βig-h_3基因的真核表达及其对人7721肝癌细胞基质金属蛋白酶表达水平的影响  

Eukaryotic Expression of βig-h_3 Gene and Its Effects on Secretion of MMPs in the Human Hepatoma 7721 Cells

作　　者：唐娟[1] 蒋建利[1] 周宏伟[1] 熊华[1] 杨向民[1] 陈志南[1] 

机构地区：[1]第四军医大学基础部细胞工程研究中心,西安710032

出　　处：《中国生物工程杂志》2006年第9期1-4,共4页China Biotechnology

基　　金：国家自然科学基金资助项目(30200144)

摘　　要：目的:构建βig-h3基因真核表达载体pEGFP-C2/βig-h3并转染人7721肝癌细胞,检测转染后细胞MMPs表达水平的变化。方法:用RT-PCR方法获得βig-h3基因,以pEGFP-C2为载体,构建重组表达质粒pEGFP-C2/βig-h3。重组质粒用脂质体转染人7721肝癌细胞,明胶酶谱法检测转染后细胞MMPs表达水平的变化。结果:正确构建了pEGFP-C2/βig-h3重组质粒,并且在人7721肝癌细胞达到高转染效率,转染后细胞MMPs表达水平明显升高。结论:成功构建了pEGFP-C2/βig-h3真核表达质粒,βig-h3促进人7721肝癌细胞分泌MMPs,提示βig-h3与肝癌的侵袭和转移密切相关。βig-h3 was first identified as a transforming growth factor-betal-inducible gene in human lung adenocarcinoma cell line. It encodes for a secreted extracellular matrix （ECM） protein, which is thought to act on cell attachment and ECM composition. Previous study showed that βig-h3 were highly expressed in human hepatoma cell lines and lowly expressed in human normal hepatic cells. The present study aimed to transfect βig-h3 Matrix metalloproteinase Hepatoma cell Eukaryotic expressionig- h3 into 7721 cells to investigate its effect on secretion of MMPs in the transfected human hepatoma cells. Full- length βig-h3 Matrix metalloproteinase Hepatoma cell Eukaryotic expressionig-h3 gene, cloned by reverse transcription polymerase chain reaction （RT-PCR） was inserted into the eukaryotic expression vector pEGFP-C2. The recombinant plasmid was transfected into 7721 cells with Lipofectamine2000 and Gelatin-Zymography were adopted to detect the production of MMPs in the transfected cells. Results showed that βig-h3 Matrix metalloproteinase Hepatoma cell Eukaryotic expressionig-ha/pEGFP-C2 recombinant expression plasmid was successfully constructed and achieved high transfection efficiency. MMPs expression of the transfected cells was promoted significantly. These results suggest that overexpression of βig-h3 Matrix metalloproteinase Hepatoma cell Eukaryotic expressionig-h3 promoted the production of MMPs, indicating that βig-h3 Matrix metalloproteinase Hepatoma cell Eukaryotic expressionig-h3 may play roles in the invasive and metastatic processes of hepatoma.

关 键 词：βig-h 基质金属蛋白酶 肝癌细胞 真核表达 

分 类 号：Q786[生物学—分子生物学]