Research on 10-23 DNAzymes inhibit the expression of Bcl-2 and induce apoptosis of human hepatoma cells
10.3760/cma.j.issn.1008-6706.2013.21.012
- VernacularTitle:脱氧核酶抑制Bcl-2基因表达诱导人肝癌细胞凋亡的研究
- Author:
Yongguang YANG
;
Mingyi LI
;
Manzhou LIN
;
Guyu ZHANG
- Publication Type:Journal Article
- Keywords:
Deoxyribozyme;
Genes,Bcl-2;
Hepatocellular carcinoma
- From:
Chinese Journal of Primary Medicine and Pharmacy
2013;20(21):3226-3228
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effects of cleavage of Bcl-2 by two DNAzymes on apoptosis of human hepatoma cell line (HepZ1).Methods Two “10-23” DNAzymes(DzT and DzTi) targeting Bcl-2 mRNA and their analogues(DzT' and DzTi') were synthesized and used to cleave Bcl-2 mRNA in vitro and in BEL-7402 cells.The RT-PCR was performed to assess the cleaving efficiency.Expression of Bcl-2 protein was determined by immunofluorescent method.Cell apoptosis was detected by flow cytometry.Results The unmodified Enzymes DzT,and its modified form DzTi,which had an added 3'-inverted thymidine,could effectively cleave Bcl-2 mRNA in vitro.After transfected into BEL-7402 cells,DzTi exhibited more powerful cleaving ability than DzT,significantly down-regulated the level of Bcl-2 protein(P ＜0.01) and inhibited the cell growth(P ＜0.05).The results of flow cytometry suggested that the apoptosis rate of DzT and DzTi significantly increased,appeared apoptotic peak.Cell cycle was delayed in DzT and DzTi group,proportion of cells in G0/G1 increased,S phase cells decreased.Conclusion The synthesized DNAzymes could effectively cleave Bcl-2 mRNA,decrease the level of Bcl-2 protein and induce hepatoma cells apoptosis.
