不同葡萄糖浓度在体外影响卵巢癌细胞生长与代谢的机制研究
发布时间：2018-12-08 10:17
【摘要】：背景作为危害人类健康的一大危险因素,肥胖不仅可以引起心血管的疾病,而且最近研究表明,肥胖与一些常见的恶性肿瘤的发生有关。但目前关于卵巢癌和肥胖的研究相对较少,而且目前尚无明确的统一定论。高血糖是营养过剩的一种独特的病理特征,而且是促进肿瘤发生发展的一个独立危险因子。葡萄糖作为维持细胞存活的一种重要的能量物质,有大量的研究表明葡萄糖的剥夺可能是一种有效的抗肿瘤的治疗方式。尽管在不同的恶性肿瘤中都有关于糖代谢的研究,但是对于卵巢癌细胞的生长作用机制和作用仍然有分歧。目的为了更好的了解糖代谢和卵巢癌细胞的关系,在我们的研究中,我们在体外配制不同的葡萄糖浓度的培养基,分别模拟无糖、低血糖浓度,正常血糖浓度及高血糖浓度的体内环境。在培养一定时间之后,拟观察不同的条件下对于卵巢癌SKOV3和A2780细胞的增殖、凋亡、细胞周期、活性氧以及相关信号通路的影响。方法在体外培养卵巢癌细胞株A2780、SKOV3,分别以葡萄糖浓度为0mmol/L、1mmol/L、5.5mmol/L、25.5mmol/L的培养条件,体外模拟无糖、低糖、正常糖、高糖的血糖状态。然后对两株卵巢癌细胞用不同葡萄糖浓度的培养基培养48H之后,用cck-8法、PI染色法分别检测检测两株细胞增殖、周期的变化;用Annexin V/PI法和DCFH-DA荧光探针法检测两株卵巢癌细胞凋亡以及活性氧(ROS)的变化;通过Western蛋白印迹实验检测上述卵巢癌细胞在不同糖浓度培养条件下,PI3K/AKT/mTOR,AMPK信号转导通路及相关通路蛋白(AKT、mTOR、AMPK)的磷酸化形式。结果通过我们的实验,我们发现,在体外,高糖有利于促进卵巢癌SKOV3和A2780细胞的增殖。与此同时,低糖可以使p-AMPK(Thr172)的表达量上调,同时p-AKT(Thr308)以及p-mTOR的表达量下调,低糖可以使卵巢癌细胞的G1期阻滞,而且可以使细胞的早期凋亡率增加。结论我们以此推测,通过控制体内的血糖稳定或者特异性针对细胞的糖代谢过程进行靶向治疗或许可以为卵巢癌的治疗提供了一种新的方向。
[Abstract]:Background as a major risk factor to human health obesity not only can cause cardiovascular diseases but also recent studies have shown that obesity is associated with the occurrence of some common malignant tumors. But there are relatively few studies on ovarian cancer and obesity, and there is no clear consensus. Hyperglycemia is a unique pathological feature of overnourishment and an independent risk factor for tumor development. Glucose is an important energy substance to maintain cell survival. A large number of studies have shown that glucose deprivation may be an effective antitumor therapy. Although glucose metabolism has been studied in different malignant tumors, there are still differences on the mechanism and role of ovarian cancer cell growth. Objective to better understand the relationship between glucose metabolism and ovarian cancer cells. In our study, we prepared different glucose concentration media in vitro to simulate hypoglycemia and no glucose concentration, respectively. The internal environment of normal and hyperglycemic concentrations. The effects of different conditions on the proliferation, apoptosis, cell cycle, reactive oxygen species (Ros) and related signaling pathways of ovarian cancer SKOV3 and A2780 cells were studied after a certain period of culture. Methods Ovarian cancer cell line A2780 SKOV3 was cultured in vitro under the condition of glucose concentration of 0 mmol / L 1 mmol / L 5. 5 mmol 路L ~ (5) mmol / L ~ (25. 5) mmol 路L ~ (-1) 路L ~ (2.5) mmol / L to simulate the glucose state of sugar free, low sugar, normal sugar and high sugar in vitro. Then two ovarian cancer cells were cultured in different glucose concentration culture medium for 48H, then the proliferation and cycle of the two ovarian cancer cells were detected by cck-8 method and PI staining method. Apoptosis and reactive oxygen species (Ros) (ROS) were detected by Annexin V/PI assay and DCFH-DA fluorescence probe assay. The phosphorylation of PI3K/AKT/mTOR,AMPK signal transduction pathway and related pathway protein (AKT,mTOR,AMPK) in ovarian cancer cells under different glucose concentrations was detected by Western blotting assay. Results through our experiments, we found that high glucose could promote the proliferation of ovarian cancer SKOV3 and A2780 cells in vitro. At the same time, low glucose could up-regulate the expression of p-AMPK (Thr172), decrease the expression of p-AKT (Thr308) and p-mTOR. Low glucose could block the G1 phase of ovarian cancer cells and increase the early apoptosis rate of ovarian cancer cells. Conclusion We speculate that targeted therapy for ovarian cancer may provide a new direction for the treatment of ovarian cancer by controlling the stability of blood glucose in vivo or targeting the process of glucose metabolism in cells.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31
本文编号：2368175
[Abstract]:Background as a major risk factor to human health obesity not only can cause cardiovascular diseases but also recent studies have shown that obesity is associated with the occurrence of some common malignant tumors. But there are relatively few studies on ovarian cancer and obesity, and there is no clear consensus. Hyperglycemia is a unique pathological feature of overnourishment and an independent risk factor for tumor development. Glucose is an important energy substance to maintain cell survival. A large number of studies have shown that glucose deprivation may be an effective antitumor therapy. Although glucose metabolism has been studied in different malignant tumors, there are still differences on the mechanism and role of ovarian cancer cell growth. Objective to better understand the relationship between glucose metabolism and ovarian cancer cells. In our study, we prepared different glucose concentration media in vitro to simulate hypoglycemia and no glucose concentration, respectively. The internal environment of normal and hyperglycemic concentrations. The effects of different conditions on the proliferation, apoptosis, cell cycle, reactive oxygen species (Ros) and related signaling pathways of ovarian cancer SKOV3 and A2780 cells were studied after a certain period of culture. Methods Ovarian cancer cell line A2780 SKOV3 was cultured in vitro under the condition of glucose concentration of 0 mmol / L 1 mmol / L 5. 5 mmol 路L ~ (5) mmol / L ~ (25. 5) mmol 路L ~ (-1) 路L ~ (2.5) mmol / L to simulate the glucose state of sugar free, low sugar, normal sugar and high sugar in vitro. Then two ovarian cancer cells were cultured in different glucose concentration culture medium for 48H, then the proliferation and cycle of the two ovarian cancer cells were detected by cck-8 method and PI staining method. Apoptosis and reactive oxygen species (Ros) (ROS) were detected by Annexin V/PI assay and DCFH-DA fluorescence probe assay. The phosphorylation of PI3K/AKT/mTOR,AMPK signal transduction pathway and related pathway protein (AKT,mTOR,AMPK) in ovarian cancer cells under different glucose concentrations was detected by Western blotting assay. Results through our experiments, we found that high glucose could promote the proliferation of ovarian cancer SKOV3 and A2780 cells in vitro. At the same time, low glucose could up-regulate the expression of p-AMPK (Thr172), decrease the expression of p-AKT (Thr308) and p-mTOR. Low glucose could block the G1 phase of ovarian cancer cells and increase the early apoptosis rate of ovarian cancer cells. Conclusion We speculate that targeted therapy for ovarian cancer may provide a new direction for the treatment of ovarian cancer by controlling the stability of blood glucose in vivo or targeting the process of glucose metabolism in cells.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31
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