Lentivirus-mediated cyclooxygenase 2 and aggrecanase 1 silencing and insulin-like growth factor 1 overexpression in human bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage. OBJECTIVE:To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cels. METHODS:Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cels culturedin vitro (experimental group). The human bone marrow mesenchymal stem cels transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cels transfected with no treatment served as blank group. RESULTS AND CONCLUSION:Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cels were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfuly silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cels, which brings a new hope for the systemic gene treatment of arthritis.