新型反义寡核苷酸F951对HL60细胞Bcl-2基因表达及细胞增殖的抑制作用

Down-regulation of Bcl-2 expression and inhibition of HL60 cells proliferation by F951, a novel Bcl-2 antisense oligodeoxynucleotide

目的 研究新型反义寡核苷酸F951对白血病细胞Bcl- 2基因表达及细胞增殖的影响。方法 不同浓度的F951与HL60细胞共培养后,采用台盼蓝拒染法、MTT比色法检测HL60细胞增殖,用流式细胞术和RT -PCR法检测Bcl- 2蛋白及其mRNA表达,DNA片段化分析法检测凋亡细胞。结果 5, 10, 20μmol·L-1F951分别与HL60细胞共孵育1~5d,细胞生长明显受抑,这一抑制作用随F951浓度的升高和作用时间的延长而增强。MTTA值与未处理组相比F951 5, 10, 20μmol·L-1组细胞抑制率分别为: 20 .56%、37 .66%、54 .11%与对照组相比差异均有显著性;F951处理后HL60细胞Bcl 2mRNA水平下降,Bcl- 2蛋白减少;凝胶电泳可见典型的梯形带,且随着浓度提高诱导DNALadder的作用更加明显。结论 F951可下调Bcl -2基因表达,促进细胞凋亡而抑制白血病细胞增殖。

Aim To investigate whether F951,a novel Bcl-2 antisense oligodeoxynucleotide,down-regulates Bcl-2 expression in HL60 cells and inhibits HL-60 cells proliferation.Methods HL60 cells were cultured with F951 in variant doses.The proliferation of HL60 cells was assayed by MTT and Typan Blue exclusion test.Expression of Bcl-2 protein and its mRNA was measured by FACS and RT-PCR respectively.The apoptotic cells were detected by DNA ladder.Results After HL60 cells were treated with F951 in 5,10,20 μmol·L^(-1) doses respectively for 1～5 days,they showed apparent inhibition of proliferation.With the improvement of the concentration of F951 and the prolongation of the time of treatment, F951 showed stronge effect in the aspect of inhibiting the HL60 cells proliferation.It was determined with MTT method that the inhibition rates of HL60 cells treated with 5,10,20 μmol·L^(-1) F951 were 20.56%, 37.66%, 54.11% respectively. F951 significantly down-regulated the expression of Bcl-2 mRNA and protein in the HL60 cells.Typical DNA ladder was seen from gel electrophoresis. With the improvement of the concentration of F951,it showed more apparent effect in the aspect of inducing DNA ladder.Conclusion F951 can inhibit cells proliferation through down-regulating Bcl-2 gene expression and promoting cells apoptosis in HL60 cells.