HMGB1抑制剂对人涎腺腺样囊性癌细胞株侵袭、迁移的影响及机制研究

发布时间：2018-02-25 10:12

本文关键词： 涎腺腺样囊性癌正丁酸钠高迁移率蛋白-1 toll样受体-4 侵袭迁移　出处：《遵义医学院》2014年硕士论文　论文类型：学位论文

【摘要】：目的：体外观察不同浓度高迁移率蛋白B1抑制剂—正丁酸钠对涎腺腺样囊性癌细胞株ACC-2和ACC-M侵袭、迁移的影响并初步探讨其作用机制。方法：1.用不同浓度的正丁酸钠作用ACC-2、ACC-M细胞24小时，MTT法测定细胞的增殖率，探寻药物作用的最佳浓度，采用普通光学倒置显微镜观察细胞的形态变化。2.Transwell小室实验检测药物对ACC-2、ACC-M细胞侵袭、迁移能力的影响。3.通过Western-blot和实时荧光定量PCR分别检测药物作用后两种细胞HMGB1和TLR4mRNA和蛋白的表达。结果：1.正丁酸钠在0.625mM、1.25mM、2.5mM、5mM、10mM浓度时对ACC-2、ACC-M细胞具有显著的抑制增殖作用，且抑制作用随浓度升高而增强（P0.05）。2.Transwell小室实验证实五个浓度组的正丁酸钠均可抑制ACC-M细胞体外侵袭和迁移能力，2.5mM、5mM、10mM浓度组的药物可以抑制ACC-2细胞的迁移能力，与对照组相比，差异具有统计学意义（P0.05）；五个浓度组的药物对ACC-2细胞侵袭能力的影响与对照组相比无明显差异（P0.05）。3.Western-blot和实时荧光定量PCR结果显示五个浓度组的正丁酸钠处理ACC-M细胞后，可下调细胞HMGB1和TLR4蛋白及mRNA的表达，且具有浓度依赖性，与对照组相比，差异具有统计学意义（P0.05）；2.5mM、5mM、10mM浓度组的药物可以下调ACC-2细胞HMGB1和TLR4mRNA及蛋白的表达（P0.05）。相关性分析显示：TLR4蛋白表达的降低与HMGB1的抑制呈正相关（r=0.810，P0.05）。结论：HMGB1的表达对涎腺腺样囊性癌细胞的体外生物学行为发挥着重要作用，抑制其表达可以显著降低ACC-M细胞的侵袭和迁移能力，，抑制剂浓度较高时还能降低ACC-2细胞的迁移能力，TLR4可能作为HMGB1的受体，参与HMGB1对涎腺腺样囊性癌细胞体外生物学行为的调节，为筛选临床靶向药物治疗涎腺腺样囊性癌提供初步的理论依据。
[Abstract]:Aim: to investigate the effect of high mobility protein B1 inhibitor sodium butyrate on the invasion and migration of salivary adenoid cystic carcinoma cell line ACC-2 and ACC-M in vitro. Methods the proliferation rate of ACC-2ACC-M cells was determined by MTT assay with different concentrations of sodium butyrate for 24 hours. The morphological changes of ACC-2ACC-M cells were observed by ordinary optical inverted microscope. 2. Transwell chamber assay was used to detect the invasion of ACC-2ACC-M cells. Western-blot and real-time fluorescence quantitative PCR were used to detect the expression of HMGB1, TLR4mRNA and protein in the two kinds of cells after drug treatment. Results: 1. Sodium butyrate could significantly inhibit the proliferation of ACC-2ACC-M cells at the concentration of 0.625mMU 1.25mMU 2.5mMU 5mMU 10mM, and the effect of sodium butyrate on the proliferation of ACC-2ACC-M cells was significant. The inhibitory effect increased with the increase of concentration. 2. Transwell chamber experiments showed that sodium butyrate could inhibit the invasion and migration of ACC-M cells in vitro. There was no significant difference in the invasion ability of ACC-2 cells between the five concentration groups and the control group. The results of Western-blot and real-time fluorescence quantitative PCR showed that the five concentration groups of sodium butyrate treated ACC-M cells. The expression of HMGB1 and TLR4 protein and mRNA were down-regulated in a concentration-dependent manner. The difference was statistically significant (P 0.05). The drug in the concentration group of 2.5mM and 5mM could down-regulate the expression of HMGB1, TLR4mRNA and protein in ACC-2 cells. The correlation analysis showed that the decrease of the protein expression of TLR4 was positively correlated with the inhibition of HMGB1. Conclusion the expression of HMGB1 plays an important role in the biological behavior of salivary adenoid cystic carcinoma cells in vitro. Inhibition of the expression of the tumor cells can significantly reduce the invasion and migration of ACC-M cells. TLR4 may act as a receptor of HMGB1 and may be involved in the regulation of the biological behavior of salivary adenoid cystic carcinoma cells in vitro by HMGB1 when the concentration of TLR4 is high. To screen clinical targeted drugs for salivary gland adenoid cystic carcinoma to provide a preliminary theoretical basis.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.87

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