Relation between transfection of exogenous CYR61 and expression of integrin / 上海第二医科大学学报

ABSTRACT

Objective To study the induction of integrin molecules mediated by CYR61 in human peripheral blood mononuclear cells(PBMC). Methods A recombinant expression plasmid containing full length of human CYR61 labeled with human IgG Fc fragment was constructed and identified by DNA sequencing.COS7 was used as host cell for identification of secretary CYR61 expression,confirmed by Western blot method.The commercial lipofectin was adopted for recombinant plasmid transfection into PBMC.Real-time PCR was ultilized to analyze expression patterns of CYR61 and integrin molecules in transfected PBMC. Results The insert sequence was correct in the recombinant plasmid.Western blot test showed that CYR61 protein secreted into culture supernatant or was in COS7 cytoplasm.The recombinant plasmid was transfected into PBMC stimulated with phytohemagglutinine(PHA) to induce CYR61 expression and secretion.The results demonstrated that exogenous CYR61 transcribed rapidly after being incubated with PHA and reached the peak after 24 h.But the expression dropped down quickly to a very low level after 48 h.Simultaneously,integrin molecules expressed just after CYR61 transcription.In the set of integrin molecules tested in the study,?v,?M,?3 and ?5 expression were higher than the other integrin molecules(P