bcl-2家族相关基因异常表达与Barrett食管的关系

Abnormal expression of bcl-2 gene family in development of Barrett＇ s esophagus

目的应用寡核苷酸芯片技术高通量分析Barrett食管与正常食管黏膜组织的基因表达差异，探索与疾病进展相关的靶基因。方法对Barrett食管患者的病变食管黏膜以及正常食管黏膜组织，应用Trizol一步法进行总RNA抽提，10g/L琼脂糖凝胶电泳和芯片实验室进行RNA质量检测。总RNA纯化后进行逆转录cRNA合成、荧光标记和纯化，将Barrett食管黏膜和正常食管黏膜组织的cRNA探针分别与Agilent寡核苷酸芯片（30，968探针）进行杂交，洗涤后应用Agilent扫描仪获取图像，Feature Extraction提取软件进行定量分析处理。结果（1）大活检钳钳取2次活检的组织能提供芯片所需要的5μg RNA，配对组织总RNA质量高，反转录cRNA及荧光标记质量好；（2）2倍差异表达基因中，上调基因共142个，下调基因共284个。其中包括bcl-2、MCLI、BAX、BIK和BCLAF1等15个异常表达的bcl-2家族相关基因。结论基因芯片分析可用于内镜下活检组织，Barrett食管的发生发展涉及多基因多步骤，是一个复杂的过程；bcl-2家族相关基因异常表达可能涉及Barrett食管的发生发展过程，确切机制有待进一步深入研究。

Objective To detect the differential expression genes （DEGs） between Barrett＇s esophagus （BE） and normal esophagus with oligomicroarray, and to explore the target genes related to the development of BE. Methods The total RNAs of matched BE and normal esophagus mucosa from same patient were isolated with one step Trizol method. Matched RNAs were qualified with 10g/L agarose gel electrophorests. After tRNA purification, cRNAs were synthesized and labeled with fluorescence, which were then hybridized with Agilent oligomicroarray containing 30,968 probes. The fluorescence intensity features were detected by Agilent scanner and quantified by software Feature Extraction. Results On average, 2 biopsies by disposable jumbo biopsy forceps provided approximately 5 μg RNA required for microarray. The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality. Among 2-fold DEGs, there were 142 up-regulated genes and 284 down-regulated genes including 15 bcl-2 related genes such as bcl-2, MCL1, BAX, BIK and BCLAF1. Conclusion Microarray-based studies are feasible in endoscopieally obtained tissues. The development of BE is a complicated process involving multi-genes, in which abnormal expression of bcl-2 family related genes might be involved, but the exact mechanism needs further research.