Alteration of human breast tumor cell membrane functions by chromosome-mediated gene transfer.
BOT-2 cells (human breast tumor origin) have an impaired ability to utilize exogenous thymidine. Previous studies revealed this deficiency to be the permeation event rather than phosphorylation, since the cells have active thymidine kinase. Chromosome-mediated gene transfer was used to transfer genetic information in the form of metaphase chromosomes, from HeLa-65 cells to the BOT-2 cells, correcting the permease deficiency. Poly-L-ornithine or lipochromes were used for facilitation of chromosome uptake. After selection on HAT medium, transferant clones were isolated at a frequency of 4 x 10(-5) and 1 x 10(-5), respectively. Transferants MGP-1 and MGL-1 are stable after 18 months and have been characterized on the bases of purine and pyrimidine nucleoside uptake, relative thymidine kinase activities, alkaline phosphatase activities, and hydrocortisone-induced alkaline phosphatase activity. MGP-1 demonstrates positive thymidine uptake and incorporates radiolabeled thymidine into DNA. MGL-1 remains thymidine transport-deficient and surveys on HAT by increasing endogenous dihydrofolate reductase activity. Alkaline phosphatase activity in MGL-1 is similar to HeLa-65, 2% of that in BOT-2, and in addition, is inducible 25-30-fold by 3 micro M hydrocortisone. We have separated, genetically, a thymidine permease function from phosphorylation in cells of human origin and have transferred genetic information for the regulation of alkaline phosphatase.
D000469:Alkaline Phosphatase / Q000235:genetics*; D001693:Biological Transport, Active; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism*; D002285:Carcinoma, Intraductal, Noninfiltrating / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002877:Chromosomes, Human; D005260:Female; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics*; D033703:Nucleoside Transport Proteins; D009705:Nucleosides / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism*; D014170:Transformation, Genetic
D026901:Membrane Transport Proteins; D033703:Nucleoside Transport Proteins; D009705:Nucleosides; C025057:thymidine permease; D000469:Alkaline Phosphatase; D013936:Thymidine