Temperature-sensitive mutants for steroid-sensitive growth of S115 mouse mammary tumor cells

• Published in: Experimental cell research Vol. 186; no. 2; pp. 288 - 298
• Main Authors: Härkönen, Pirkko L., Laaksonen, Esti I.G., Valve, Eeva M., Solic, Nicola, Darbre, Philippa D.
• Format: Journal Article
• Language: English
• Published: Orlando, FL Elsevier Inc 01.02.1990; Elsevier
• Subjects: Androgens - pharmacology; Animals; Biological and medical sciences; Blood; Cell Division - drug effects; Cell physiology; Chloramphenicol O-Acetyltransferase - genetics; Cloning, Molecular; Cytarabine - pharmacology; Ethyl Methanesulfonate; Floxuridine - pharmacology; Fundamental and applied biological sciences. Psychology; Gammaretrovirus - genetics; Gene Expression; Kinetics; Mammary Neoplasms, Experimental - pathology; Mice; Molecular and cellular biology; Mutation; Receptors, Androgen - physiology; RNA, Viral - genetics; Temperature; Testosterone - pharmacology; Transfection; Tumor Cells, Cultured; Steroid; Vertebrata; Mammalia; Cell line; Mouse; Rodentia; Mutation; Cell differentiation; Thermal sensibility
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/0014-4827(90)90308-W

We have generated temperature-sensitive (ts) mutants for steroid-regulated anchorage-independent cell growth. Androgen-responsive S115 + A mouse mammary tumor cells were mutagenized with ethyl methane sulfonate and the variants which were growth-arrested in suspension at the nonpermissive temperature of 41 °C were selected by killing dividing wild-type cells with the DNA synthesis inhibitors 5-fluoro-2′-deoxyuridine or cytosine arabinoside. Fifteen clones were isolated and characterized for morphology and growth properties. Three (ts21, ts27, ts33) of the phenotypic variants were ts for androgen-maintained anchorageindependent growth, two of them (ts27 and ts33) also for growth in monolayer. Growth arrest at 41 °C was not due to a defect in androgen receptor function in any of the mutant cell lines as shown by steroid binding assays and by the androgen-stimulated expression of both endogenous MMTV RNA and the transiently transfected LTR-CAT gene at the nonpermissive temperature. It remains to be determined for clone ts33 whether the defect is in postreceptor events of steroid action or in genes affecting general mechanisms of cell growth. However, since in clones ts21 and ts27 general cell growth remains functional at 41 °C under serum stimulation, defects may be in postreceptor steroid-related pathways. It is hoped that these mutants will provide a useful tool for study of steroid regulation of cell growth and in particular of the property of anchorage-independent growth.