Toll样受体9激活对肝癌细胞生长及NF-κB、IRF-7表达的影响

发布时间：2018-04-13 10:33

  本文选题：TLR9 + Hep　； 参考：《宁夏医科大学》2015年硕士论文

【摘要】：目的探讨TLR9激活对Hep G2肝癌细胞增殖、侵袭及NF-κB、IRF-7表达的影响。方法1.将Hep G2细胞分为2组:实验组和阴性对照组。实验组用含不同浓度Cp G-ODN(1、4、16μg·m L-1)的培养液培养Hep G2细胞;阴性对照组用Cp G-ODN(0μg·m L-1)的细胞培养液培养Hep G2细胞。采用实时荧光定量PCR法分别检测实验组和对照组Hep G2细胞中TLR9、NF-κB、IRF-7m RNA表达,收集荧光信号,通过PCR扩增曲线取Ct值,计算得出Hep G2细胞中TLR9和NF-κB、IRF-7m RNA的相对表达量;2.实验组用含不同浓度Cp G-ODN(0.25、1、4、16ug·m L-1)的培养液分别培养Hep G2细胞24、48和72h,采用MTT法检测实验组和对照组Hep G2细胞增殖的变化;3.实验组使用含不同浓度Cp G-ODN(1、4、16ug·m L-1)的培养液培养Hep G2细胞24h,Transwell法检测实验组和对照组Hep G2细胞侵袭能力变化。采用SPSS 17.0统计软件处理实验数据,计量资料以X±s表示,对各组进行单因素方差分析,两样本均数多重比较采用LSD-t检验,以P≤0.05为差异有统计学意义。结果1.实验组与阴性对照组比较,Cp G-ODN(4、16μg·m L-1)作用后的Hep G2细胞中TLR9m RNA、NF-κB m RNA、IRF-7 m RNA的表达增高(P0.05),Cp G-ODN(1μg·m L-1)作用后Hep G2细胞表达的TLR9m RNA、NF-κB m RNA、IRF-7 m RNA与阴性对照组比较差异无统计学意义(P0.05);2.Cp G-ODN(1、4、16μg·m L-1)激动TLR9后可促进Hep G2细胞增殖,且以48h最为显著,并存在剂量依赖关系,Cp G-ODN(0.25μg·m L-1)组与阴性对照组比较差异无统计学意义(P0.05);3.Cp G-ODN(4、16μg·m L-1)激动TLR9可增强Hep G2细胞的侵袭能力(P0.05),Cp G-ODN(1ug·m L-1)组与阴性对照组比较差异无统计学意义(P0.05)。结论1.TLR9的激活可增强Hep G2细胞增殖及侵袭能力2.TLR9可能通过激活NF-k B信号通路促进肝癌细胞生长
[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 卤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，

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