三氧化二砷調控HL-60細胞hTERT基因啟動子及相關轉錄因子NF-κB機制研究
發(fā)布時間:2018-11-18 07:07
【摘要】:目的:研究三氧化二砷(arsenic trioxide,As2O3)對HL-60細胞人端粒酶逆轉錄酶(Human telomerase reverse transcriptase,hTERT)基因啟動子及相關轉錄因子NF-κB的作用機制。方法:1.將不同濃度的As2O3作用HL-60細胞,利用實時熒光定量PCR技術和Western blotting技術分別檢測hTERT、NF-κB在mRNA水平和蛋白水平上的變化2.利用RNAi技術干擾NF-κB基因表達,利用實時熒光定量PCR技術和Western blotting技術,分別檢測HL-60細胞hTERT、NF-κB在mRNA水平和蛋白水平的表達變化,同時利用CCK-8分析法,流式細胞術分別檢測HL-60細胞的增殖和凋亡變化。 結果:1.不同濃度的As2O3作用于HL-60細胞后,hTERT、NF-κB在mRNA和蛋白水平上的表達受到明顯抑制(P0.01)。2.沉默轉錄因子NF-κB的表達,hTERT、NF-κB在mRNA和蛋白水平的表達受到明顯抑制(P0.01)。3.沉默轉錄因子NF-κB的表達,有效的抑制HL-60細胞的增殖(P0.01),以干擾48h最為顯著。4.沉默轉錄因子NF-κB的表達,有效的促進HL-60細胞的凋亡(P0.01)。結論:1.不同濃度的As2O3作用于HL-60細胞后,可促進HL-60細胞凋亡,并呈一定的量效關系。2.沉默轉錄因子NF-κB的表達可促進HL-60細胞的凋亡,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.
【學位授予單位】:蚌埠醫(yī)學院
【學位級別】:碩士
【學位授予年份】:2014
【分類號】:R3411
本文編號: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.
【學位授予單位】:蚌埠醫(yī)學院
【學位級別】:碩士
【學位授予年份】:2014
【分類號】:R3411
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