Establishment of an apoptosis-resistant and growth-controllable cell line by transfecting with inducible antisense c-Jun gene

• Published in: Biotechnology and bioengineering Vol. 58; no. 1; pp. 65 - 72
• Main Authors: Kim, Yon Hui, Iida, Takehiro, Fujita, Tetsuo, Terada, Satoshi, Kitayama, Atsushi, Ueda, Hiroshi, Prochownik, Edward V., Suzuki, Eiji
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 05.04.1998; Wiley
• Subjects: Animal cell culture; Animal cells; Animals; antisense; apoptosis; Apoptosis - genetics; Biological and medical sciences; Biotechnology; c-jun; cell cycle; Cell Division - drug effects; Dexamethasone - pharmacology; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; F-MEL; Fundamental and applied biological sciences. Psychology; growth deprivation; Growth kinetics; Mathematical models; Methods. Procedures. Technologies; Mice; Oligonucleotides, Antisense - genetics; Proteins; Proto-Oncogene Proteins c-jun - genetics; Transfection; Tumor Cells, Cultured; Erythroleukemia; Induction; Suppression; Rodentia; Gene expression; Cell line establishment; Vertebrata; Mammalia; Cell line; Transfection; Mouse; Cell death; C-Onc gene; Cell cycle; Antisense DNA; Inhibition; Apoptosis
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/(SICI)1097-0290(19980405)58:1<65::AID-BIT7>3.0.CO;2-S

F‐MEL cells were transfected with the c‐jun antisense gene located downstream of a glucocorticoid‐inducible MMTV promoter, and the obtained cells were named c‐jun AS cells. When the c‐jun AS cells were treated with dexamethasone (DEX) in DMEM supplemented with 10% serum, the growth of the cells was completely suppressed for a duration of 16 days with a high cell viability exceeding 86%. The c‐jun expression in the c‐jun AS cells was suppressed moderately in the absence of DEX and strongly in the presence of DEX. The c‐jun AS cells grew well and reached a density of 106 cells/mL without supplementation of any serum components. Viability was greater than 80% after the cells had been cultured for 8 days in the absence of DEX. The c‐jun AS cells stayed at a constant cell density and high viability above 80% for 8 days when they were cultured in the presence of DEX under serum deprivation. In contrast, the wild type F‐MEL cells were unable to grow and died by apoptosis in 3 days under serum deprivation. Internucleosomal cleavage of DNA, a landmark of apoptosis, was clearly detectable. Thus the c‐jun AS cell line that is resistant to apoptosis induced by serum deprivation and can reversibly and viably be growth‐arrested was established. A dual‐signal model was proposed to explain the experimental result, the interlinked regulation of apoptosis, and growth by c‐jun.© 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:65–72, 1998.