摘要
目的寻找食管癌发生的相关基因，为食管癌易感个体的筛查和早期诊断提供分子标志物。方法对来源于食管癌高发区河南省林县的１２例配对食管癌组织和非癌食管组织ＤＮＡ，应用随机引物聚合酶链反应（ＡＰ－ＰＣＲ）技术分离差异ＤＮＡ片段，进行克隆、测序和序列分析。根据序列的酶切位点分析，选择ＤｒａⅠ和ＨｉｎｄⅢ内切酶，对配对的食管癌组织和非癌食管组织ＤＮＡ酶切，Ｓｏｕｔｈｅｒｎ印迹杂交分析。结果１２例配对食管癌组织和非癌食管组织中，３例癌组织有１．０ｋｂ左右的差异随机扩增片段，而相应的非癌组织缺如。其中的１１Ｔ差异ＤＮＡ片断经测序和序列的同源性分析，基因文库内无同源序列。Ｓｏｕｔｈｅｒｎ印迹杂交显示，配对的食管癌组织和非癌食管组织有明显差异。结论１１Ｔ差异ＤＮＡ片段是食管癌变过程中所发生的改变。是否为一个新的候选癌基因或癌基因标志物。
Objective To isolate the genes involved in carcinogenesis of esophageal cancer and to provide a molecular marker for screening and early detection of it in susceptible individuals. Methods DNA extracted from 12 matched pairs of esophageal cancer and non cancerous tissues in patients of Linxian County, Henan Province, a high incidence area for esophageal cancer, and differential DNA fragments were isolated with arbitrary primer polymerase chain reaction (AP PCR), cloned and sequenced. Restriction endonuclease Dra Ⅰ and Hind Ⅲ were used to digest genomic DNA in the pair matched cancerous and non cancerous esophageal tissues for Southern blot analysis. Results There was a randomly amplified differential DNA fragment of about 1.0 kb in cancerous tissues in three of the twelve cases with esophageal cancer, but no such fragments could be found in corresponding non cancerous tissues. Sequence analysis showed that no homologous DNA in GeneBank database as same as 11T differential DNA fragments. Southern blot analysis showed that there was significant difference between pair matched cancerous and non cancerous tissues. Conclusion 11T differential DNA fragment appeared during carcinogenesis of the esophagus, and further investigations were needed to study if it could be regarded as a new kind of oncogene or a biomarker of it.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
1998年第3期168-170,共3页
Chinese Journal of Preventive Medicine
关键词
食管肿瘤
DNA指纹法
聚合酶链反应
克隆
Esophageal neoplasms DNA fingerprinting Polymerase chain reaction
