【摘要】：背景作為危害人類健康的一大危險(xiǎn)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(jiàn)的惡性腫瘤的發(fā)生有關(guān)。但目前關(guān)于卵巢癌和肥胖的研究相對(duì)較少,而且目前尚無(wú)明確的統(tǒng)一定論。高血糖是營(yíng)養(yǎng)過(guò)剩的一種獨(dú)特的病理特征,而且是促進(jìn)腫瘤發(fā)生發(fā)展的一個(gè)獨(dú)立危險(xiǎn)因子。葡萄糖作為維持細(xì)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(guān)于糖代謝的研究,但是對(duì)于卵巢癌細(xì)胞的生長(zhǎng)作用機(jī)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(xì)胞的關(guān)系,在我們的研究中,我們?cè)隗w外配制不同的葡萄糖濃度的培養(yǎng)基,分別模擬無(wú)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(shí)間之后,擬觀察不同的條件下對(duì)于卵巢癌SKOV3和A2780細(xì)胞的增殖、凋亡、細(xì)胞周期、活性氧以及相關(guān)信號(hào)通路的影響。方法在體外培養(yǎng)卵巢癌細(xì)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(wú)糖、低糖、正常糖、高糖的血糖狀態(tài)。然后對(duì)兩株卵巢癌細(xì)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(cè)檢測(cè)兩株細(xì)胞增殖、周期的變化;用Annexin V/PI法和DCFH-DA熒光探針?lè)z測(cè)兩株卵巢癌細(xì)胞凋亡以及活性氧(ROS)的變化;通過(guò)Western蛋白印跡實(shí)驗(yàn)檢測(cè)上述卵巢癌細(xì)胞在不同糖濃度培養(yǎng)條件下,PI3K/AKT/mTOR,AMPK信號(hào)轉(zhuǎn)導(dǎo)通路及相關(guān)通路蛋白(AKT、mTOR、AMPK)的磷酸化形式。結(jié)果通過(guò)我們的實(shí)驗(yàn),我們發(fā)現(xiàn),在體外,高糖有利于促進(jìn)卵巢癌SKOV3和A2780細(xì)胞的增殖。與此同時(shí),低糖可以使p-AMPK(Thr172)的表達(dá)量上調(diào),同時(shí)p-AKT(Thr308)以及p-mTOR的表達(dá)量下調(diào),低糖可以使卵巢癌細(xì)胞的G1期阻滯,而且可以使細(xì)胞的早期凋亡率增加。結(jié)論我們以此推測(cè),通過(guò)控制體內(nèi)的血糖穩(wěn)定或者特異性針對(duì)細(xì)胞的糖代謝過(guò)程進(jìn)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(xué)位授予單位】：濟(jì)南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.31

【參考文獻(xiàn)】

相關(guān)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(guān)系在腫瘤方面的研究進(jìn)展[J];現(xiàn)代腫瘤醫(yī)學(xué);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(shí)用醫(yī)學(xué)雜志;2012年22期

4 歐陽(yáng)高亮,李祺福,洪水根;細(xì)胞凋亡與腫瘤的發(fā)生發(fā)展和治療[J];國(guó)外醫(yī)學(xué)(腫瘤學(xué)分冊(cè));2000年05期

相關(guān)碩士學(xué)位論文 前1條

1 孫永杰;體重指數(shù)與惡性腫瘤之間相關(guān)性的分析[D];新鄉(xiāng)醫(yī)學(xué)院;2015年

，

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