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Caspase-3 antisense oligodeoxynucleotides inhibit apoptosis in γ-irradiated human leukemia HL-60 cells

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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 γ-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 γ-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.

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Acknowledgments

The work was supported by a grant from the National Natural Science Foundation of China (No. 39880008). We are most grateful to Prof. Dwight C. German (Department of psychiatry, University of Texas Southwestern Medical Center) for his suggestions and careful review of the manuscript.

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Authors and Affiliations

  1. 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

    Xiao-Tian Zhang, Tian-Bao Song, Dong-Min Li & Xiao-Ming Li

  2. Department of Histology and Embryology, Guangzhou Medical College, Guangzhou, Guangdong, 510182, China

    Bao-Ling Du

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  1. Xiao-Tian Zhang
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  2. Tian-Bao Song
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  3. Bao-Ling Du
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  4. Dong-Min Li
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Correspondence to Tian-Bao Song.

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The work was supported by a grant from the National Natural Science Foundation of China (No. 39880008).

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Zhang, XT., Song, TB., Du, BL. et al. Caspase-3 antisense oligodeoxynucleotides inhibit apoptosis in γ-irradiated human leukemia HL-60 cells. Apoptosis 12, 743–751 (2007). https://doi.org/10.1007/s10495-006-0018-8

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  • DOI: https://doi.org/10.1007/s10495-006-0018-8

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