Summary
To explore the effect of NF-κB on bcl-x gene transcription in extended drug resistance leukemia cell line HL-60/E6, drug-resistant subline HL-60/E6 was derived by intermittently exposing HL-60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF-κB-RelA in HL-60/E6 cells. FCM analysis and RT-PCR were used to detect the efficiency of liposome-mediated ODN transfection and the change of bcl-XL. mRNA levels after 5 μmol/L phosphorothioate (PS)-derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL-60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL-60/E6 cells, but in the cytoplasm of HL-60 cells, the efficiency of liposomemediated ODN transfection was significantly higher than that of null ODN (P<0. 01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL-60/E6 cells to 5 μmol/L AS-PS-ODN directed to RelA led to a maximal 40 % decline of bcl-xL mRNA levels within 8 h. The inhibition rate of bcl-XL. mRNA was (15±1. 79) %, (28±2. 34) %, (40±3.47) %, (20±1.54) % in 4 h,6h,8h, 15 h, respectively, but it was less than 15 % in control group. It was concluded that NF-κB was involved in regulating bcl-x transcription. It was suggested that NF-κB was an important factor for drug resistance in leukemia cells.
Similar content being viewed by others
References
Chen F, Castranova V, Shi Xet al. New insights into the role of nuclear factor-κB, a ubiquitous transcription factor in the initiation of diseases. Clin Chem, 1999, 45: 7
Reed J C. Double identity for proteins of the Bcl-2 family. Nature, 1997, 387: 773
Lee H H, Dadgostar H, Cheng Qet al. NF-κB-mediated up-regulation of Bcl-x and Bfl-1/A1 is required for CD40 survival signaling in B lymphocytes. Proc Natl Acad Sci USA, 1999, 96: 9136
Kawasaki A M, Casper M D, Freier S Met al. Uniformly modified 2′-deoxy-2′-fluorophosphorothioate oligonucleotides as nuclease-resistant antisense compounds with high affinity and specificity for RNA targets. J Med Chem, 1993, 36: 831
Reuning U, Wilhelm O, Nishiguchi Tet al. Inhibition of NF-κB-RelA expression by antisense oligodeoxynucleotides suppression synthesis of urokinase-type plasminogen activator (uPA) but not its inhibitor PAI-1. Nucleic Acids Res, 1995, 23: 3887
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wenjing, C., Yaozhen, Z., Donghua, Z. et al. Effect of antisense oligodeoxynucleotide directed to NF-κB-RelA on Bcl-xL mRNA in extended drug resistance leukemia cell line HL-60/E6. Current Medical Science 21, 32–34 (2001). https://doi.org/10.1007/BF02888031
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF02888031