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目的筛选和克隆结肠癌和正常结肠组织差异表达的基因片段,为探讨结肠癌的发病机制提供线索。方法应用基因差异显示技术(DD-PCR),比较结肠癌和正常结肠组织基因表达的差异,对其中一条有明显差异的基因片段进行克隆、测序,测序结果提交GenBank数据库中进行同源性分析,并用半定量RT-PCR进行初步鉴定。结果同源性分析表明,该片段与已知基因DDX32高度同源(99%)。RT-PCR结果显示,在结肠癌组织中该基因mRNA的表达水平显著高于正常结肠组织(P<0.05)。结论DD-PCR是筛选差异表达基因的有效手段;在结肠癌组织中DDX32基因的表达显著高于正常结肠组织,此结果为研究DDX32基因在结肠癌发病中的作用提供了线索。
Objective To screen and clone gene fragments differentially expressed in colon and normal colon tissues and provide clues for exploring the pathogenesis of colon cancer. Methods The gene expression differences between colon cancer and normal colon tissues were compared by DD-PCR. One of the differentially expressed genes was cloned and sequenced. The sequencing results were submitted to GenBank for homology analysis. Semi-quantitative RT-PCR was used for preliminary identification. Results Homology analysis showed that the fragment was highly homologous (99%) with the known gene DDX32. RT-PCR results showed that the mRNA expression of this gene in colon cancer tissues was significantly higher than that in normal colon tissues (P <0.05). Conclusion DD-PCR is an effective method to screen differentially expressed genes. The expression of DDX32 gene in colon cancer tissues is significantly higher than that in normal colon tissues. This result provides a clue to study the role of DDX32 gene in the pathogenesis of colon cancer.