三氧化二砷调控HL-60细胞hTERT基因启动子及相关转录因子NF-κB机制研究
发布时间：2018-11-18 07:07
【摘要】：目的：研究三氧化二砷(arsenic trioxide，As2O3)对HL-60细胞人端粒酶逆转录酶（Human telomerase reverse transcriptase,hTERT）基因启动子及相关转录因子NF-κB的作用机制。方法：1.将不同浓度的As2O3作用HL-60细胞，利用实时荧光定量PCR技术和Western blotting技术分别检测hTERT、NF-κB在mRNA水平和蛋白水平上的变化2.利用RNAi技术干扰NF-κB基因表达，利用实时荧光定量PCR技术和Western blotting技术，分别检测HL-60细胞hTERT、NF-κB在mRNA水平和蛋白水平的表达变化，同时利用CCK-8分析法，流式细胞术分别检测HL-60细胞的增殖和凋亡变化。 结果：1.不同浓度的As2O3作用于HL-60细胞后，hTERT、NF-κB在mRNA和蛋白水平上的表达受到明显抑制(P0.01)。2.沉默转录因子NF-κB的表达，hTERT、NF-κB在mRNA和蛋白水平的表达受到明显抑制(P0.01)。3.沉默转录因子NF-κB的表达，有效的抑制HL-60细胞的增殖(P0.01)，以干扰48h最为显著。4.沉默转录因子NF-κB的表达，有效的促进HL-60细胞的凋亡(P0.01)。结论：1.不同浓度的As2O3作用于HL-60细胞后,可促进HL-60细胞凋亡，并呈一定的量效关系。2.沉默转录因子NF-κB的表达可促进HL-60细胞的凋亡，抑制其增殖。
[Abstract]:Aim: to investigate the mechanism of arsenic trioxide (arsenic trioxide,As2O3) on the promoter of human telomerase reverse transcriptase (Human telomerase reverse transcriptase,hTERT) gene and the related transcription factor NF- 魏 B in HL-60 cells. Method 1: 1. HL-60 cells were treated with different concentrations of As2O3. The changes of hTERT,NF- 魏 B at mRNA level and protein level were detected by real-time fluorescence quantitative PCR and Western blotting techniques respectively. RNAi was used to interfere the expression of NF- 魏 B gene, real-time quantitative PCR and Western blotting techniques were used to detect the expression of hTERT,NF- 魏 B in HL-60 cells at the level of mRNA and protein, respectively. Meanwhile, CCK-8 analysis was used to detect the expression of hTERT,NF- 魏 B in HL-60 cells. The proliferation and apoptosis of HL-60 cells were detected by flow cytometry. Results: 1. The expression of hTERT,NF- 魏 B at the level of mRNA and protein was significantly inhibited in HL-60 cells treated with different concentrations of As2O3 (P0.01). Silencing the expression of NF- 魏 B and the expression of hTERT,NF- 魏 B at the level of mRNA and protein were significantly inhibited (P0.01). Silencing the expression of NF- 魏 B effectively inhibited the proliferation of HL-60 cells (P0.01), especially for 48 h. Silencing the expression of NF- 魏 B can effectively promote the apoptosis of HL-60 cells (P0.01). Conclusion 1. Different concentrations of As2O3 could promote the apoptosis of HL-60 cells in a dose-dependent manner. 2. Silencing the expression of NF- 魏 B can promote the apoptosis and inhibit the proliferation of HL-60 cells.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R3411
本文编号：2339254
[Abstract]:Aim: to investigate the mechanism of arsenic trioxide (arsenic trioxide,As2O3) on the promoter of human telomerase reverse transcriptase (Human telomerase reverse transcriptase,hTERT) gene and the related transcription factor NF- 魏 B in HL-60 cells. Method 1: 1. HL-60 cells were treated with different concentrations of As2O3. The changes of hTERT,NF- 魏 B at mRNA level and protein level were detected by real-time fluorescence quantitative PCR and Western blotting techniques respectively. RNAi was used to interfere the expression of NF- 魏 B gene, real-time quantitative PCR and Western blotting techniques were used to detect the expression of hTERT,NF- 魏 B in HL-60 cells at the level of mRNA and protein, respectively. Meanwhile, CCK-8 analysis was used to detect the expression of hTERT,NF- 魏 B in HL-60 cells. The proliferation and apoptosis of HL-60 cells were detected by flow cytometry. Results: 1. The expression of hTERT,NF- 魏 B at the level of mRNA and protein was significantly inhibited in HL-60 cells treated with different concentrations of As2O3 (P0.01). Silencing the expression of NF- 魏 B and the expression of hTERT,NF- 魏 B at the level of mRNA and protein were significantly inhibited (P0.01). Silencing the expression of NF- 魏 B effectively inhibited the proliferation of HL-60 cells (P0.01), especially for 48 h. Silencing the expression of NF- 魏 B can effectively promote the apoptosis of HL-60 cells (P0.01). Conclusion 1. Different concentrations of As2O3 could promote the apoptosis of HL-60 cells in a dose-dependent manner. 2. Silencing the expression of NF- 魏 B can promote the apoptosis and inhibit the proliferation of HL-60 cells.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R3411
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