本文選題：賴氨酸特異性去甲基化酶 + N-myc下游調(diào)節(jié)基因��； 參考：《山東醫(yī)藥》2017年38期

【摘要】：目的觀察賴氨酸特異性去甲基化酶1(LSD1)、N-myc下游調(diào)節(jié)基因1(NDRG1)基因沉默對(duì)人卵巢癌SKOV3細(xì)胞遷移能力的影響,并探討兩者的作用關(guān)系。方法取人卵巢癌SKOV3細(xì)胞株,通過(guò)shRNA方法建立誘導(dǎo)型干擾LSD1的SKOV3細(xì)胞株(LSD1-shRNA-SKOV3),將其分為對(duì)照組、Dox組、轉(zhuǎn)染組和聯(lián)合組。對(duì)照組用水處理,Dox組用100 ng/m L Dox處理,轉(zhuǎn)染組轉(zhuǎn)染NDRG1 siRNA,聯(lián)合組轉(zhuǎn)染NDRG1 siRNA的同時(shí)加入100 ng/m L Dox處理。采用實(shí)時(shí)熒光定量PCR和Western blotting法分別檢測(cè)4組LSD1、NDRG1基因mRNA和蛋白表達(dá);染色質(zhì)免疫沉淀ChIP法和實(shí)時(shí)熒光定量PCR法分析對(duì)照組和Dox組NDRG1基因啟動(dòng)子區(qū)組蛋白H3第4位賴氨酸的二甲基化(H3K4me2)程度;Transwell小室測(cè)算4組細(xì)胞遷移率。結(jié)果 Dox組、轉(zhuǎn)染組、聯(lián)合組、對(duì)照組LSD1mRNA表達(dá)分別為0.407±0.029、0.936±0.024、0.413±0.018、0.941±0.035,蛋白表達(dá)分別為0.306±0.013、0.879±0.036、0.341±0.057、0.893±0.052,Dox組、聯(lián)合組與對(duì)照組相比,P均0.05。Dox組、轉(zhuǎn)染組、聯(lián)合組、對(duì)照組NDRG1 mRNA表達(dá)分別為0.791±0.045、0.107±0.016、0.165±0.021、0.239±0.027,蛋白表達(dá)分別為0.907±0.005、0.130±0.006、0.216±0.019、0.358±0.062,Dox組、轉(zhuǎn)染組、聯(lián)合組與對(duì)照組相比,聯(lián)合組與Dox組、轉(zhuǎn)染組相比,P均0.05。Dox組、對(duì)照組細(xì)胞NDRG1基因啟動(dòng)子區(qū)H3K4me2水平分別為3.32±0.41、0.83±0.17,兩組相比P0.01。Dox組、轉(zhuǎn)染組、聯(lián)合組、對(duì)照組細(xì)胞遷移率分別為21.75%±1.816%、79.13%±2.561%、40.13%±2.039%、68.91%±3.167%,Dox組、轉(zhuǎn)染組、聯(lián)合組與對(duì)照組相比,聯(lián)合組與Dox組、轉(zhuǎn)染組相比,P均0.05。結(jié)論 LSD1基因沉默人卵巢癌SKOV3細(xì)胞遷移能力降低,NDRG1基因沉默人卵巢癌SKOV3細(xì)胞遷移能力升高;LSD1通過(guò)降低NDRG1基因啟動(dòng)子區(qū)域H3K4me2水平,抑制NDRG1的表達(dá),從而促進(jìn)卵巢癌SKOV3細(xì)胞轉(zhuǎn)移。
[Abstract]:Objective to investigate the effect of Lysine-specific demethylase (LSD1) -N-myc gene silencing on the migration of human ovarian cancer SKOV3 cells. Methods Human ovarian cancer SKOV3 cell line was selected and induced LSD1 interference SKOV3 cell line (LSD1-shRNA-SKOV3) was established by shRNA. The cells were divided into three groups: control group, transfection group and combined group. The control group was treated with 100 ng/m L Dox, the control group was treated with 100 ng/m L Dox, and the combined group was treated with NDRG1 siRNA and 100 ng/m L Dox. Real-time fluorescence quantitative PCR and Western blotting were used to detect the mRNA and protein expression of NDRG1 gene in four groups. Chromatin immunoprecipitation (ChIP) and real-time fluorescence quantitative PCR (PCR) were used to analyze the level of dimethylated H3K4me2 of NDRG1 gene promoter H3 in control group and Dox group. 緇撴灉 Dox緇，

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