title: [Chimeric Ad5F35 adenoviral vector-mediated expression of mutant IκBα induces apoptosis of leukemia cells].
abstract: Constitutive activation of nuclear transcription factor-κB (NF-κB) exists in a variety of leukemia, and induction of apoptosis through blocking NF-κB activation may be an alternative strategy for leukemia treatment. The aim of this study was to investigate the inducing effect of modified adenovirus 5-based adenovirus vector (i.e. chimeric Ad5F35 Vec)-mediated expression of mutant IκBα (IκBαDN) on apoptosis of HL-60 cells. The recombinant Ad5F35-IκBαDN Vec carrying IκBαDN cDNA which deleted the first 1-70 amino acids coding sequences at 5' terminal of human IκBα was transfected into HL-60 cells. The apoptosis, NF-κB DNA binding activity, the expressions of IκBα, cIAP-2 and xIAP in HL-60 cells were detected by DNA binding assay, flow cytometry, real-time quantitative polymerase chain reaction and Western blot respectively. The results showed that apoptosis rates were 22.53 ± 2.999%, 6.08 ± 2.464% and 4.86 ± 1.366% for Ad5F35-IκBαDN Vec-infected or blank vector of Ad5F35-EGFP Vec-transfected and untransfected HL-60 cells respectively, which showed a significant difference between Ad5F35-IκBαDN Vec-transfected and untransfected cells (p < 0.001) and between Ad5F35-IκBαDN Vec-transfected and Ad5F35-EGFP Vec-transfected cells (p < 0.001, p < 0.002), while NF-κB DNA binding activity was decreased, the truncated IκBα was expressed, and IκBα mRNA expression was up-regulated, but the expression of cIAP-2 and xIAP mRNA was down-regulated after transduction for 48 hours. It is concluded that the chimeric Ad5F35 Vec can effectively mediate the expression of IκBαDN cDNA in HL-60 cells, leading to the inhibition of NF-κB DNA binding activity and inducing apoptosis of HL-60 cells.
authors.affiliation: Department of Hematology, Central South University, Changsha, Hunan Province, China. guangpingw@yahoo.com
authors.forename: Guang-Ping
authors.initials: GP
authors.lastname: Wang
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.forename: Hong-Ya
authors.initials: HY
authors.lastname: Xin
authors.forename: Zhao-Jun
authors.initials: ZJ
authors.lastname: Duan
authors.forename: Zhao-Zheng
authors.initials: ZZ
authors.lastname: Jing
authors.forename: San-Qing
authors.initials: SQ
authors.lastname: Tan
authors.forename: Zhen-Hua
authors.initials: ZH
authors.lastname: Qi
authors.forename: Fang-Ping
authors.initials: FP
authors.lastname: Chen
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis; D005822:Genetic Vectors; D018922:HL-60 Cells; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000235:genetics*; D014162:Transfection
chemical_list: D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D000072000:NF-KappaB Inhibitor alpha