Macpherson A J, Chester K A, Robson L, Bjarnason I, Malcolm A D, Peters T J
Department of Clinical Biochemistry, Kings College School of Medicine and Dentistry, London.
Gut. 1992 May;33(5):651-6. doi: 10.1136/gut.33.5.651.
The steady state levels of c-myc mRNA have been measured in RNA samples extracted from colonoscopic biopsies of inflammatory bowel disease patients obtained at routine endoscopy sessions. Biopsies were immediately frozen in liquid nitrogen limiting the ischaemic time to less than 15 seconds, and can be stored for up to 96 hours before separation of RNA. Yields of RNA using biopsies were 0.137 (0.041)% wet wt (mean (SD), n = 68), these are significantly better than those obtained from surgical material (0.064 (0.063)% wet wt (mean (SD), n = 21) where the tissue ischaemic time was 45 minutes to one hour 40 minutes. Functional activity of RNA extracted was demonstrated by the ability to direct in vitro protein translation in the rabbit reticulocyte system. We have used this technique to show that there is an increased ratio of steady state c-myc proto-oncogene expression in inflamed tissue from 18 patients with left sided ulcerative colitis and five patients with segmental Crohn's colitis, compared with an uninvolved region of the colon in each case. No difference in c-myc expression was seen in biopsies at least 30 cm apart in 11 control patients with no macroscopic or histological abnormalities. Increased expression of c-myc in inflammatory bowel disease is consistent with the activation of this proto-oncogene during altered cell cycle control resulting from the inflammatory process.
已在从炎症性肠病患者结肠镜活检中提取的RNA样本中测量了c-myc mRNA的稳态水平。这些活检样本在液氮中立即冷冻,将缺血时间限制在15秒以内,并且在分离RNA之前可保存长达96小时。使用活检样本获得的RNA产量为0.137(0.041)%湿重(平均值(标准差),n = 68),这些产量显著高于从手术材料中获得的产量(0.064(0.063)%湿重(平均值(标准差),n = 21),手术材料的组织缺血时间为45分钟至1小时40分钟。提取的RNA的功能活性通过在兔网织红细胞系统中指导体外蛋白质翻译的能力得以证明。我们使用该技术表明,与每种情况下结肠的未受累区域相比,18例左侧溃疡性结肠炎患者和5例节段性克罗恩结肠炎患者炎症组织中稳态c-myc原癌基因表达的比例增加。在11例无宏观或组织学异常的对照患者中,相距至少30 cm的活检样本中未观察到c-myc表达的差异。炎症性肠病中c-myc表达的增加与炎症过程导致的细胞周期控制改变期间该原癌基因的激活一致。