FasL基因的克隆化及其mRNA在肺癌组织中的表达  

Clone of Fasligand gene and its expression of mRNA in lung cancer

作　　者：沈毅[1] 彭传亮[1] 罗宜人[1] 魏煜程 吕振华[1] 

机构地区：[1]青岛大学医学院附属医院,山东青岛266003

出　　处：《山东医药》2003年第9期7-8,共2页Shandong Medical Journal

摘　　要：用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经病理证实的肺癌及癌周正常肺组织新鲜标本 ,用液氮速冻 ,4%多聚甲醛固定。在冰冻切片上以 c RNA探针原位杂交进行 Fas L m RNA表达分布的观察。结果 :在鳞癌、腺癌和小细胞肺癌的癌组织及癌旁肺组织中均可检出 Fas L的杂交信号。镜下见鳞癌组织中 FasHuman PBMC was treated by PHA-P to induce expression of FasL mRNA,total RNA was extracted from the actived PBMC,FasL gene fragment was selected through RT-PCR.The gene was cloned into pSPT 19 in multi-cloning site to acquire recombinant plasmid,and was identified by DNA sequencing,then Dig-FasL cRNA probe was synthesized in vitro.Lung cancer samples and normal lung tissures near the tumor were collected,and were snap-frozen tith liguid nitrogen,fixed with 4%PFA.The distribution of FasL mRNA was observed in lung cancer tissues with CRNA probe in situ hybridization on frozen section.The results showed that the FasL mRNA signals emerged in squamous,adenocarcinoma and small cell lung cancer cells.The FasL were expressed more highly in squamous cells than in adenocarcinoma and small cell lung cancer cells.

关 键 词：FASL基因 克隆化 MRNA 肺癌组织 CRNA探针 原位杂交 

分 类 号：R734.2[医药卫生—肿瘤]