应用AFLP法检测小鼠高、低转移性肝癌细胞株基因组的遗传差异  

Application of amplified fragment length polymorphism on detecting genomic alteration of high and low metastatic murine hepatocarcinoma

作　　者：梁智勇[1] 史景泉[1] 魏泓[2] 吴丰春[2] 张树辉[1] 

机构地区：[1]第三军医大学附属西南医院病理学研究所,重庆400038 [2]第三军医大学实验动物中心,重庆400038

出　　处：《第三军医大学学报》2002年第12期1416-1419,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 396 0 0 0 79;3990 0 173) ;全军"九五"医药卫生科研基金资助项目 ( 96M0 83) ;重庆市攻关项目

摘　　要：目的　应用AFLP技术检测小鼠高、低转移性肝癌细胞株基因组的遗传改变 ,以期发现与肿瘤转移相关的DNA片段或基因。方法　肿瘤组织基因组DNA经限制性内切酶酶切 ,连接特异性接头 ,采用 2 5对与接头相识别的引物进行扩增 ,扩增产物经含溴化乙锭的 1 5 %琼脂糖凝胶电泳 ,紫外透视仪分析、照相 ,回收差异片段 ,克隆测序。结果　 2 5对引物均扩增出清晰条带 ,2 1对引物扩增结果显示高低转移性肝癌无差异 ,4对引物扩增结果显示二者之间有差异 ,表现为扩增带的有无 ,4条回收片段中的 1条片段克隆成功并测序。结论　AFLP技术适用于肿瘤易感基因的筛选 ,小鼠肝癌的转移可能与基因组不稳定性有关 ,所得差异片段是否与转移相关有待进一步鉴定。Objective To detect the genetic genomic variant in high and low metastatic murine hepatocarcinoma cell lines for finding special DNA or gene fragment Method Genomic DNAs from tumor tissue were digested by restriction endonuclease, ligated with specific adapter, and amplified by using 25 pairs of primers, PCR products were electrophoretically separated in agarose gel and banding profiles were visualized by ethidium bromide staining,differential bands collected were cloned into pUCm T vector and squenced Results 21 of the 25 pairs of primers produced amplification products were monomorphic and the remaining revealed changes when compared AFLP profiles of two cell lines 4 differential DNA bands were collected and 1 band were successfully cloned and squenced Conclusion AFLP method may be a better technique for finding difference in tumor genome,metastatic ability of murine hepatocarcinoma may be associated with genetic genomic instability,the value of the differential band deserved for further studying

关 键 词：AFLP法 检测 小鼠 转移性肝癌 细胞株基因组 遗传差异 

分 类 号：R735.7[医药卫生—肿瘤]