A study of biological function of retinoblastoma gene (Rb gene)

The human wild-type Rb gene cDNA has been cis- or trans inserted into the retrovirus vector DOL, resulting in a sense-expression vector DOLRB and an antisense-expression vector DOLRBAS of Rb gene. By eletroporation transfection techniques, the vector DOLRB has been introduced into the human breast carcinoma cell line MDAMB468 and human hepatocellular carcinoma cell line SMMC7721 both of which have an inactivated Rb gene and the vector DOLBAS, into normal human embryonic lung fibroblasts HEL cells. With the expression of Rb protein, the growth rate of the MDAMB468 cells is decreased by about 50%, their colony formation ability in soft agar is repressed completely, and their tumorigenicity in nude mice is repressed partially. Meanwhile, the cell population of G1 phase of Rb+ MDAMB468 cells is increased markedly. About 75% of transfected SMMC7721 cells have been killed by Rb gene product. For HEL cells, with the transient expression of antisense Rb gene, the Rb protein synthesis is reduced and the growth rate of those cells increased, but no colonies of HEL cells are formed in soft agar.