In Vitro And In Vivo Inhibition Effect Of Noggin Eukaryotic Expression Vector-pcDNA3.1/his-Noggin On The Proliferation Of Osteosarcoma Cells

Objective To test the expression levels of BMP-2 and Noggin in human osteoblastic cells and osteosarcoma cells. Construct and identify a Noggin eukaryotic expression vector and observe its effect on the proliferation of the osteosarcoma cells in vitro and in vivo.Methods Test the expression levels of BMP-2 and Noggin mRNA in human osteoblastic cell line hFOB 1.19 and osteosarcoma cell line MG-63 by RT-PCR method and observe the effect on the proliferation of the osteosarcoma cells after given different dosages of exogenous Noggin proteins. Then construct a Noggin eukaryotic expression vector—pcDNA 3.1/his-Noggin, and confirm it by DNA sequencing. After transfected pcDNA 3.1/his-Noggin vector into the MG-63 cells, the stable expression strain was obtained by G418 selection. The human osteosarcoma cells MG-63, cells transfected with empty vector and cells transfected with pcDNA 3.1/his-Noggin expression vector were tested by MTT method for cell proliferation, and analyzed by flow cytometry for cell cycle distribution. Then the three groups of osteosarcoma cells were seeded subcutaneously into nude mice. The in vivo carcinogenic time, the final weigh and volume of subcutaneous tumors were measured, and the morphological changes of tumor cells were observed under microscope.Results The expression level of BMP-2 mRNA was significant higher in human osteosarcoma cells than that in human osteoblastic cells, and after given the exogenous Noggin protein, the proliferation of the osteosarcoma cells was inhibited in a dose and time dependent fashion. The pcDNA 3.1/his-Noggin expression vector was successfully constructed and the stable strain was obtained. In vitro study showed significant inhibition on the proliferation of the osteosarcoma cells after transfected with pcDNA 3.1/his-Noggin expression vector, and obvious G0/G1 phase arrest occurred in the cell cycle. And in vivo study showed that after transfected with pcDNA 3.1/his-Noggin expression vector, the carcinogenic activity of osteosarcoma cells was significantly decreased, and the final weigh and volume of subcutaneous tumors 21 days after implantation of the tumor cells was also inhibited.Conclusion There're different expression patterns of BMP-2 and Noggin between human osteosarcoma cells and osteoblastic cells, and the proliferation of the osteosarcoma cells can be significantly inhibited by exogenous noggin protein or pcDNA 3.1/his-Noggin expression vector in vitro and in vivo. Thus, Noggin could be one of the possible targets for the gene therapy of the human osteosarcoma.