Secretory Expression and Biological Function of IGFBP-3 in Eukaryotic Cells

Abstract:

The cDNA of Insulin-like growth factor binding protein 3 was subcloned into a eukaryotic secretory expression vector pSectagA to construct pSectag-IGFBP3. Human renal cell carcinoma (RCC) 786-O cells were transfected with pSectag-IGFBP3 using lipofectamine 2000. After 48 h, the secretory IGFBP-3 was tested and identified by western blotting. Meanwhile, Annexin V-EGFP stain was used to analyze the apoptosis of 786-O cells induced by IGFBP-3. Secretory IGFBP-3 protein could express successfully in the 786-O cells and the expressed IGFBP-3 directly displayed an apoptotic effect on the host cells. This work provides a basis for further study on the apoptosis-inducing mechanism of IGFBP-3 and the development of a new anti-tumor drug.

Key words:IGFBP-3, eukaryotic secretory expression, human renal cell carcinoma cells, inducing apoptosis.