七叶皂苷钠诱导人单核白血病SHI-1细胞凋亡的研究

Apoptosis of Human Acute Monocytic Leukemic SHI-1 Cells Induced by Sodium Aescinate

目的研究七叶皂苷钠对人急性单核白血病细胞株SHI-1细胞的抑制增殖及诱导凋亡的作用。方法 SHI-1细胞经不同浓度的七叶皂苷钠处理后,MTT法分析七叶皂苷钠对SHI-1细胞增殖的抑制作用,AnnexinⅤ/PI染色流式细胞术(FCM)检测细胞凋亡率,DNA凝胶电泳观察凋亡特异性DNA降解梯形条带,半定量PCR检测凋亡特异性酶Caspase-3 mRNA表达水平。结果MTT法显示七叶皂苷钠能显著抑制SHI-1细胞的增殖,呈时间和剂量依赖性(P<0.05)。经不同浓度的七叶皂苷钠处理24h,AnnexinⅤ/PI染色显示AnnexinⅤ阳性的凋亡细胞率明显上升,DNA琼脂糖凝胶电泳分析出现凋亡特异性的DNA降解梯形条带,半定量PCR显示Caspase-3 mRNA表达水平随药物浓度增加而升高(P<0.05)。结论七叶皂苷钠对SHI-1细胞具有抑制增殖和诱导凋亡的作用,可能是其抗白血病作用途径之一。

Objective To investigate the effects of sodium aescinate on inhibiting proliferation and inducing apoptosis of human acute monocytic leukemic SHI-1 cells. Methods Proliferation of sodium aescinate treated SHI-1 cells was detected by MTT method. The apoptosis rates of SHI-1 cells were analyzed by flow cytometry af-ter annexin V/PI staining. The gel electrophoresis of DNA ladder was used to analyze the bands of DNA degrada-tion, and the expression level of caspase-3 mRNA was tested by reverse transcription-polymerase chain reaction (RT-PCR). Results MTT assay showed that the proliferation of SHI-1 cells was effectively inhibited by sodium aescinate in a time-and-does dependent manner(P<0.05). The Annexinⅴ positive rate of SHI-1 cells after treated with sodium aescinate was significantly increased by FCM analysis(P<0.05). The apoptosis typical DNA fragments of SHI-1 cells treated with sodium aescinate was presented on the gel electrophoresis. The expression level of cas-pase-3 mRNA was up-regulated with increasing concentration of sodium aescinate by RT-PCR detection(P<0.05). Con-clusion Treatment with sodium aescinate can have inhibitory effect on proliferation and induce apoptosis of SHI-1 leukemia cells. Our results provides an experimental evidence for the activity of sodium aescinate on anti-leukemia.