[Effects of human BMP2 gene transfection on NIH3T3 cells in vitro].

OBJECTIVE

To elucidate the mechanism of bone morphogenetic protein (BMP) on cell differentiation and to provide basis for BMP gene therapy.

METHODS

A phagemid expression vector for human BMP2 (pBK-B2) was constructed and transfected into the NIH3T3 cells by using Lipofect AMINE. Positive cell clones were selected with G-418. The stable transfection, expression and secretion of hBMP2 were determined by in situ hybridization, immunohistochemistry and sandwich-in ELISA methods. The proliferativity of the transfected cells were assayed by methabenzthiazuron (MTT) method. Alkaline phosphatase (ALP) and osteocalcin (OC) production were also measured in the transfected cells.

RESULTS

The results showed that cell proliferation was inhibited after the transfection with hBMP2 gene. But the transfected cells showed increased ALP activity and OC production.

CONCLUSION

These results indicate that BMP2 is expressed stably and efficiently in the NIH3T3 cells and is involved in inducing differentiation of NIH3T3 cells into osteoblast-like cells.