[c-myc Antisense Oligodeoxynucleotides Induce Apoptosis of Human Myeloid Leukemia Cell Line HL-60]
In order to study the effects of c-myc antisense phosphorothioate oligodeoxynucleotide in inducing apoptosis of HL-60 cells, the expression of c-myc mRNA was determined by RT-PCR, the morphologic signs of apoptotic cells were observed by transmission electron microscopy, the proportion of apoptotic cells was detected by flow cytometry, and the DNA fragments were analysed by agarose gel electrophoresis. Results of RT-PCR showed marked decrease of c-myc mRNA expression in AspoI and II treated cells. The level of c-Myc protein was decreased 23.8% and 45.4%, respectively, in cells treated with 5 and 10 micro mol/L AspoI, and 38.4% after treatment of 10 micro mol/L AspoII. The apoptotic rates were 23.97% and 52.6% after 10 micro mol/L AspoI treatment and 28.8% and 45.19% after treatment with AspoII 10 micro mol/L for 48 and 72 hours, respectively. while apoptotic cells did not apear in control and sense oligodeoxynucleotide groups. Electron microscopy observation showed the characteritics of apoptosis. A ladder like pattern of DNA fragments was demonstrated on electrophoretogram. The results suggst that c-myc antisense oligodeoxynucleotides could be highly specific gene agents that can suppress the level of c-myc mRNA, decrease the c-Myc proteins and induce apoptosis of HL-60 cells.
Liu
Ting-Bo
TB
The Affiliated Union Hospital, Fujian Medical University, Fujian Institute of Hematology, Fuzhou 350001, China.
Lu
Lian-Huang
LH