川北地区胃癌特异性表达基因的克隆(1)
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[摘要] 目的：构建胃低分化腺癌组织cDNA消减文库，筛选胃癌特异表达基因。方法：取胃低分化腺癌组织和癌旁正常黏膜组织，提取总RNA，逆转录cDNA，进行SSH构建消减文库获得序列标签，与载体连接、克隆、筛选、测序及同源性检索。结果：以癌旁正常黏膜组织为driver、胃低分化腺癌组织为tester成功构建cDNA消减文库，测序结果与公共数据库中已知基因和人类EST比较，获得5个特异性序列标签(NBPF1、FAM48A、RXFP2、ACTB和QDPR)。结论：应用SSH法成功构建胃低分化腺癌组织cDNA消减文库，初步筛选胃癌部分表达基因，为寻找川北地区胃癌发生发展的特异性基因奠定基础。
[关键词] 胃癌 SSH 特异基因
[中图分类号] R735.2 [文献标识码]A [文章编号]1673-7210(2011)02(b)-019-03
Cloning associated special genes in gastric cancer of North Sichuan Area
WANG Chaoli, LI Li, CHEN Guo, YANG Shangjun, JING Baoqian*, REN Bixuan, FENG Li
(Molecular biology research institute of North Sichuan Medical College, Nanchong 637000, China)
[Abstract] Objective: To construct cDNA subtracted libraries from gastric poorly differentiated adenocarcinoma and screen differentially expressed genes. Methods: Relatively pure gastric poorly differentiated adenocarcinoma and normal gastric mucous membrance were procured, and abstracted the total RNA, and the cDNA was been reversed transcription, which was used to carry on for suppression subtractive hybridization(SSH). Subtracted cDNA fragments were linked with vector, cloned, screened, sequenced, and made homologous search. Results: The subtracted cDNA libraries were constructed with the tester of gastric poorly differentiated adenocarcinoma and the driver of normal mucosa tissue beside of the dysplasia.The sequenced result compared with the known genes of PubMed and human EST and five specificity sequences were procured (NBPF1, FAM48A, RXFP2, ACTB and QDPR). Conclusion: Subtracted cDNA libraries from gastric poorly differentiated adenocarcinoma using SSH. Some fragments have been screened and verified to help to search for novel associated genes with gastric cancer of North Sichuan Area.
[Key words] Gastric cancer; Suppression subtractive hybridization; Special gene
胃癌是严重危害人类健康和生命的恶性肿瘤之一，我国每年有20余万例新发胃癌患者，占全部恶性肿瘤患者的17.2%，其发病率居所有恶性肿瘤发病率之首。每年胃癌患者死亡人数占所有癌症死亡人数的23.93%。由于胃癌起病隐匿，发生的确切机制至今尚不清楚，临床早期70%以上毫无症状，中晚期才引起相应的临床症状，且易转移与复发，预后极差。目前最为紧迫的是寻找对胃癌检测灵敏度高、特异性好的检测方法，使其能对胃癌高危险患者进行筛查、对早期胃癌能特异诊断、对病情发展和疗效可追踪考核，从而提前对高危患者给予干预治疗 ......
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