Zhang Yuwei, Lai Maode, Gu Xuemei, Luo Minjie, Shao Lina
Department of Pathology, School of Medicine of Zhejiang University, Hangzhou 310006, China.
Chin Med J (Engl). 2003 Jun;116(6):918-22.
To discover and identify differentially expressed genes associated with colorectal adenoma formation and the role of RegIV in colorectal adenoma differentiation.
A subtracted cDNA library was constructed with cDNAs that were isolated from either the normal mucosa or adenoma tissue of a single patient. Suppressive subtractive hybridization (SSH) combined with virtual northern blotting was used to characterize differentially expressed genes and contigs were assembled by electronic cloning (in silico cloning) with the EST database. Semi-quantitative RT-PCR was performed in 9 colorectal adenomas.
The amino acid sequence was determined with open reading frame (ORF) prediction software and was found to be 100% homologous to the protein product of RegIV (a novel gene isolated from a large inflammatory bowel disease library). RegIV was found to be highly expressed in all of the adenoma samples (9/9) compared with the normal mucosa samples, while 5/6 cases showed RegIV to be more strongly expressed in adenocarcinoma.
RegIV may play an important role in the initiation of colorectal adenoma differentiation, and its detection may be useful in the early diagnosis of colorectal adenoma formation.
发现并鉴定与结肠直肠腺瘤形成相关的差异表达基因以及RegIV在结肠直肠腺瘤分化中的作用。
用从一名患者的正常黏膜或腺瘤组织中分离出的cDNA构建消减cDNA文库。采用抑制性消减杂交（SSH）结合虚拟Northern印迹法来鉴定差异表达基因，并通过与EST数据库进行电子克隆（计算机克隆）来组装重叠群。对9个结肠直肠腺瘤进行半定量逆转录聚合酶链反应（RT-PCR）。
用开放阅读框（ORF）预测软件确定了氨基酸序列，发现其与RegIV（从一个大型炎症性肠病文库中分离出的一个新基因）的蛋白质产物具有100%的同源性。与正常黏膜样本相比，发现RegIV在所有腺瘤样本（9/9）中高表达，而在5/6例腺癌中RegIV表达更强。
RegIV可能在结肠直肠腺瘤分化起始中起重要作用，其检测可能有助于结肠直肠腺瘤形成的早期诊断。
