Establishment of an apoptosis-resistant and growth-controllable cell line by transfecting with inducible antisense c-Jun gene.
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 10(6) 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.
Kim
Y H
YH
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan. sarah@bio.t.u-tokyo.ac.jp
Iida
T
T
Fujita
T
T
Terada
S
S
Kitayama
A
A
Ueda
H
H
Prochownik
E V
EV
Suzuki
E
E
D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D016376:Oligonucleotides, Antisense; D016755:Proto-Oncogene Proteins c-jun; D003907:Dexamethasone