title: Caspase-3 antisense oligodeoxynucleotides inhibit apoptosis in gamma-irradiated human leukemia HL-60 cells.
abstract: To study the inhibitory effects of caspase-3 mRNA antisense oligodeoxynucleotides (ASODNs) on apoptosis, we designed four ASODNs targeting different regions of caspase-3 mRNA and transfected them into human leukemia HL-60 cells. The transfected cells were given 10 Gy gamma-irradiation followed by incubation for 18 h and measurement of apoptosis and caspase-3 expression. Our results showed that ASODN-2 targeting the 5' non-coding region of sites -62 to -46, and ASODN-3 targeting the 5' coding region of sites -1 to 16, both reduced apoptosis measured by gel electrophoresis and flow cytometry. Hoechst 33258 staining and TUNEL assay revealed that apoptotic indexes in the ASODN-2 and ASODN-3 groups were significantly lower than those in the untransfected and mismatched oligodeoxynucleotide (MODN) groups. Immunocytochemistry, Western blotting and RT-PCR showed that expression levels of caspase-3 protein and mRNA in both ASODN-2 and ASODN-3 groups were decreased compared with those in the untransfected and MODN groups. In conclusion, caspase-3 mRNA ASODNs can inhibit gamma-radiation-induced apoptosis of HL-60 cells and reduce expression of caspase-3 protein and mRNA. The results suggest that antisense approach may be useful for therapeutic treatment of certain neurodegenerative diseases in which apoptosis is involved.
authors.affiliation: Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, Department of Human Anatomy and Histology-Embryology, School of Medicine, Xi'an Jiaotong University, Xi'an, Shannxi, 710061, China.
authors.forename: Xiao-Tian
authors.initials: XT
authors.lastname: Zhang
authors.forename: Tian-Bao
authors.initials: TB
authors.lastname: Song
authors.forename: Bao-Ling
authors.initials: BL
authors.lastname: Du
authors.forename: Dong-Min
authors.initials: DM
authors.lastname: Li
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D048430:Cell Shape; D053938:DNA Fragmentation; D005720:Gamma Rays; D018922:HL-60 Cells / Q000502:physiology / Q000528:radiation effects; D006801:Humans; D020287:In Situ Nick-End Labeling; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism*
chemical_list: D016376:Oligonucleotides, Antisense; D053148:Caspase 3