points in the signal transduction pathways related to tumor immunity and inflammatory response. Semi-quantitative RT-PCR results showed that the expression levels of NOS2A and PTGS2 were down-regulated by 58% and 54% respectively after C3G intervention. Conclusions Anthocyanin compounds may be involved in mitosis, Drosophila wingless gene homolog (WNT), p53, phospholipase C, NFκB, Janus kinase / signal transducers and activators of transcription (JAK / STAT ) And retinoic acid and other signal transduction pathways to play its anti-inflammatory, anti-tumor function.