学术沙龙汇报.pptxVIP

CREB1 directly activates the transcription of ribonucleotide reductase small subunit M2 and promotes the aggressiveness of human colorectal cancer Oncotarget Advance Publications 2016 (RR) CREB1 1、cAMP responsive element binding protein 1 （环磷腺苷效应元件结合蛋白1）； 2、调节基因转录，转录增强因子； 3、CREB1异常表达(白血病、乳腺癌、非小细胞肺癌、肾癌)； 4、参与细胞增殖、迁移和侵袭； 5、调节肿瘤相关基因（Bcl-2,c-fos,TNF-α）。 1、Ribonucleotide reductase； 2、催化核糖核苷酸为脱氧核糖核酸； 3、由RRM1和RRM2/RRM2B组成； 4、RRM1-RRM2； 5、RRM1-RRM2B。 CREB1 increases RRM2 expression in CRC cells Figure 1： A. HCT116, HT29, and RKO cells were transfected with either control siRNA or CREB1 siRNA for 48 h, and harvested for Western blot analysis with antibodies anti-CREB1, anti- RRM2, and anti- GAPDH (as loading control). B. HCT116, HT29, and RKO cells were transfected with control or CREB1 siRNA for 48 h. The mRNA levels were analyzed by qPCR (normalized by actin). *P 0.05 1 CREB1 increases RRM2 expression in CRC cells Figure 1： C. HCT116 cells were seeded onto the coverslips in culture dishes. Cells were transfected with indicated siRNA or expression plasmids for 48 h, fixed and then immunoflourescence assay was performed. DAPI served as nuclear marker. 1 CREB1 increases RRM2 expression in CRC cells Figure 1： D. HCT116 or RKO cells were transfected with empty vector (EV) or CREB1 expression plasmid, and then harvested for Western blots and RNA expression analysis 48 h later. *P 0.05. E. HCT116 or RKO cells were transfected with control siRNA or CREB1 siRNA as well as RRM2 reporter (-2465/+23) and an internal control reporter pRL-TK for 48 h. *P 0.05. 1 CREB1 directly binds to RRM2 promoter and induces its transcription Figure 2： A. Relative luciferase activity in HCT116 cells co-transfected with different amounts of E2F1 expression plasmid, RRM2 promoter reporter (-2465/+23), and an internal control reporter pRL-TK. B. The graph shows three putative cyclic-AMP response elements (CREs) on the RRM2 promoter, where CREB1 could potentially bind to and activate RRM2 tran