论文部分内容阅读
目的 :克隆在肝癌组织中低表达的差异显示片段L2的基因cDNA序列 ,并初步探讨其在肝癌发生、发展过程中的作用。方法 :将L2在GenBank中拼接后 ,分析其序列结构 ,并构建正义真核表达质粒 ,转染BEL 740 2肝癌细胞后 ,对转染细胞增殖、粘附、迁移和侵袭等指标进行测定。结果 :克隆得到一条 10 0 5bp的cDNA ,其 5′端 712bp与人Liprinβ1cDNA 5′端调控序列和外显子 1,2 ,3序列同源 (10 0 % ) ,而 3′端 2 93bp与Liprinβ1基因内含子 4同源 (10 0 % )。其间有一个 5 10bp开放阅读框架 ,起始密码位置与Liprinβ1基因相同 ,推断的蛋白产物除C端 13个氨基酸残基外 ,其余均与Liprinβ1同源。此cDNA暂命名为LHS (Liprinβ1 homologoussequence)。LHScDNA正义转染BEL 740 2肝癌细胞系 ,对于细胞的增殖活性和侵袭能力无明显影响 ,但可降低肝癌细胞在层粘连蛋白上的粘附和迁移能力。结论 :LHS表达下调可能在肝癌的转移过程中起重要作用
Objective : Clone the differentially expressed cDNA sequence of L2 gene in HCC tissue and preliminary explore its role in the development and progression of HCC. METHODS: After L2 was ligated in GenBank, its sequence structure was analyzed and a sense eukaryotic expression plasmid was constructed. After transfection of BEL 740 2 hepatoma cells, the proliferation, adhesion, migration, and invasion of transfected cells were measured. RESULTS: A 10 5 bp cDNA was cloned and its 5’ end 712 bp was homologous to the 5’-end sequence of human Liprin β 1 cDNA and exon 1, 2, 3 (10 0 %), while the 3’-end was 2 93 bp and Liprin β 1 Gene intron 4 is homologous (100%). In the meantime, there is a 5-10bp open reading frame, and the start position is the same as that of Liprinβ1 gene. The deduced protein product is identical to Liprinβ1 except for 13 amino acids at the C-terminus. This cDNA is provisionally named LHS (Liprinβ1 homologous sequence). LHS cDNA sense transfection of BEL 740 2 liver cancer cell line had no significant effect on the cell proliferation activity and invasion ability, but it could reduce the adhesion and migration ability of hepatoma cells on laminin. Conclusion: Down-regulation of LHS may play an important role in the metastasis of liver cancer