【摘要】：目的：研究三氧化二砷(arsenic trioxide，As2O3)對(duì)HL-60細(xì)胞人端粒酶逆轉(zhuǎn)錄酶（Human telomerase reverse transcriptase,hTERT）基因啟動(dòng)子及相關(guān)轉(zhuǎn)錄因子NF-κB的作用機(jī)制。方法：1.將不同濃度的As2O3作用HL-60細(xì)胞，利用實(shí)時(shí)熒光定量PCR技術(shù)和Western blotting技術(shù)分別檢測(cè)hTERT、NF-κB在mRNA水平和蛋白水平上的變化2.利用RNAi技術(shù)干擾NF-κB基因表達(dá)，利用實(shí)時(shí)熒光定量PCR技術(shù)和Western blotting技術(shù)，分別檢測(cè)HL-60細(xì)胞hTERT、NF-κB在mRNA水平和蛋白水平的表達(dá)變化，同時(shí)利用CCK-8分析法，流式細(xì)胞術(shù)分別檢測(cè)HL-60細(xì)胞的增殖和凋亡變化。 結(jié)果：1.不同濃度的As2O3作用于HL-60細(xì)胞后，hTERT、NF-κB在mRNA和蛋白水平上的表達(dá)受到明顯抑制(P0.01)。2.沉默轉(zhuǎn)錄因子NF-κB的表達(dá)，hTERT、NF-κB在mRNA和蛋白水平的表達(dá)受到明顯抑制(P0.01)。3.沉默轉(zhuǎn)錄因子NF-κB的表達(dá)，有效的抑制HL-60細(xì)胞的增殖(P0.01)，以干擾48h最為顯著。4.沉默轉(zhuǎn)錄因子NF-κB的表達(dá)，有效的促進(jìn)HL-60細(xì)胞的凋亡(P0.01)。結(jié)論：1.不同濃度的As2O3作用于HL-60細(xì)胞后,可促進(jìn)HL-60細(xì)胞凋亡，并呈一定的量效關(guān)系。2.沉默轉(zhuǎn)錄因子NF-κB的表達(dá)可促進(jìn)HL-60細(xì)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(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R3411

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本文編號(hào)：2339254