bcl-2反义核酸提高白血病HL60细胞对米托蒽醌的敏感性

Bcl - 2 antisense oligodeoxynucleotide increases the sensitivity of HL60 cells to mitoxantrone

目的 探讨下调HL6 0细胞bcl 2基因表达对米托蒽醌 (MIT)细胞毒作用的影响。方法 将针对bcl 2基因编码区 141 147密码子的反义寡核苷酸 (ASODN)作用于HL6 0细胞 ,测定其mRNA表达 ,细胞生长 ,细胞凋亡。结果 12 5μmol/LASODN作用于HL6 0细胞 48h可以下调bcl 2mRNA表达 40 %。 0 5 μmol/LMIT作用于经ASODN预处理 48h的HL6 0细胞 5h ,5 0 9%的细胞发生凋亡 ,明显高于MIT单独作用诱导的细胞凋亡 (2 5 6 % ) ;当ASODN与 0 7nmol/LMIT共同作用时 ,细胞存活率 (4 8h)由MIT单独作用时的 73 7%降至 2 5 6 %。结论 bcl 2ASODN可下调bcl 2基因表达 ,促进细胞凋亡而增加MIT抗肿瘤作用。

Objective To explore the effect of antisense oligodeoxynucleotide on down regulating bcl 2 gene expression and increasing the sensitivity of mitroxantrone (MIT) in human myeloid leukemia. Methods Antisense oligodexyonucleotide (ASODN) targeting 141 147 codons of bcl 2 mRNA was applied in HL60 cells. The cell growth, apoptosis and gene expression were measured by trypan blue exclusion, flow cytometry and RT PCR respectively. Results The level of bcl 2 mRNA was decreased by 40% in HL60 cells treated with 12.5 μmol/L ASODN for 48 hours. Mitoxantrone at the concentration of 0.5 μmol/L for 5 hours induced a higher percentage of apoptosis (50.9%) in HL60 cells pretreated with 12.5 μmol/L ASODN for 48 hours than that in control cells (25.6%). Compared with mitoxantrone alone for 48 hours, combination of 12.5 μmol/L ASODN and mitoxantrone reduced the rate of cell survival from 73.7% to 25.6%. Conclusion bcl 2 ASODN can inhibit bcl 2 gene expression, and enhance the cytotoxicity through promoting mitoxantrone induced apoptosis in HL60 cells and hence increase the anti tumor effect of MIT.$$$$