标题：亚硒酸钠抑制PI3K/AKT/mTOR信号通路诱导白血病细胞HL-60凋亡的机制研究

      作者：许京淑 1，朱晓健 2，李松 3    (1.恩施土家族苗族自治州中心医院血液病科，湖北 恩施 445000；2.华中科技大学同济医学院附属同济医院血液科，湖北 武汉 430030；3.恩施土家族苗族自治州中心医院血液病实验室 ，湖北 武汉 445000)

      卷次： 2018年29卷3期

      【摘要】 目的 探究亚硒酸钠诱导白血病细胞HL-60凋亡的信号通路及其对信号通路的影响。方法 应用不同浓度的亚硒酸钠处理白血病细胞HL-60，或者应用亚硒酸钠按照不同的时间点处理白血病细胞HL-60，使用流式细胞仪检测亚硒酸钠对HL-60细胞凋亡的影响。应用10 μmol/L亚硒酸钠处理HL-60细胞，Western blot检测p-PI3K、总PI3K、p-AKT、总AKT、p-mTOR和总mTOR的表达情况。在过表达PI3K的HL-60细胞中应用10 μmol/L亚硒酸钠处理白血病细胞HL-60，流式细胞仪检测HL-60细胞的凋亡情况，Western blot检测 p-PI3K、总 PI3K、p-AKT、总AKT、p-mTOR和总mTOR的表达变化。同时以加蒸馏水处理的HL-60细胞为空白对照组。结果 与空白对照组凋亡比例(0.045±0.0013)相比，10 μmol/L及20 μmol/L亚硒酸钠处理组可有效诱导白血病细胞HL-60凋亡[凋亡比例分别为(0.254±0.001 6)和(0.435±0.002 1)]，差异均有统计学意义(P<0.05)。此外与10 μmol/L的亚硒酸钠处理0 h组相比[凋亡比例(0.055±0.001 1)]，10 μmol/L亚硒酸钠作用24 h、48 h、72 h后，HL-60细胞的凋亡显著[凋亡比例分别为(0.179±0.0018)、(0.384±0.0023)和(0.535±0.0034)]，差异均有统计学意义(P<0.05)。亚硒酸钠处理细胞能下调PI3K、AKT、mTOR的磷酸化水平，但对总的PI3K、AKT以及mTOR的表达没有影响。在过表达PI3K的细胞中，AKT、mTOR表达上调，而亚硒酸钠可逆转过表达PI3K导致的PI3K/AKT/mTOR信号通路的过度激活。过表达PI3K使细胞凋亡被抑制，而亚硒酸钠可诱导过表达PI3K的细胞凋亡增加。结论 亚硒酸钠可通过抑制PI3K/AKT/mTOR信号通路的激活，诱导白血病细胞HL-60凋亡。
      【关键词】 亚硒酸钠；人白血病细胞株HL-60；磷脂酰肌醇3-激酶；蛋白激酶 B；哺乳动物雷帕霉素靶蛋白
      【中图分类号】 R733.7 【文献标识码】 A 【文章编号】 1003—6350（2018）03—0300—05

Mechanism of sodium selenite-induced apoptosis of leukemia cell line HL-60 by inhibiting PI3K/AKT/mTORpathway.
XU Jing-shu 1, ZHU Xiao-jian 2, LI Song 3. 1. Department of Hematology, the Central Hospital of Tujia and MiaoAutonomous Prefecture of Enshi, Enshi 445000, Hubei, CHINA; 2. Department of Hematology, Tongji Hospital of TongjiMedical College of Huazhong University of Science & Technology, Wuhan 430030, Hubei, CHINA; 3. HematologyLaboratory, the Central Hospital of Tujia and Miao Autonomous Prefecture of Enshi, Enshi 445000, Hubei, CHINA
【Abstract】 Objective To investigate the signal pathway for sodium selenite-induced apoptosis of leukemia cellline HL-60 and sodium selenite's effects on the signaling pathway. Methods Treating leukemia cell line HL-60 withdifferent concentrations of sodium selenite or using sodium selenite to deal with HL-60 cells at different time points, theeffect of sodium selenite on the apoptosis of HL-60 cells was detected by flow cytometry. HL-60 cells were treated with10 μmol/L sodium selenite, and the expression of p-phosphatidylinositol 3-kinase (PI3K), total (PI3K), p-AKT, totalAKT, p-mammalian target of rapamycin (mTOR), and total (mTOR) were detected by Western blot. The PI3K-overex-pressing HL-60 cells were treated with 10 μmol/L sodium selenite, and flow cytometry was performed to detected theapoptosis of HL-60 cells. Besides, Western blot was performed to detect the expression of p-PI3K, total PI3K, p-AKT,total AKT, p-mTOR, and total mTOR. Results Compared with the blank control group, 10 μmol/L and 20 μmol/L sodi-um selenite can induce the apoptosis of leukemia cells HL-60: the apoptosis ratio (0.045±0.001 3) vs (0.254±0.001 6)and (0.435±0.002 1), P<0.05. In 10 μmol/L sodium selenite treatment condition, comparing with 0 h treatment (the ratioof apoptosis), the treatment of 24 h, 48 h and 72 h significantly induced the apoptosis of HL-60 cells: the apoptosis ratio(0.055±0.001 1) vs (0.179±0.001 8), (0.384±0.002 3), (0.535±0.003 4), P<0.05. Sodium selenite could down-regulate thephosphorylation levels of PI3K, AKT and mTOR, but had no effect on the total expression of PI3K, AKT and mTOR. InPI3K-overexpressing HL-60 cells, the expression of AKT and mTOR were up-regulated, but sodium selenite could inhib-it the over-activation of the PI3K/AKT/mTOR pathway resulting from the overexpression of PI3K. The overexpressionof PI3K could inhibit the apoptosis, but sodium selenite could induce the increase of apoptosis of PI3K-overexpressingcells. Conclusion Sodium selenite can induce the apoptosis of leukemia cell line HL-60 by inhibiting the activation ofPI3K/AKT/mTOR pathway.
      【Key words】 Sodium selenite; Human leukemia cell line HL-60; Phosphatidylinositol 3-kinase (PI3K); Proteinkinase B (PKB); Mammalian target of rapamycin (mTOR)·论 著·doi:10.3969/j.issn.1003-6350.2018.03.002