信号传导及激活因子2基因在食管癌中表达.docVIP

信号传导及激活因子2基因在食管癌中表达 　【摘要】 目的： 筛选食管癌中特异表达的基因，进一步了解食管癌发生的分子机制。方法： 利用荧光差异显示技术(DD-PCR)获得差异表达的片段，对这些片段进行克隆和序列分析。通过在基因库中同源性检索，查找差异片段相对应的同源基因。利用实时荧光定量PCR技术在食管癌临床标本中进行验证。结果： 通过DD-PCR获得的差异片段中有一个片段对应于信号传导与激活因子2(stat2)基因，该基因的读码框长2 556 bp，编码852个氨基酸，编码蛋白分子量约97.9 kD。应用实时荧光定量PCR方法进一步验证发现该基因在食管癌组织中的表达量高于其对应的癌旁组织（Plt;0.05, n=16）。结论： stat2基因在食管癌组织中呈高表达，推测其可能在食管癌的发生发展中起一定的作用。 【关键词】 荧光差异显示; 定量PCR; 食管癌; 信号传导与激活因子2; 基因表达 　［Abstract］ Objective: To further investigate the molecular mechanism of esophageal carcinogenesis, and to validate the genes expressed differently in esophageal cancer. Methods： mRNA differential display(DD-PCR) was employed to search differently expressed fragments, some of which were cloned and sequenced. By homology analysis in GenBank, corresponding homologous genes of those fragments were found. The corresponding genes of those differently expressed fragments were validated by Real-time quantitative PCR. Results： By DD-PCR, one of the highly expressed EST in cancer tissues was identified to be Homo sapiens signal transducer and activator of transcription 2(stat2)gene. The results of real-time quantitative PCR showed that the expression level of stat2 in esophageal cancer tissues were significantly higher than that in normal tissues(n=16, Plt;0.01). The ORF of stat2 gene was 2556 base pair long and encodes 852 amino acids. The molecular weight was about 97.9 kD. Conclusion： The over-expression of stat2 gene may indicate an important role in the occurrence and development of esophageal cancer. 　［Key words］ fluorescent differential display; real-time quantitative PCR; esophageal cancer; stat2; gene expression 　　食管癌是最主要的恶性肿瘤类型之一，我国是世界上食管癌的高发地区。同时，食管癌的发生在我国也具有地区差异性，如河南省林县和江苏省扬中地区等都是食管癌高发地区。目前，已发现一些食管癌的癌变机制相关因素，但关键原因尚未明确［1］。癌基因、抑癌基因和细胞周期控制基因的异常表达参与了食管癌及其他恶性肿瘤的演进［2］,如Ohta等［3］的研究表明肿瘤抑制基因-FHIT基因，在大约50%的食管癌、胃癌和结肠癌中存在转录异常，Gleeson等［4］在Barrett食管腺癌中发现了特异性p53突变谱。而细胞周期控制基因cyclinD1的扩增与食管鳞癌明显相关［5］。在癌基因中stat(singnal transducers and transcription activator， stat)家族和myc家族等均对肿瘤的发生起重要作用