几种肺癌细胞系中FHIT基因和蛋白表达的研究  

Expression of FHIT Gene in Different Kinds of Human Lung Cancer Cell Lines

作　　者：张立群[1] 汪蕙[1] 赖百塘[1] 张春艳[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所

出　　处：《结核病与胸部肿瘤》2005年第3期165-169,共5页Tuberculosis and Thoracic Tumor

摘　　要：目的研究不同人肺癌细胞系FHIT基因和蛋向的表达，为探讨外源FHIT基因对不同FHIT状态人肺癌细胞恶性表型的影响筛选受体细胞。方法提取B01D（人肺巨细胞癌细胞系）、LTEP-A2（人肺腺癌细胞系）、LTEP-Sm（人小细胞肺癌细胞系）和A549（人肺腺癌细胞系）细胞总RNA，定量后逆转录合成cDNA第一链，分别用5131、3D1和5U2、3D2两对引物进行巢式PCR扩增FHIT基因外显子1～10。1．5％琼脂糖凝胶电泳检测4种细胞株的FHIT基因表达。将扩增的PCR产物纯化后进行DNA测序。用FHIT单克隆抗体进行免疫组化染色检测4种细胞FHIT蛋白表达。结果801D和LTEP-Sm细胞有正常FHIT基因转录本，LTEP-A2细胞有一条正常FHIT基因转录本和一条异常转录本，正常转录本测序显示与FHIT cDNA同源，无缺失及重排。801D和LTEP-Sm细胞FHIT蛋向表达呈强阳性。A2细胞FHIT蛋白表达减弱。A549缺乏FHIT基因转录本和蛋白表达。结论4种细胞系代表3种FHIT基因状态，为进一步研究外源FHIT基因对不同FHIT状态人肺癌细胞恶性表型的影响提供了合适的受体细胞。Objective To examine the expression of FHIT gene and protein in four kinds of human lung cancer cell lines, and to screen the cell lines used for exogenous FHIT gene transfection. Methods The total RNA of 801D ,LTEP-Sm,LTEP-A2 and A549 cell lines were extracted. The first strand eDNA was constructed. The FHIT gene exonl to exonl0 was amplified by nested RT-PCR using the primer 5U1,3D1 and 5U2,3D2 and The PCR products was sequenced.To examine the expression of FHIT protein in 801D ,LTEP-Sm ,LTEP-A2 and A549 cell lines,immunochemical stain was done using the FHIT antibody in these cancer cell lines.Results There was a normal transcription of FHIT gene in 801D and LTEP-Sm cell lines.A normal and an abnormal transcription were found in LTEP-A2 cell lines.No gene deletion and rearrangement was tested in the normal transcription. There was a high level expression of FHIT protein in 801D and LTEP-Sm cell lines. No detectable levels of wild-type FHIT mR-NA transcriipt and protein were presented in the A549 cell lines.Conclusion There are different status of FHIT gene in the different human lung cancer cell lines.The influence of extrogenous FHIT gene on the malignant phenotype of these cell lines can be investigated.

关 键 词：肺肿瘤 FHIT基因 反转录巢式PCR 人肺癌细胞系 蛋白表达 表达的研究 人肺腺癌细胞系 巢式PCR扩增 人肺巨细胞癌细胞系 基因转录本 

分 类 号：R734.2[医药卫生—肿瘤] R730.3[医药卫生—临床医学]