【摘要】：背景和目的 宮頸癌是女性生殖系統(tǒng)中病死率較高的惡性腫瘤之一,其發(fā)病率呈緩慢上升的趨勢。本研究欲通過選用不同濃度蜂膠提取物處理宮頸癌細胞Hela,探討蜂膠對其影響,為臨床治療提供理論指導。 方法 1、采用IHC法檢測蜂膠對Hela中細胞因子分泌的影響； 2、從DNA轉錄調控,RNA和蛋白水平系統(tǒng)研究蜂膠對Hela細胞中炎癥因子的影響； 3、從RNA和蛋白水平研究蜂膠對Hela細胞分化增殖相關基因Egr1、p21和Ki67表達的影響； 結果 1.濃度4、8、16、25μg/mL的蜂膠可以顯著影響Hela細胞中IL-8、TNF-α和IL-10等細胞因子的分泌,同時發(fā)現濃度達到16μg/mL以上時,對巨噬細胞刺激分泌NO可達對照組的7倍以上。 2.濃度4、8、16、25μg/mL的蜂膠能從DNA轉錄調控,RNA和蛋白水平上調節(jié)Hela細胞中炎癥因子iNOS、PTGS-2和IL-8的表達,初步顯示通過NF-κB細胞信號通路起作用。 3.濃度為4、8、16、25μg/mL蜂膠均能明顯抑制細胞增殖相關的核抗原Ki67的表達,抑制強度和蜂膠濃度呈正相關性,同時一定濃度的蜂膠能夠誘導Egrl和P21的表達且表現出劑量相關性,隨著蜂膠的濃度增加而升高。 結論 (1)蜂膠提取物能夠刺激宮頸癌細胞炎癥因子IL-6、IL-8、IL-10和TNF-a及NO的分泌； (2)蜂膠提取物活菌顯著下調iNOS-Luc、PTGS-2-Luc和IL-8-Luc的表達,初步顯示與NF-κB家族蛋白有關； (3)經蜂膠誘導Hela細胞后,早期生長反應因子Egrl和細胞周期抑制基因p21在mRNA和蛋白水平上均有所升高,以及細胞增殖基因Ki67在mRNA水平上有所降低。表明這些細胞因子可能也參與了誘導Hela細胞分化的過程當中。
[Abstract]:Background and objective Cervical cancer is one of the malignant tumors with high mortality in female reproductive system, and its incidence tends to rise slowly. The purpose of this study was to explore the effect of propolis on cervical cancer cell line Hela, by selecting different concentrations of propolis extract, and to provide theoretical guidance for clinical treatment. Methods 1. The effects of propolis on cytokine secretion in Hela were detected by IHC assay. 2. The effects of propolis on inflammatory factors in Hela cells were systematically studied from DNA transcriptional regulation, RNA and protein levels. 3. The effects of propolis on the expression of Egr1,p21 and Ki67 related genes related to differentiation and proliferation of Hela cells were studied at RNA and protein levels. Propolis at concentrations of 4, 8 and 16, 25 渭 g / mL significantly affected the secretion of cytokines such as IL-8,TNF- 偽 and IL-10 in Hela cells, and it was found that when the concentration was above 16 渭 g / mL, the secretion of NO by macrophages was more than 7 times higher than that in the control group. 2. Propolis at concentrations of 4, 8 and 16, 25 渭 g / mL could regulate the expression of inflammatory factors iNOS,PTGS-2 and IL-8 in Hela cells at the level of DNA transcription, RNA and protein, which was shown to play a role through NF- kappa B cell signaling pathway. 3. When the concentration was 4, 8, 16, 25 渭 g / mL propolis could significantly inhibit the expression of nuclear antigen Ki67 related to cell proliferation, and the inhibitory intensity was positively correlated with the concentration of propolis. At the same time, a certain concentration of propolis could induce the expression of Egrl and P21 in a dose-dependent manner. It increases with the increase of propolis concentration. Conclusion (1) Propolis extract can stimulate the secretion of inflammatory factors IL-6,IL-8,IL-10, TNF-a and NO in cervical cancer cells. (2) the expression of iNOS-Luc,PTGS-2-Luc and IL-8-Luc was significantly down-regulated by living propolis extract, which was related to NF- kappa B family protein. (3) after Hela cells were induced by propolis, the early growth response factor Egrl and cell cycle inhibitor gene p21 increased at mRNA and protein levels, while the cell proliferation gene Ki67 decreased at mRNA level. These cytokines may also be involved in the differentiation of Hela cells.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.33

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