The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a ch...The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.展开更多
目的:探究苦参碱联合化疗药物对人急性髓系白血病HL-60细胞增殖及侵袭力的影响。方法:体外培养人急性髓系白血病HL-60细胞,苦参碱单药组以1.0 g/L苦参碱干预,三氧化二砷单药组以3μmol/L三氧化二砷处理,联合组以终浓度为1.0 g/L的苦参...目的:探究苦参碱联合化疗药物对人急性髓系白血病HL-60细胞增殖及侵袭力的影响。方法:体外培养人急性髓系白血病HL-60细胞,苦参碱单药组以1.0 g/L苦参碱干预,三氧化二砷单药组以3μmol/L三氧化二砷处理,联合组以终浓度为1.0 g/L的苦参碱+3μmol/L三氧化二砷处理,空白组以等量生理盐水处理。MTT法检测HL-60细胞增殖情况,ranswell小室实验检测HL-60侵袭力,蛋白免疫印迹法检测HL-60细胞MMPs蛋白表达情况。结果:苦参碱单药组、三氧化二砷单药组、联合组24 h PIR分别为(18.54±2.54)%,(29.54±2.62)%,(41.45±3.72)%,48 h PIR分别为(27.46±2.55)%,(38.93±3.76)%,(45.64±3.54)%,3组内HL-60的PIR均随着培养时间的增加而增加,其中联合组24 h、48 h PIR均高于同时间点的苦参碱单药组、三氧化二砷单药组,差异有统计学意义(P<0.05)。空白组、苦参碱单药组、三氧化二砷单药组、联合组侵袭细胞分别为(127.64±23.93)个、(94.24±15.23)个、(81.23±12.21)个、(75.13±10.69)个,相对侵袭指数分别为(100.00±0.00)%、(73.44±0.34)%、(62.47±0.15)%、(58.54±0.36)%;MMP-2蛋白相对表达量分别为(0.91±0.05)、(0.39±0.04)、(0.23±0.04)、(0.14±0.03);MMP-9蛋白相对表达量分别为(0.62±0.05)、(0.31±0.05)、(0.27±0.04)、(0.16±0.02)。与空白组比较,苦参碱单药组、三氧化二砷单药组、联合组侵袭细胞数量显著减少,相对侵袭指数及细胞中MMP-2、MMP-9相对表达量降低,且联合组均低于苦参碱单药组、三氧化二砷单药组,差异有统计学意义(P<0.05)。结论:苦参碱联合三氧化二砷通过下调MMP-2、MMP-9蛋白表达有效抑制人急性髓系白血病HL-60细胞增殖及侵袭。展开更多
文摘The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.
文摘目的:探究苦参碱联合化疗药物对人急性髓系白血病HL-60细胞增殖及侵袭力的影响。方法:体外培养人急性髓系白血病HL-60细胞,苦参碱单药组以1.0 g/L苦参碱干预,三氧化二砷单药组以3μmol/L三氧化二砷处理,联合组以终浓度为1.0 g/L的苦参碱+3μmol/L三氧化二砷处理,空白组以等量生理盐水处理。MTT法检测HL-60细胞增殖情况,ranswell小室实验检测HL-60侵袭力,蛋白免疫印迹法检测HL-60细胞MMPs蛋白表达情况。结果:苦参碱单药组、三氧化二砷单药组、联合组24 h PIR分别为(18.54±2.54)%,(29.54±2.62)%,(41.45±3.72)%,48 h PIR分别为(27.46±2.55)%,(38.93±3.76)%,(45.64±3.54)%,3组内HL-60的PIR均随着培养时间的增加而增加,其中联合组24 h、48 h PIR均高于同时间点的苦参碱单药组、三氧化二砷单药组,差异有统计学意义(P<0.05)。空白组、苦参碱单药组、三氧化二砷单药组、联合组侵袭细胞分别为(127.64±23.93)个、(94.24±15.23)个、(81.23±12.21)个、(75.13±10.69)个,相对侵袭指数分别为(100.00±0.00)%、(73.44±0.34)%、(62.47±0.15)%、(58.54±0.36)%;MMP-2蛋白相对表达量分别为(0.91±0.05)、(0.39±0.04)、(0.23±0.04)、(0.14±0.03);MMP-9蛋白相对表达量分别为(0.62±0.05)、(0.31±0.05)、(0.27±0.04)、(0.16±0.02)。与空白组比较,苦参碱单药组、三氧化二砷单药组、联合组侵袭细胞数量显著减少,相对侵袭指数及细胞中MMP-2、MMP-9相对表达量降低,且联合组均低于苦参碱单药组、三氧化二砷单药组,差异有统计学意义(P<0.05)。结论:苦参碱联合三氧化二砷通过下调MMP-2、MMP-9蛋白表达有效抑制人急性髓系白血病HL-60细胞增殖及侵袭。
