本文選題：TLR9 + Hep��； 參考：《寧夏醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的探討TLR9激活對Hep G2肝癌細(xì)胞增殖、侵襲及NF-κB、IRF-7表達的影響。方法1.將Hep G2細(xì)胞分為2組:實驗組和陰性對照組。實驗組用含不同濃度Cp G-ODN(1、4、16μg·m L-1)的培養(yǎng)液培養(yǎng)Hep G2細(xì)胞;陰性對照組用Cp G-ODN(0μg·m L-1)的細(xì)胞培養(yǎng)液培養(yǎng)Hep G2細(xì)胞。采用實時熒光定量PCR法分別檢測實驗組和對照組Hep G2細(xì)胞中TLR9、NF-κB、IRF-7m RNA表達,收集熒光信號,通過PCR擴增曲線取Ct值,計算得出Hep G2細(xì)胞中TLR9和NF-κB、IRF-7m RNA的相對表達量;2.實驗組用含不同濃度Cp G-ODN(0.25、1、4、16ug·m L-1)的培養(yǎng)液分別培養(yǎng)Hep G2細(xì)胞24、48和72h,采用MTT法檢測實驗組和對照組Hep G2細(xì)胞增殖的變化;3.實驗組使用含不同濃度Cp G-ODN(1、4、16ug·m L-1)的培養(yǎng)液培養(yǎng)Hep G2細(xì)胞24h,Transwell法檢測實驗組和對照組Hep G2細(xì)胞侵襲能力變化。采用SPSS 17.0統(tǒng)計軟件處理實驗數(shù)據(jù),計量資料以X±s表示,對各組進行單因素方差分析,兩樣本均數(shù)多重比較采用LSD-t檢驗,以P≤0.05為差異有統(tǒng)計學(xué)意義。結(jié)果1.實驗組與陰性對照組比較,Cp G-ODN(4、16μg·m L-1)作用后的Hep G2細(xì)胞中TLR9m RNA、NF-κB m RNA、IRF-7 m RNA的表達增高(P0.05),Cp G-ODN(1μg·m L-1)作用后Hep G2細(xì)胞表達的TLR9m RNA、NF-κB m RNA、IRF-7 m RNA與陰性對照組比較差異無統(tǒng)計學(xué)意義(P0.05);2.Cp G-ODN(1、4、16μg·m L-1)激動TLR9后可促進Hep G2細(xì)胞增殖,且以48h最為顯著,并存在劑量依賴關(guān)系,Cp G-ODN(0.25μg·m L-1)組與陰性對照組比較差異無統(tǒng)計學(xué)意義(P0.05);3.Cp G-ODN(4、16μg·m L-1)激動TLR9可增強Hep G2細(xì)胞的侵襲能力(P0.05),Cp G-ODN(1ug·m L-1)組與陰性對照組比較差異無統(tǒng)計學(xué)意義(P0.05)。結(jié)論1.TLR9的激活可增強Hep G2細(xì)胞增殖及侵襲能力2.TLR9可能通過激活NF-k B信號通路促進肝癌細(xì)胞生長
[Abstract]:Objective to investigate the effects of TLR9 activation on the proliferation, invasion and expression of NF- 魏 B TLR9-7 in Hep G2 hepatoma cells.Method 1.Hep G2 cells were divided into two groups: experimental group and negative control group.Hep G2 cells were cultured in the culture medium containing different concentrations of CpG-ODN1 (16 渭 g mL -1) in the experimental group, and Hep G2 cells in the negative control group were cultured in the medium containing CP G-ODN(0 渭 g ml -1).Real-time fluorescence quantitative PCR was used to detect the expression of TLR9NF-kB PCR in Hep G2 cells. The fluorescence signals were collected and the Ct values were obtained by PCR amplification curve. The relative expression levels of TLR9 and NF- 魏 BNIRF-7m RNA in Hep G2 cells were calculated and the relative expression levels of TLR9 and NF- 魏 BnirF-7m RNA were calculated.In the experimental group, Hep G2 cells were cultured for 2448 h and 72 h respectively in the medium containing different concentrations of CP G-ODN 0.25g / L and 416ug ml / L, respectively. The proliferation of Hep G2 cells in the experimental group and the control group was detected by MTT assay.The invasiveness of Hep G2 cells in the experimental group and the control group were detected by 24 h transwell method in the culture medium containing different concentrations of CpG-ODN1 (4ug mL -1). The invasion ability of Hep G2 cells in the experimental group and the control group was measured.SPSS 17.0 statistical software was used to process the experimental data. The measurement data were expressed as X 鹵s. The single factor ANOVA was used in each group. The LSD-t test was used to compare the mean of the two samples, and the difference was statistically significant (P 鈮，

本文編號：1744117