Russo Ilaria, Carrizzo Albino, Bochicchio Sabrina, Piazza Ornella, Lamberti Gaetano, Barba Anna Angela, Vecchione Carmine, Zeppa Pio, Iovino Paola, Bucci Cristina, Santonicola Antonella, Ciacci Carolina
Department of Medicine and Surgery, Scuola Medica Salernitana, University of Salerno, 84081, Baronissi (SA), Italy, IRCCS Neuromed, Pozzilli (IS), 86077, Italy.
Department of Industrial Engineering, University of Salerno, via Giovanni Paolo II, 132, Fisciano (SA) - ITALY.
Transl Med UniSa. 2018 Mar 31;17:22-30. eCollection 2017 Jul.
Evidence in inflammatory bowel diseases (IBD) supports a connection between inflammation and cancer due to the alteration of the cell cycle with loss of control at the G1/S checkpoint. In this study, we analyze the expression and modulation of CyD1 and E2F1 in colon explants from Crohn's disease (CD) patients. We used ex vivo culture of colon explants from 4 CD patients and 2 healthy controls, stimulated with lipopolysaccharide from Escherichia Coli (EC-LPS). Commercial siRNAs for CyD1 and E2F1 inhibition were encapsulated in Invivofectamine® and in purposely produced nanoliposomal vectors to silencing CyD1 and E2F1 expression. Western blot analysis was used to investigate the effect of siRNA on CyD1, E2F1 and cyclooxygenase 2 (COX-2) expression. In CD patients colon explants, CyD1 and E2F1 increased after the inflammatory stimulus but siRNA silencing attenuated their expression and controlled the COX-2 expression too. These data represent a prelimiary exploration of in vitro siRNA use.
炎症性肠病(IBD)的证据支持炎症与癌症之间存在关联,这是由于细胞周期改变,在G1/S检查点失去控制所致。在本研究中,我们分析了克罗恩病(CD)患者结肠外植体中细胞周期蛋白D1(CyD1)和E2F1的表达及调控情况。我们对4例CD患者和2例健康对照者的结肠外植体进行了体外培养,并用大肠杆菌脂多糖(EC-LPS)进行刺激。用于抑制CyD1和E2F1的商业小干扰RNA(siRNA)被包裹在Invivofectamine®和特意制备的纳米脂质体载体中,以沉默CyD1和E2F1的表达。采用蛋白质免疫印迹分析来研究siRNA对CyD1、E2F1和环氧合酶2(COX-2)表达的影响。在CD患者的结肠外植体中,炎症刺激后CyD1和E2F1表达增加,但siRNA沉默减弱了它们的表达,同时也控制了COX-2的表达。这些数据代表了对体外使用siRNA的初步探索。