Tao Shi, Xu Shan-Qi, Jiao Ming-Xiu, Chen Yu, Fu Cai-Bo, Hu Min
Department of Hematology, The First Affiliated Hospital of Hainan Medical University, Haikou 570100, Hainan Province, China.
Department of Oncology, The Third Affiliated Hospital of Harbin Medical University, Harbin 150000, Heilongjiang Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Oct;28(5):1534-1538. doi: 10.19746/j.cnki.issn.1009-2137.2020.05.018.
To explore the effect of nuclear factor kappa-B (NF-κB) inhibitor pyrrolidine dithiocarbamate (PDTC) on the proliferation and apoptosis of acute leukemia cell HL-60.
HL-60 cells were cultured with PDTC of 0, 25, 50, 100 μmol/L for 24, 48, 72 h. The inhibition rate of cell proliferation was detected by CCK-8 assay. Cell apoptosis was detected by Hoechst staining. Cell cycle was detected by flow cytometry. The expression of B-cell lymphoma-2 (BCL-2), BCL-2 associated X protein (BAX), cyclinD1, activated cysteinyl aspartate specific proteinase (cleaved caspase 3), cleaved caspase 8 and activation of NF-κB signal pathway related protein was detected by Western blot.
After the HL-60 cells were cultured with PDTC of 25, 50, 100 μmol/L for 24, 48, 72 h, the inhibition rate of cell proliferation increased with the enhancement of PDTC concentration at the same time point (r=0.924, P<0.01). At the same PDTC concentration, the inhibition rate of cell proliferation increased with prolonging of time (r=0.952, P<0.01). After HL-60 cell was cultured with PDTC of 25, 50, 100 μmol/L for 48 h, compared with control group, PDTC of 25, 50, 100 μmol/L increased the cell apoptotic rate, arrested cell cycle at G1 phase (P<0.01), the expression of BCL-2, cyclinD1 and p-NF-κB p65 was down-regulated(P<0.05), the expression of BAX, cleaved caspase 3, cleaved caspase 8 was up-regulated(P<0.01). PDTC of 50, 100 μmol/L down-regulated the expression of p-inhibitor of NF-κB (p-IκBα)(P<0.01).
PDTC can inhibit acute leukemia HL-60 cell proliferation and induce cell apoptosis.
探讨核因子κB（NF-κB）抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）对急性白血病细胞HL-60增殖和凋亡的影响。
将HL-60细胞分别用0、25、50、100 μmol/L的PDTC培养24、48、72 h。采用CCK-8法检测细胞增殖抑制率。采用Hoechst染色检测细胞凋亡。采用流式细胞术检测细胞周期。采用蛋白质免疫印迹法检测B细胞淋巴瘤-2（BCL-2）、BCL-2相关X蛋白（BAX）、细胞周期蛋白D1（cyclinD1）、活化的半胱天冬酶3（cleaved caspase 3）、活化的半胱天冬酶8（cleaved caspase 8）及NF-κB信号通路相关蛋白的表达。
HL-60细胞用25、50、100 μmol/L的PDTC培养24、48、72 h后，在同一时间点细胞增殖抑制率随PDTC浓度升高而增加（r=0.924，P<0.01）。在相同PDTC浓度下，细胞增殖抑制率随时间延长而增加（r=0.952，P<0.01）。HL-60细胞用25、50、100 μmol/L的PDTC培养48 h后，与对照组相比，25、50、100 μmol/L的PDTC增加了细胞凋亡率，使细胞周期阻滞于G1期（P<0.01），BCL-2、cyclinD1及p-NF-κB p65的表达下调（P<0.05），BAX、cleaved caspase 3、cleaved caspase 8的表达上调（P<0.01）。50、100 μmol/L的PDTC下调了NF-κB抑制蛋白（p-IκBα）的表达（P<0.01）。
PDTC可抑制急性白血病HL-60细胞增殖并诱导细胞凋亡。
Zotero 插件