臆断引物-聚合酶链反应分析配对胃腺癌组织与非癌胃组织基因组的差异基因

A STUDY ON THE GENOMIC VARIANT IN MATCHED ADENOCINOMA AND NON-TUMOR GASTRIC TISSUE BY ARBITRARILY PRIMER POLYMERASE CHAIN REACTION

目的 研究同一胃腺癌患者的配对胃腺癌组织与非癌胃组织基因差异 ,并对差异基因片段进行克隆 ,Southern印迹杂交。方法 应用随意扩增多态性DNA指纹图谱分析技术 (arbitrarilyprimerpolymerasechainreaction ,AP PCR) ,即臆断引物的无细胞分子克隆技术 ,亦称臆断引物的聚合酶链反应 ,对 5例配对标本作基因组差异分析。结果 与各自配对的非癌胃组织相比 ,所有胃腺癌组织基因组DNA的AP PCR指纹图谱均存在变异 ,并克隆了其中 1例仅存在于肿瘤组织基因组的差异扩增片段PW 2 .2。Southern印迹杂交发现此PW 2 .2也存在于其他部分胃腺癌标本的随机扩增产物中。结论 AP PCR技术可为差异基因分析和克隆提供一快速而有效的方法。

Objective To identify and isolate the variant gene associated with gastric adenocarcinoma and clone the fragment of variant gene.Methods By arbitrarily primer polymerase chain reaction (AP-PCR), DNA samples from 5 matched gastric adenocarcinoma and non tumor gastric tissues were analysed. Results The produced AP-PCR profiles were different in each matched gastric adenocarcinoma and non-tumor gastric tissue. One differentiated amplified DNA fragments PW2.2 from a matched gastric adenocarcinoma were cloned. The result of Southern blot hybridization with PW2.2 as a probe showing that this fragment was also found in some other gastric adenocarcinoma samples. Conclusion AP-PCR fingerprinting assay can be used to identify and clone the variant genes associated with gastric adenocarcinoma.