mRNA差异显示法筛选大肠癌相关基因

Screening of colorectal carcinoma-related gene by mRNA differential display

目的 分离并克隆大肠癌相关基因。方法分别提取大肠癌与正常大肠粘膜的组织总ＲＮＡ，用ｍＲＮＡ差异显示法 （Differential display polymerase chain reaction,ＤＤ－ＰＣＲ）筛选大肠癌特异性表达产物的ｃＤＮＡ片段，对其进行克隆、测 序、序列分析以及斑点杂交初步鉴定。结果获得了２个在大肠癌高表达而在正常大肠粘膜组织低表达的新基因片段。 结论这２条差异显示新基因片段很可能是与大肠癌相关的致癌基因。

Objective To isolate and clone colorectal carcinoma-related gene. Methods The total RNA was extracted from the tissues of human colorectal carcinoma and normal colorectal mucosa, and the specifically expressed cDNA fragments of colorectal carcinoma were screened by way of differential display polymerase chain reaction (DD-PCR). The cDNA fragments yielded from the above procedures were cloned into the plasmid TA, and subsequently underwent sequencing analysis and dot blot hybridization. Results Two novel cDNA fragments were obtained which were highly expressed in the colorectal carcinoma tissues but not in normal colorectal mucosa. Conclusion These 2 specifically expressed cDNA fragments isolated by DD-PCR might be the colorectal carcinoma-related oncogenes in quest.