肿瘤相关基因NAG6在胃癌中表达及限制性片段长度多态性分析.docVIP

肿瘤相关基因NAG6在胃癌中表达及限制性片段长度多态性分析 　【摘要】 目的 检测NAG6基因在胃癌及癌旁组织中的表达，探讨此基因在胃癌发生、发展中的作用以及它在胃癌中的基因结构变化。方法 采用RTPCR，Northern杂交，点杂交方法检测42例胃癌和癌旁组织中NAG6的表达；RFLP分析检测该基因在胃癌中是否存在基因结构的改变。结果 59.5%（25/42）的胃癌组织中NAG6基因表达缺失,NAG6在胃癌组织中的表达较癌旁组织显著下调（Plt;0.005）,但NAG6表达下调与胃癌组织类型、淋巴结或远处转移无明显关系（P＞0.05）。点杂交证实NAG6在胃癌组织中表达下调。RFLP分析发现NAG6在7例胃癌及癌旁组织中的基因型为11.5Kb/5.0Kb,在3例癌组织中发现5.0Kb这条等位基因片段的杂合性丢失。结论 NAG6基因在胃癌组织中表达显著下调，该基因的表达下调可能在胃癌的发生和发展过程起到一定的作用，但是与胃癌的转移无明显关系。而遗传物质的丢失可能与该基因表达下调有关。提示NAG6基因可能为定位于7q3132的与胃癌相关的侯选抑瘤基因。 【关键词】 胃癌 抑瘤基因 基因表达 RFLP 　　Expression of Tumor Related Gene NAG6 in Gastric Cancer and RFLP Analysis 　　Abstract：Objective To examine the expression level of NAG6 gene in gastric cancer and matched paracancerous tissues, and to investigate its role in the occurrence and development of gastric cancer, also to study if the genetic structure of NAG6 was alteration in gastric cancer. Methods Reverse transcriptionpolymerase chain reaction (RTPCR), Northern blot analysis and Dot hybridization were used to compare the expression level of NAG6 gene in 42 cases of human gastric cancer tissues with their matched paracancerous tissues of the same patients respectively. In addition, Restriction Fragment Length Polymorphism (RFLP) analysis was adapted to study if the genetic structure of NAG6 was alteration in gastric carcinomas. Results The expression of NAG6 was absent in 57.1% gastric cancer tissues (25/42). The expression of NAG6 was significantly downregulated in gastric cancer tissues compared with that in matched paracancerous tissues (Plt;0.005). However downregulation of NAG6 was not relevant to pathologic type and lymphnode or distance metastasis of gastric cancers（Pgt;0.05）. Dot hybridization confirmed the results of RTPCR. Furthermore, the results of EcoRⅠRFLP analysis of NAG6 gene demonstrated that 3 of 7 cases of gastric cancer showed loss of 5kb fragment in comparison with their matched paracancerous tissues. Conclusion NAG6 gene was significantly downregulated in gastric cancer ti