title: Correlation of growth capacity of cells in hard agarose with successful transfection by the activated c-Ha-ras oncogene and in vivo proliferative capacity at metastatic sites.
abstract: The purpose of this study was to determine whether the degree of anchorage-independent growth of rodent or human cells in increasing concentrations of agarose correlated with successful transfection of the cells with an activated c-Ha-ras oncogene and tumorigenicity in nude mice. NIH 3T3 cells, C3H 10T1/2 fibroblasts, four clones of the murine K-1735 melanoma with different metastatic capacities and the TE85 human osteogenic sarcoma line were transfected with plasmids containing the 6.6-kilobase BamHI fragment of the mutant human c-Ha-ras gene and the neo gene, which confers resistance to neomycin (pSV2-neoEJ). Cells transfected with pSV2-neo, a plasmid containing the neo gene, served as controls. Cells from parental or transfected lines (selected by Geneticin) were plated into medium containing 0.3%, 0.6% 0.9%, or 1.2% agarose. These cells were also injected subcutaneously and intravenously into nude mice. The production of tumor cell colonies in dense agarose (greater than or equal to 0.6%) correlated with successful transfection with pSV2-neoEJ and production of experimental metastases in the lung of nude mice. We conclude that the degree of anchorage-independent growth of cells predicts successful transfection with activated c-Ha-ras oncogene and tumorigenic behavior in vivo. Thus this technique may be useful for the detection of cells transfected with transforming oncogenes.
authors.affiliation: Department of Cell Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
authors.forename: I J
authors.initials: IJ
authors.lastname: Fidler
authors.forename: L M
authors.initials: LM
authors.lastname: Li
authors.forename: H N
authors.initials: HN
authors.lastname: Ananthaswamy
authors.forename: N
authors.initials: N
authors.lastname: Esumi
authors.forename: R
authors.initials: R
authors.lastname: Radinsky
authors.forename: J E
authors.initials: JE
authors.lastname: Price
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002455:Cell Division; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D011905:Genes, ras; D006801:Humans; D008546:Melanoma, Experimental / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009362:Neoplasm Metastasis; D012516:Osteosarcoma / Q000473:pathology*; D010957:Plasmids; D012685:Sepharose; D014162:Transfection
chemical_list: D004273:DNA, Neoplasm; D012685:Sepharose