Failure of RB1 to reverse the malignant phenotype of human tumor cell lines.
In addition to retinoblastoma and osteosarcoma, mutation of both alleles of the RB1 gene occurs frequently in several other types of tumors. In order to evaluate the role of RB1 in cancer, the wild type RB1 gene was introduced into the RB1-deleted breast cancer cell line MDA-468-S4 and retinoblastoma cell lines WERI-Rb1 and Y-79. The RB1 complementary DNA was under control of the inducible murine metallothionein promoter in MDA-468-S4 and the thymidine kinase promoter in the retinoblastoma lines. The protein, p110RB1, produced from the exogenously introduced gene appeared normal by immunoprecipitation, Western blot analysis, and nuclear localization and also showed normal cell cycle-dependent phosphorylation and an ability to bind to E1a protein. No changes in growth rate or morphology were observed in either of the reconstituted cell types. Expression of p110RB1 in MDA-468-S4 did not affect anchorage-independent growth when measured by colony formation in soft agar. Although the ability of WERI-Rb1 cells expressing p110RB1 to form colonies in methylcellulose was reduced, the reconstituted retinoblastoma cell lines formed intraocular tumors in immunodeficient mice with the same efficiency as the RB1-negative parent cell lines and the tumors produced by the RB1-reconstituted cells continued to express p110RB1. These experimental results suggest that the malignant phenotype is little affected by the replacement of p110RB1 and that RB1 is a relatively weak tumor suppressor gene.
Muncaster
M M
MM
Department of Molecular and Medical Genetics, University of Toronto, Ontario, Canada.
Cohen
B L
BL
Phillips
R A
RA
Gallie
B L
BL
D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics*; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D002872:Chromosome Deletion; D005134:Eye Neoplasms / Q000235:genetics*; D005260:Female; D016161:Genes, Retinoblastoma; D006367:HeLa Cells; D006801:Humans; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012175:Retinoblastoma / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D011993:Recombinant Fusion Proteins; D008668:Metallothionein; D013937:Thymidine Kinase