Construction of 11β-HSD2 recombinant eukaryotic vector and its effect on IL-6 in A549 cells

Abstract

Objective To establish recombinant eukaryotic vector of 11β-hydroxysteroid dehydrogenase 2 gene in mammalian cell and to detect the expression of interleukin-6 after the vector transfection.Methods The cDNA of 11β-hydroxysteroid dehydrogenase 2 gene was amplified by PCR.After subcloned with pGMT vector and sequenced,the gene was inserted into the eukaryotic vector pcDNA3.1myc hisC.The expression vector was transiently transfected in human lung adenocarcinoma cell line A549 cells,and then the protein was detected using Western blot,interleukin-6 was measured by ELISA.Results Sequencing revealed 11β-hydroxysteroid dehydrogenase 2 recombinant eukaryotic expression vector was constructed successfully,and Western blot indicated that eukaryotic expression vector expressed successfully in A549 cells.Transfection with recombinant eukaryotic vector of 11β hydroxysteroid dehydrogenase 2 gene significantly increased interleukin 6 expression.Conclusions The recombinant eukaryotic vector of 11β-hydroxysteroid dehydrogenase 2 gene is established successfully and could be highly overexpressed in A549 cells.The increase level of interleukin-6 suggests the proinflammatory effect of 11β-hydroxysteroid dehydrogenase 2. Key words: 11β-hydroxysteroid dehydrogenase 2 ; Gene expression ; A549 cells ; Interleukin 6