【作者】 周清； 郭爱林； 安社娟； 徐崇锐； 王震； 吴一龙；

【Author】 Department of lung neoplasms, Guangdong Provincial People’ s Hospital, Guangzhou 510080, China

【机构】 广东省人民医院肿瘤中心肺科；

【摘要】 目的:探讨XAGE-1b基因在非小细胞肺癌中的表达及与临床特点的相关性。方法:经组织学或细胞学证实的非小细胞肺癌患者30例,通过手术获得每例患者的肺癌组织和癌旁正常肺组织,切成大小约0．2～0．3克的组织块,快速将组织块置入无菌,去RNA酶的冻存管,投入液氮,随后转入-80℃低温冰箱保存,采用GBICOL BRL公司一步法提取肺癌组织和癌旁正常肺组织的总RNA,采用紫外分光光度计法和琼脂糖凝胶电泳法鉴定总RNA质量,软件设计扩增 XAGE-1b基因全长的引物序列,以GAPDH基因为反应体系内参照进行RT-PCR反应,通过琼脂糖凝胶电泳和基因测序鉴定RT—PCR产物,确定为基因表达阳性标本,采用 SPSS10．0统计软件对基因表达阳性率及与临床特征相关性进行分析。结果:从每例患者0．2～0．3克肺癌组织或正常肺组织中可以提取100～300 μg RNA,OD260／OD280比值在 1．8～2．0,电泳显示28S,18S条带清晰,28S条带亮度约为18S的1．5倍,30例肺癌组织中,XAGE-1b基因表达阳性率为40％(12／30),30例癌旁正常肺组织不表达该基因,基因表达与患者年龄,性别,肿瘤分化程度无相关性, 与病理类型有相关性, 腺癌中基因表达阳性率为61．1％ (11／18),明显高于其他病理类型的8．3％(1／12)(P=0． 015),随着TNM分期的升高,基因表达阳性率略有增加, 但差异无统计学意义(P>0．05)。结论:XAGE-1b基因在非小细胞肺癌中有较高的表达阳性率,癌旁正常肺组织不表达该基因,是较理想的肺癌特异性免疫治疗靶点,尤其在肺腺癌中可以优先选择该基因作为免疫治疗靶点。更多还原

【Abstract】 Objective: To explore the expression of XAGE-1b gene in non-small cell lung cancer and the relationship between gene expression and clinical characteristics. Methods: 30 cases with resectable non-small cell lung cancer were chosen. After pulmonary lobectomy, tumor tissues and normal lung tissues were collected and put into liquid nitrogen immediately and then were stored in -80 ℃ refrigerator. Total RNA from tumor tissue and normal lung tissue was extracted from the tissues by using a standard TRIZOL protocol. Total RNA quality was identified by using ultraviolet spectrophotom-eter and agarose gel electrophoresis. Primers were designed to amplify full-length gene. RT-PCR were carried out by using RT-PCR kit. The products of RT-PCR were identified by electrophoresis and sequencing. SPSS 10.0 software was used to analyze the expression of XAGE-lb gene and relationship with clinical characteristics in non-small cell lung cancer. Results: 100 - 300 μ g RNA was extracted from 0.2 - 0.3 gram tissue and the value of OD260/OD280 was 1.8 - 2.0. Electrophoresis showed that 28S and 18S were clear. In 30 cases, expression rate of XAGE-1b was 40%. Normal lung tissues did not express this gene. Gene expression had no relationship with age, gender, tumor differentiation or clinical stages, but had relationship with pathology. Expression rate of XAGE-1b in adencarcinoma ( 61.1% ) was much higher than in other pathology ( 8. 3% ) ( P=0.015 ). There was a increase tendency of gene expression with the up-stage of TNM but there was no significant difference ( P>0.05 ). Conclusion: XAGE-1b gene expresses highly in non-small cell lung cancer tissues but none in normal lung tissues, which makes it to be an ideal target of immunotherapy, especially in adenocarci-noma of lung.更多还原