[Design of antisense oligodeoxynucleotide targeting at bcl-2 mRNA and observation on its effect on the sensitivity of leukemia cells to arabinosyl cytosine].
The effective target sites for antisense oligodeoxynucleotides (AS-ODN) on bcl-2 mRNA, except its start region, were looked for and their effects on the sensitivity of HL-60 and K562 leukemia cells to arabinosyl cytosine (Ara-C) were observed. The secondary structure of bcl-2 mRNA was simulated with RNAstructure microsoftware, and AS-ODNs, targeting at some sites, were designed and synthesizd with phosphorothioate modifying. The median inhibitory concentration (IC(50)) of Ara-C for HL-60 and K562 cells was determined with MTT method; the apoptosis rate and bcl-2 protein expression level were assayed by flow cytometry. The results showed that 10 micro mol/L bcl-2 AS-ODN combined with Ara-C inhibited the expression of bcl-2 protein, increased apoptosis in HL-60 and K562 cells and decresed IC(50) of Ara-C significantly. The AS-ODN targeting the coding region of bcl-2 mRNA had stronger effect than AS-ODN targeting the translation initiation region. In conclusion, the much more effective sites for antisense oligodeoxynucleotides to target, except translation start region, might provide a new useful way for antisense drug design.
Lei
Xiao-Yong
XY
Institute of Hematology, Medical College of Jinan University, Guangzhou, 510632, China.
Zhang
Yuan
Y
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003561:Cytarabine / Q000494:pharmacology*; D015195:Drug Design; D004357:Drug Synergism; D018922:HL-60 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D020014:K562 Cells; D007938:Leukemia / Q000188:drug therapy / Q000473:pathology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*
D000964:Antimetabolites, Antineoplastic; D016376:Oligonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D003561:Cytarabine