转铁蛋白受体介导的Bcl-2 shRNA转移对HL-60细胞生长的抑制作用

Inhibition effect of Bcl-2 shRNA on the growth of HL-60 cells via transferring-mediated delivery

目的探讨B cl-2短发夹状RNA(shRNA)表达载体对白血病细胞HL-60生长的抑制作用。方法采用转铁蛋白-多聚乙烯亚胺介导的转染方法将携带绿色荧光蛋白基因的重组B cl-2 shRNA 1、shRNA 2表达载体转入HL-60细胞。通过荧光显微镜和流式细胞仪观察绿色荧光蛋白的表达;采用台盼蓝拒染法计数活细胞;用免疫细胞化学方法检测B cl-2蛋白表达水平。结果B cl-2 shRNA 1、shRNA 2载体分别转入HL-60细胞后,细胞中B cl-2蛋白表达水平均显著降低(P<0.05),转染后48、72、96 h细胞生长明显受到抑制。结论转铁蛋白受体介导的B cl-2 shRNA可显著抑制HL-60细胞生长。

[Objective] To investigate the effect of small hairpin RNA （shRNA） targeting Bcl-2 on the growth of leukemia cell line HL-60. [Methods] Recombinant expression vectors for Bcl-2 shRNAs were transfected into HL-60 via transferrin-polyethylenimine. Transfected cells were visualised using inverted fluorescent microscope and then assayed by flow cytometry. Cell surviving fraction was determined by using the trypan blue dye exclusion assay. The expression levels of Bcl-2 protein were assayed by immunohistochemical method. [Results] The expression levels of Bcl-2 protein from HL-60 cells decreased significantly after Bcl-2 shRNAs treatment （P〈0. 05）. The growth of HL-60 cells was significantly inhibited at 48, 72 and 96 hours after treatment with Bcl-2 shRNA1 or Bcl-2 shRNA2 respectively （P〈0. 05）. [Conclusion] Bcl-2 shRNA expression vectors can effectively inhibit the growth of HL-60 cells.