沙利度胺诱导急性髓系白血病THP1细胞凋亡和DNA损伤的机制

Mechanism of Thalidomide Induced Apoptosis and DNA Damage in Acute Myeloid Leukemia THP1 Cells

该文研究了沙利度胺对急性髓系白血病THP1细胞凋亡和DNA损伤的诱导作用及分子机制。以体外培养的人急性髓系白血病THP1细胞为研究对象,实验组中加入不同浓度(50、100、150、200、250 μg/mL)的沙利度胺,对照组中加入等体积二甲基亚砜(DMSO)。经处理后,用CCK-8检测沙利度胺对细胞增殖的抑制作用;Calcein/PI法检测药物对细胞凋亡的诱导作用;JC-1荧光染色检测线粒体膜电位的变化;流式细胞仪检测细胞周期;Western blot检测线粒体细胞色素C蛋白、凋亡相关蛋白(cleaved-caspase-3、Bax)和DNA损伤相关蛋白(pATM和γH2AX)的表达水平。结果显示,与对照组相比,实验组沙利度胺明显抑制THP1细胞的生长(P<0.05);沙利度胺对细胞凋亡的诱导作用具有浓度依赖性;沙利度胺导致线粒体膜电位降低(P<0.05),促进线粒体释放细胞色素C蛋白;沙利度胺诱导THP1细胞的G/M期发生阻滞;沙利度胺上调促凋亡相关蛋白(cleaved-caspase-3和Bax)以及DNA损伤应答相关蛋白(pATM和γH2AX)的表达(P<0.05)。这些结果表明,沙利度胺可诱导急性髓系白血病THP1细胞凋亡,其作用机制可能与激活线粒体凋亡途径和诱导DNA损伤有关。

This study investigated the inducing effect and molecular mechanism of thalidomide on apoptosis and DNA damage of acute myeloid leukemia THP1 cells.Taking human acute myeloid leukemia THP1 cells cultured in vitro as the research object,thalidomide with different concentrations (50,100,150,200,250 μg/mL) was added to the experimental group,and an equal volume of DMSO (dimethyl sulfoxide) was added to the control group.After processing,the inhibitory effect of thalidomide on cell proliferation was detected by CCK-8;the inducing effect of drugs on apoptosis was detected by Calcein/PI method;the changes of mitochondrial membrane potential were detected by JC-1 fluorescence staining;the cell cycle was detected by flow cytometry;the level of mitochondrial cytochrome C protein,apoptosis-related proteins (cleaved-caspase-3 and Bax) and DNA damage related proteins (pATM and γH2AX) were detected by Western blot.The results showed that compared with the control group,the thalidomide of experimental group significantly inhibited the growth of THP1 cells (P<0.05);the inducing effect of thalidomide on apoptosis was concentration dependent;thalidomide decreased mitochondrial membrane potential (P<0.05) and promoted the release of cytochrome C protein from mitochondria;thalidomide induced the G/M phase arrest of THP1 cells;thalidomide enhanced the expression of apoptosis-related proteins (cleaved-caspase-3 and Bax) and DNA damage response-related proteins (pATM and γH2AX) (P<0.05).These results suggest that thalidomide can induce apoptosis of acute myeloid leukemia THP1 cells,and its mechanism may be related to activating the mitochondrial apoptosis pathway and inducing DNA damage.