蜂胶对宫颈癌Hela细胞作用的研究
发布时间：2019-05-10 08:58
【摘要】：背景和目的 宫颈癌是女性生殖系统中病死率较高的恶性肿瘤之一,其发病率呈缓慢上升的趋势。本研究欲通过选用不同浓度蜂胶提取物处理宫颈癌细胞Hela,探讨蜂胶对其影响,为临床治疗提供理论指导。 方法 1、采用IHC法检测蜂胶对Hela中细胞因子分泌的影响； 2、从DNA转录调控,RNA和蛋白水平系统研究蜂胶对Hela细胞中炎症因子的影响； 3、从RNA和蛋白水平研究蜂胶对Hela细胞分化增殖相关基因Egr1、p21和Ki67表达的影响； 结果 1.浓度4、8、16、25μg/mL的蜂胶可以显著影响Hela细胞中IL-8、TNF-α和IL-10等细胞因子的分泌,同时发现浓度达到16μg/mL以上时,对巨噬细胞刺激分泌NO可达对照组的7倍以上。 2.浓度4、8、16、25μg/mL的蜂胶能从DNA转录调控,RNA和蛋白水平上调节Hela细胞中炎症因子iNOS、PTGS-2和IL-8的表达,初步显示通过NF-κB细胞信号通路起作用。 3.浓度为4、8、16、25μg/mL蜂胶均能明显抑制细胞增殖相关的核抗原Ki67的表达,抑制强度和蜂胶浓度呈正相关性,同时一定浓度的蜂胶能够诱导Egrl和P21的表达且表现出剂量相关性,随着蜂胶的浓度增加而升高。 结论 (1)蜂胶提取物能够刺激宫颈癌细胞炎症因子IL-6、IL-8、IL-10和TNF-a及NO的分泌； (2)蜂胶提取物活菌显著下调iNOS-Luc、PTGS-2-Luc和IL-8-Luc的表达,初步显示与NF-κB家族蛋白有关； (3)经蜂胶诱导Hela细胞后,早期生长反应因子Egrl和细胞周期抑制基因p21在mRNA和蛋白水平上均有所升高,以及细胞增殖基因Ki67在mRNA水平上有所降低。表明这些细胞因子可能也参与了诱导Hela细胞分化的过程当中。
[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- 伪 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.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33
[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- 伪 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.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33
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