CAJ | 학술논문

目的：：获取肝癌相关基因(hepatoma associated gene，HTA)的mRNA分子全长序列并对其可变剪接进行分析，检测其转录本在各肝癌细胞系中的表达，为进一步研究该基因在肝癌发生、发展中的作用奠定基础。方法：用3'RACE和5'RACE方法扩增HTA基因的全长序列并对其序列信息和可变剪接进行扩增和测序分析，用Northern印迹检测该基因转录本在不同肝癌和正常肝细胞系中的表达。结果：HTA基因全长序列为1414 bp，经分析该序列包含3个外显子，2个内含子，其中2号内含子在可变剪接中被作为外显子表达。Northern印迹显示HTA基因1.7 kb转录本和1.4 kb转录本均表达于恶性肝癌细胞系，而不表达于正常细胞系。结论：成功获得了HTA基因的全长序列并对其可变剪接进行了分析，2个大小不同的转录本均在肝癌细胞系中特异性表达，作为一个肝癌相关基因值得进一步深入研究。
목적：：획취간암상관기인(hepatoma associated gene，HTA)적mRNA분자전장서렬병대기가변전접진행분석，검측기전록본재각간암세포계중적표체，위진일보연구해기인재간암발생、발전중적작용전정기출。방법：용3'RACE화5'RACE방법확증HTA기인적전장서렬병대기서렬신식화가변전접진행확증화측서분석，용Northern인적검측해기인전록본재불동간암화정상간세포계중적표체。결과：HTA기인전장서렬위1414 bp，경분석해서렬포함3개외현자，2개내함자，기중2호내함자재가변전접중피작위외현자표체。Northern인적현시HTA기인1.7 kb전록본화1.4 kb전록본균표체우악성간암세포계，이불표체우정상세포계。결론：성공획득료HTA기인적전장서렬병대기가변전접진행료분석，2개대소불동적전록본균재간암세포계중특이성표체，작위일개간암상관기인치득진일보심입연구。
Objective:To obtain the full length cDNA sequences of hepatoma associated gene HTA, analyze its alternative splicing, detect the expression pattern of 2 HTA gene transcripts in different hepatic cell lines, and to establish a base for further study of HTA gene function in hepatocellular carcinoma (HCC) occurence and development. Methods:The full length cDNA of HTA gene was cloned by rapid amplification of cDNA 3' ends (3'-RACE), rapid ampliifcation of cDNA 5' ends (5'-RACE) and DNA sequencing. The gene structure and alternative splicing were analysed. Northern blot assay was performed to detect the expression pattern of 2 HTA gene transcripts in different hepatic cell lines. Results:The full length of HTA gene was 1414 bp, composed of 3 exons and 2 introns, and the second intron could be retained in mRNA. Northern blot assay showed that 2 transcripts of HTA mRNA(1.4 kb and 1.7 kb) could express in the HCC cell lines HepG2 and QGY-7703, but not in the non-malignant cell line L-02 and HUVEC. The expression level of 1.4 kb transcript was much higher than 1.7 kb one. Conclusion:This study successfully has obtained the full length cDNA of HTA gene, and analysed the gene sequence and alternative splicing, 2 transcripts of HTA mRNA specifically expressed in HCC cell lines. As a hepatoma associated gene, HTA deserves further investigation.