本文關(guān)鍵詞： 涎腺腺樣囊性癌 正丁酸鈉 高遷移率蛋白-1 toll樣受體-4 侵襲遷移　出處：《遵義醫(yī)學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：體外觀察不同濃度高遷移率蛋白B1抑制劑—正丁酸鈉對涎腺腺樣囊性癌細(xì)胞株ACC-2和ACC-M侵襲、遷移的影響并初步探討其作用機制。 方法：1.用不同濃度的正丁酸鈉作用ACC-2、ACC-M細(xì)胞24小時，MTT法測定細(xì)胞的增殖率，探尋藥物作用的最佳濃度，采用普通光學(xué)倒置顯微鏡觀察細(xì)胞的形態(tài)變化。2.Transwell小室實驗檢測藥物對ACC-2、ACC-M細(xì)胞侵襲、遷移能力的影響。3.通過Western-blot和實時熒光定量PCR分別檢測藥物作用后兩種細(xì)胞HMGB1和TLR4mRNA和蛋白的表達(dá)。 結(jié)果：1.正丁酸鈉在0.625mM、1.25mM、2.5mM、5mM、10mM濃度時對ACC-2、ACC-M細(xì)胞具有顯著的抑制增殖作用，且抑制作用隨濃度升高而增強（P0.05）。2.Transwell小室實驗證實五個濃度組的正丁酸鈉均可抑制ACC-M細(xì)胞體外侵襲和遷移能力，2.5mM、5mM、10mM濃度組的藥物可以抑制ACC-2細(xì)胞的遷移能力，與對照組相比，差異具有統(tǒng)計學(xué)意義（P0.05）；五個濃度組的藥物對ACC-2細(xì)胞侵襲能力的影響與對照組相比無明顯差異（P0.05）。3.Western-blot和實時熒光定量PCR結(jié)果顯示五個濃度組的正丁酸鈉處理ACC-M細(xì)胞后，可下調(diào)細(xì)胞HMGB1和TLR4蛋白及mRNA的表達(dá)，且具有濃度依賴性，與對照組相比，差異具有統(tǒng)計學(xué)意義（P0.05）；2.5mM、5mM、10mM濃度組的藥物可以下調(diào)ACC-2細(xì)胞HMGB1和TLR4mRNA及蛋白的表達(dá)（P0.05）。相關(guān)性分析顯示：TLR4蛋白表達(dá)的降低與HMGB1的抑制呈正相關(guān)（r=0.810，P0.05）。 結(jié)論：HMGB1的表達(dá)對涎腺腺樣囊性癌細(xì)胞的體外生物學(xué)行為發(fā)揮著重要作用，抑制其表達(dá)可以顯著降低ACC-M細(xì)胞的侵襲和遷移能力，，抑制劑濃度較高時還能降低ACC-2細(xì)胞的遷移能力，TLR4可能作為HMGB1的受體，參與HMGB1對涎腺腺樣囊性癌細(xì)胞體外生物學(xué)行為的調(diào)節(jié)，為篩選臨床靶向藥物治療涎腺腺樣囊性癌提供初步的理論依據(jù)。
[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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R739.87

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