不同葡萄糖濃度在體外影響卵巢癌細胞生長與代謝的機制研究
發(fā)布時間：2018-12-08 10:17
【摘要】：背景作為危害人類健康的一大危險因素,肥胖不僅可以引起心血管的疾病,而且最近研究表明,肥胖與一些常見的惡性腫瘤的發(fā)生有關。但目前關于卵巢癌和肥胖的研究相對較少,而且目前尚無明確的統(tǒng)一定論。高血糖是營養(yǎng)過剩的一種獨特的病理特征,而且是促進腫瘤發(fā)生發(fā)展的一個獨立危險因子。葡萄糖作為維持細胞存活的一種重要的能量物質(zhì),有大量的研究表明葡萄糖的剝奪可能是一種有效的抗腫瘤的治療方式。盡管在不同的惡性腫瘤中都有關于糖代謝的研究,但是對于卵巢癌細胞的生長作用機制和作用仍然有分歧。目的為了更好的了解糖代謝和卵巢癌細胞的關系,在我們的研究中,我們在體外配制不同的葡萄糖濃度的培養(yǎng)基,分別模擬無糖、低血糖濃度,正常血糖濃度及高血糖濃度的體內(nèi)環(huán)境。在培養(yǎng)一定時間之后,擬觀察不同的條件下對于卵巢癌SKOV3和A2780細胞的增殖、凋亡、細胞周期、活性氧以及相關信號通路的影響。方法在體外培養(yǎng)卵巢癌細胞株A2780、SKOV3,分別以葡萄糖濃度為0mmol/L、1mmol/L、5.5mmol/L、25.5mmol/L的培養(yǎng)條件,體外模擬無糖、低糖、正常糖、高糖的血糖狀態(tài)。然后對兩株卵巢癌細胞用不同葡萄糖濃度的培養(yǎng)基培養(yǎng)48H之后,用cck-8法、PI染色法分別檢測檢測兩株細胞增殖、周期的變化;用Annexin V/PI法和DCFH-DA熒光探針法檢測兩株卵巢癌細胞凋亡以及活性氧(ROS)的變化;通過Western蛋白印跡實驗檢測上述卵巢癌細胞在不同糖濃度培養(yǎng)條件下,PI3K/AKT/mTOR,AMPK信號轉導通路及相關通路蛋白(AKT、mTOR、AMPK)的磷酸化形式。結果通過我們的實驗,我們發(fā)現(xiàn),在體外,高糖有利于促進卵巢癌SKOV3和A2780細胞的增殖。與此同時,低糖可以使p-AMPK(Thr172)的表達量上調(diào),同時p-AKT(Thr308)以及p-mTOR的表達量下調(diào),低糖可以使卵巢癌細胞的G1期阻滯,而且可以使細胞的早期凋亡率增加。結論我們以此推測,通過控制體內(nèi)的血糖穩(wěn)定或者特異性針對細胞的糖代謝過程進行靶向治療或許可以為卵巢癌的治療提供了一種新的方向。
[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.
【學位授予單位】：濟南大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R737.31
本文編號：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.
【學位授予單位】：濟南大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R737.31
【參考文獻】
相關期刊論文 前4條
1 鄧正亭;李ng健;程彬彬;;自噬與凋亡的相互關系在腫瘤方面的研究進展[J];現(xiàn)代腫瘤醫(yī)學;2015年10期
2 JI Cheng Ye;CHEN Tian Jiao;;Empirical Changes in the Prevalence of Overweight and Obesity among Chinese Students from 1985 to 2010 and Corresponding Preventive Strategies[J];Biomedical and Environmental Sciences;2013年01期
3 劉洋;許新華;;糖代謝異常與腫瘤發(fā)生發(fā)展[J];實用醫(yī)學雜志;2012年22期
4 歐陽高亮,李祺福,洪水根;細胞凋亡與腫瘤的發(fā)生發(fā)展和治療[J];國外醫(yī)學(腫瘤學分冊);2000年05期
相關碩士學位論文 前1條
1 孫永杰;體重指數(shù)與惡性腫瘤之間相關性的分析[D];新鄉(xiāng)醫(yī)學院;2015年
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