Antisense inhibition of Bax mRNA increases survival of terminally differentiated HL60 cells.
Cell sensitivity to programmed cell death is primarily modulated by members of the Bcl-2 family, as the balance of homodimer or heterodimer formation between proapoptotic and antiapoptotic members defines apoptosis susceptibility in the great majority of cellular contexts. It is, therefore, important to clarify if the Bax protein is limiting for activation of the genetic program of programmed cell death or can be complemented by different Bcl-2 family members, such as Bak or Bad. To gain some insight into the role of Bax in the molecular mechanisms of apoptosis of myeloid cells, we inhibited this gene in all-trans-retinoic acid (ATRA)-treated HL60 cells using the methodology of antisense oligodeoxynucleotides (AS-ODN). Our results indicate that Bax inhibition has no effect on the proliferation and differentiation capacity of HL60 cells. Instead, the survival rate of terminally differentiated Bax-inactivated HL60 (Bax(-) HL60) cells is almost three times higher in respect to control cultures, indicating that in mature granulocytes Bax is not efficiently complemented by others members of the Bcl-2 family proteins.
Manfredini
R
R
Dipartimento di Scienze Biomediche, Sezione di Chimica Biologica, Università di Modena, Italy.
Capobianco
M L
ML
Trevisan
F
F
Rauzi
F
F
Barbieri
D
D
Citro
G
G
Tagliafico
E
E
Ferrari
S
S
D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D002454:Cell Differentiation; D002470:Cell Survival / Q000187:drug effects*; D015195:Drug Design; D018922:HL-60 Cells; D006801:Humans; D009690:Nucleic Acid Conformation; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000737:chemistry; D014212:Tretinoin / Q000494:pharmacology; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; D014212:Tretinoin