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
Objective To investigate the effects of quercetin on cell morphology, expression of promye-locytic leukemia (PML) mRNA and PML protein localization of NB4, HL-60 cells. Methods Cells mor-phology assayed by Wright’ s ...Objective To investigate the effects of quercetin on cell morphology, expression of promye-locytic leukemia (PML) mRNA and PML protein localization of NB4, HL-60 cells. Methods Cells mor-phology assayed by Wright’ s stain, fluorescence stain, and PML mRNA expression by RT-PCR, PML protein localization by immuno-fluorescence. Results The typical apoptosis was found in NB4 and HL-60 cells after treatment with quercetin. Immuno-fluorescence analysis showed after treatment with quercetin, the fusion pro-tein disappeared in NB4 cells, PML protein relocated, then degraded, and that also seen in HL-60 cells. The expression of PML mRNA is not changed in quercetin-treated cells. Conclusion PML play the role of apop-tosis inducer in leukemia cells at the translational level, quercetin can inhibit the proliferation of leukemia cells, and induce NB4, HL-60 cells apoptosis.展开更多
Arsenic Trioxide (ATO) is widely acknowledged as the treatment of choice for Acute Promyelocytic Leukemia (APL). It is a “two-sided” drug since it can induce differentiation or kill APL and other tumor cells accordi...Arsenic Trioxide (ATO) is widely acknowledged as the treatment of choice for Acute Promyelocytic Leukemia (APL). It is a “two-sided” drug since it can induce differentiation or kill APL and other tumor cells according to the dosage. Part of the cytotoxic effects of ATO on APL cells is due to its pro-oxidant activity, a characteristic which ATO shares with a number of other compounds, including high doses of ascorbate (ASC). In a comparative investigation on the cytotoxic effects of both ATO and ASC on HL60 (APL) cell lines, in Vitro, we have been able to confirm the known cytotoxic effects of ATO, but, more importantly, we have demonstrated that ASC is significantly more effective than ATO, in killing these cancer cells in Vitro, when the concentrations are maintained within the millimolar (mM) range, i.e. the range of plasma concentrations at which ASC induces oxidative damage to tumor cells. Since these plasma levels can be reached only by the intravenous administration of high doses of ASC, we propose that intravenous high doses of ASC may represent a potentially revolutionary new approach in the management of APL.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.
基金Partially supported by the National Natural Science Foundation ofchina (36970328)
文摘Objective To investigate the effects of quercetin on cell morphology, expression of promye-locytic leukemia (PML) mRNA and PML protein localization of NB4, HL-60 cells. Methods Cells mor-phology assayed by Wright’ s stain, fluorescence stain, and PML mRNA expression by RT-PCR, PML protein localization by immuno-fluorescence. Results The typical apoptosis was found in NB4 and HL-60 cells after treatment with quercetin. Immuno-fluorescence analysis showed after treatment with quercetin, the fusion pro-tein disappeared in NB4 cells, PML protein relocated, then degraded, and that also seen in HL-60 cells. The expression of PML mRNA is not changed in quercetin-treated cells. Conclusion PML play the role of apop-tosis inducer in leukemia cells at the translational level, quercetin can inhibit the proliferation of leukemia cells, and induce NB4, HL-60 cells apoptosis.
文摘Arsenic Trioxide (ATO) is widely acknowledged as the treatment of choice for Acute Promyelocytic Leukemia (APL). It is a “two-sided” drug since it can induce differentiation or kill APL and other tumor cells according to the dosage. Part of the cytotoxic effects of ATO on APL cells is due to its pro-oxidant activity, a characteristic which ATO shares with a number of other compounds, including high doses of ascorbate (ASC). In a comparative investigation on the cytotoxic effects of both ATO and ASC on HL60 (APL) cell lines, in Vitro, we have been able to confirm the known cytotoxic effects of ATO, but, more importantly, we have demonstrated that ASC is significantly more effective than ATO, in killing these cancer cells in Vitro, when the concentrations are maintained within the millimolar (mM) range, i.e. the range of plasma concentrations at which ASC induces oxidative damage to tumor cells. Since these plasma levels can be reached only by the intravenous administration of high doses of ASC, we propose that intravenous high doses of ASC may represent a potentially revolutionary new approach in the management of APL.
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。
