[1]肖蓬莉,郭淑利,王双琳,等.三氧化二砷联合雷帕霉素对急性髓系白血病THP-1细胞增殖、凋亡及自噬的影响[J].新乡医学院学报,2020,37(9):818-823.[doi:10.7683/xxyxyxb.2020.09.003]
　XIAO Pengli,GUO Shuli,WANG Shuanglin,et al.Effects of arsenic trioxide combined with rapamycin on proliferation，apoptosis and autophagy of acute myeloid leukemia THP-1 cells[J].Journal of Xinxiang Medical University,2020,37(9):818-823.[doi:10.7683/xxyxyxb.2020.09.003]
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三氧化二砷联合雷帕霉素对急性髓系白血病THP-1细胞增殖、凋亡及自噬的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年9
页码:
818-823
栏目:
基础研究
出版日期:
2020-09-05

文章信息/Info

Title:
Effects of arsenic trioxide combined with rapamycin on proliferation，apoptosis and autophagy of acute myeloid leukemia THP-1 cells
作者:
肖蓬莉1; 郭淑利1; 王双琳1; 2; 刘思哲1; 2; 彭　靓1; 王万里1; 王松云1; 王慧睿1
（1.郑州大学附属洛阳中心医院血液科，河南　洛阳　471009；2.新乡医学院研究生院，河南　新乡　453003）
Author(s):
XIAO Pengli1; GUO Shuli1; WANG Shuanglin1; 2; LIU Sizhe1; 2; PENG Liang1; WANG Wanli1; WANG Songyun1; WANG Huirui1
(1.Department of Hematology，Luoyang Central Hospital Affiliated to Zhengzhou University，Luoyang 471009，Henan Province，China;2.Graduate School，Xinxiang Medical University，Xinxiang 453003，Henan Province，China)
关键词:
三氧化二砷; 雷帕霉素; THP-1细胞; 增殖; 凋亡; 自噬
Keywords:
arsenic trioxide; rapamycin; THP-1 cell; proliferation; apoptosis; autophagy
分类号:
R714.22
DOI:
10.7683/xxyxyxb.2020.09.003
文献标志码:
A
摘要:
目的　探讨三氧化二砷（As2O3）单用或联合雷帕霉素(Rapa)对急性髓系白血病THP-1细胞增殖、凋亡和自噬的影响及其可能的分子机制。方法　将对数生长期急性髓系白血病THP-1细胞分为对照组、As2O3单药组、Rapa单药组及As2O3+Rapa组。As2O3、Rapa单药组分别应用终浓度0.75、1.50、3.00、6.00、12.00 μmol·L-1 As2O3以及10、20、40、80、160 nmol·L-1Rapa干预细胞，As2O3+Rapa组细胞给予3 mol·L-1 As2O3和 40 mmol·L-1 Rapa干预，对照组细胞采用正常培养基培养。应用细胞计数试剂盒-8（CCK-8）检测3 μmol·L-1As2O3+40 nmol·L-1Rapa联合用药对THP-1细胞的增殖抑制率；流式细胞术检测各组THP-1细胞的凋亡率；实时荧光定量聚合酶链反应检测各组THP-1细胞中自噬相关基因Beclin1、LC3和p62 mRNA的表达；Western blotting 检测各组THP-1细胞中自噬相关蛋白微管相关蛋白1轻链3（LC3）-II/LC3-I的表达。结果　As2O3单独用药和Rapa单独用药时，随着药物浓度增加对急性髓系白血病THP-1细胞的增殖抑制作用逐渐增强，呈剂量依赖关系(P<0.05)。As2O3和Rapa对急性髓系白血病THP-1细胞的IC50分别为5.794 μmol·L-1和77.274 nmol·L-1；As2O3+Rapa组急性髓系白血病THP-1细胞增殖抑制率显著高于As2O3、Rapa单药组(P<0.05)；与As2O3、Rapa单药组比较，As2O3+Rapa组THP-1细胞中自噬相关基因Beclin-1和LC3 mRNA表达水平显著升高(P<0.05)，p62 mRNA 表达水平显著降低(P<0.05)。与As2O3和Rapa单药组比较，As2O3+Rapa组THP-1细胞中LC3-II/LC3-I比值显著增高(P<0.05)。结论　As2O3联合Rapa可协同抑制THP-1细胞增殖，诱导THP-1细胞凋亡，其效应可能通过引起THP-1细胞过度自噬实现。
Abstract:
Objective　To observe the effects of arsenic trioxide (As2O3) combined with rapamycin（Rapa） on the proli-feration，apoptosis and autophagy of acute myeloid leukemia THP-1 cells，and investigate the underlying mechanisms.Methods　The THP-1 cells in the logarithmic growth stage were divided into the control group，the As2O3 treatment group,the Rapa treatment group and the As2O2+Rapa group.The As2O3 treatment group and the Rapa treatment group were treated with 0.75,1.50,3.00,6.00,12.00 μmol·L-1 As2O3 and 10,20,40,80 and 160 nmol·L-1 Rapa，respectively;the cells in As2O2+Rapa grouup were treated with 3 μmol·L-1 As2O2 and 40 nmol·L-1 Rapa;the cells in blank control group were cultured in normal medium.The inhibitory effects of As2O3 and Rapa alone，as well as in a combination of As2O3 and Rapa on THP-1 cell proliferation were detected by using cell counting kit-8 method;the apoptosis of THP-1 cells in the blank control group，As2O3 treatment group，Rapa treatment group and As2O3+Rapa group was detected by using flow cytometry;the expressions of autophagy associated gene Beclin1,microtublule-associated 1 light chain 3（LC3） and p62 mRNA and protein in THP-1 cells in the four groups were measured by using real-time quantitative polymerase chain reaction and Western blotting，respectively.Results　The growth inhibition rate of As2O3 or Rapa alone on the cell proliferation increased with the augment of drug concentration in a concentration-dependent manner(P<0.05).The IC50 of As2O3 and Rapa on THP-1 cells were 5.794 μmol·L-1and 77.274 nmol·L-1，respectively.The cell proliferation inhibition rate and apoptosis ratio in the As2O3+Rapa group were significantly higher than those in the As2O3 treatment group and the Rapa treatment group(P<0.05).The expression of autophagy related genes Beclin-1,LC-3 mRNA in the THP-1 cells in the As2O3+Rapa group were significantly higher than those in the As2O3 treatment group and the Rapa treatment group(P<0.05)，the expression of p62 mRNA was significantly lower than that in the As2O3 treatment group and the Rapa treatment group(P<0.05).The ratio of LC3-II/LC3-I in the As2O3 +Rapa group was significantly higher than that in the As2O3 treatment group and the Rapa treatment group(P<0.05).Conclusion　Combination of As2O3 and Rapa can inhibit THP-1 cell proliferation，induce THP-1 cell apoptosis,which may be achieved by inducing excessive autophagy of THP-1 cells.

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更新日期/Last Update: 2020-09-05