本文選題：宮頸癌　切入點(diǎn)：葉酸　出處：《山西醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的:宮頸癌作為全球女性最為常見的婦科惡性腫瘤之一,發(fā)病率和死亡率呈逐年上升趨勢(shì)。葉酸是機(jī)體必需的微量營(yíng)養(yǎng)素,相關(guān)流行病學(xué)證據(jù)亦表明葉酸與宮頸癌的發(fā)生有著密切的關(guān)系。自噬是細(xì)胞死亡的一種獨(dú)特方式,細(xì)胞自噬在對(duì)抗腫瘤生成方面也扮演了作用的角色。但對(duì)于宮頸癌、自噬、葉酸這三者間關(guān)系的研究卻尚未涉及,因此它們的關(guān)系未明確。本實(shí)驗(yàn)旨在探討葉酸對(duì)宮頸癌SiHa細(xì)胞生物學(xué)功能及對(duì)自噬基因βeclin-1表達(dá)的影響。方法:1.常規(guī)條件下培養(yǎng)SiHa細(xì)胞,利用1ug/mL(對(duì)照組)、10ug/ml、100ug/ml、500ug/ml、1000ug/ml的葉酸進(jìn)行干預(yù),觀察并記錄細(xì)胞的形態(tài)變化和生長(zhǎng)速度。2.利用1ug/mL(對(duì)照組)、10ug/ml、100ug/ml、500ug/ml、1000ug/ml的葉酸進(jìn)行干預(yù)后以CCK8檢測(cè)宮頸癌SiHa細(xì)胞的增殖活性,劃痕實(shí)驗(yàn)觀察細(xì)胞的遷移能力及流式細(xì)胞儀分析細(xì)胞的凋亡情況,采用Western bloting檢測(cè)自噬蛋白βeclin-1表達(dá)水平。結(jié)果:1.與對(duì)照組比較,葉酸濃度達(dá)到1000ug/ml時(shí)細(xì)胞形態(tài)發(fā)生明顯變化,當(dāng)實(shí)驗(yàn)組葉酸濃度在100ug/ml及以上時(shí),與對(duì)照組相比,實(shí)驗(yàn)組細(xì)胞總數(shù)明顯降低。2.CCK8細(xì)胞增殖實(shí)驗(yàn)顯示在不同葉酸濃度干預(yù)下,除了10ug/ml葉酸的實(shí)驗(yàn)組與對(duì)照組相比P=0.650無(wú)差異無(wú)統(tǒng)計(jì)學(xué)意義外,100ug/ml、500ug/ml、1000ug/ml濃度的葉酸細(xì)胞增殖與對(duì)照組相比均有統(tǒng)計(jì)學(xué)意義,P值分別為0.041、0.001、0.001。3.劃痕實(shí)驗(yàn)表明10ug/ml的葉酸實(shí)驗(yàn)組劃痕愈合面積與對(duì)照組比較差異不明顯。100ug/ml、500ug/ml、1000ug/ml濃度的葉酸實(shí)驗(yàn)組與對(duì)照組相比劃痕愈合面積減少,且隨著葉酸濃度的增加愈合面積減少愈發(fā)明顯。4.流式細(xì)胞儀分析葉酸對(duì)宮頸癌細(xì)胞凋亡的影響,結(jié)果顯示10ug/ml的葉酸實(shí)驗(yàn)組細(xì)胞的凋亡率與對(duì)照組比較P=0.580,差異無(wú)統(tǒng)計(jì)學(xué)意義外,100ug/ml、500ug/ml、1000ug/ml濃度的葉酸細(xì)胞的凋亡率與對(duì)照組相比均有統(tǒng)計(jì)學(xué)意義,P值分別為0.020、0.001、0.001。5.Wesrern Bloting檢測(cè)葉酸對(duì)宮頸癌細(xì)胞自噬基因βeclin-1蛋白表達(dá)的作用。實(shí)驗(yàn)組10ug/ml、100ug/ml、500ug/ml、1000ug/ml濃度的葉酸實(shí)驗(yàn)組βeclin-1蛋白表達(dá)水平與對(duì)照組相比差異均有統(tǒng)計(jì)學(xué)意義P值分別為0.014、0.003、0.001、0.001。結(jié)論:1.高濃度葉酸抑制宮頸癌SiHa細(xì)胞的增殖、遷移,促進(jìn)宮頸癌SiHa細(xì)胞的凋亡。2.高濃度葉酸促進(jìn)宮頸癌自噬蛋白βeclin-1的表達(dá)。
[Abstract]:Objective: cervical cancer is one of the most common gynecological malignancies in the world.Folic acid is an essential micronutrient in the body. Relevant epidemiological evidence also shows that folic acid is closely related to the occurrence of cervical cancer.Autophagy is a unique way of cell death. Autophagy also plays a role in fighting tumor formation.However, the relationship among cervical cancer, autophagy and folic acid has not been studied, so their relationship is not clear.The aim of this study was to investigate the effects of folic acid on the biological function of cervical cancer SiHa cells and the expression of autophagy gene 尾 eclin-1.Method 1: 1.SiHa cells were cultured under conventional conditions and treated with folic acid of 10ugrmL (control group). The morphological changes and growth rate of the cells were observed and recorded.The result is 1: 1.When the concentration of folic acid reached 1000ug/ml, the morphology of the cells changed obviously. When the concentration of folic acid in the experimental group was 100ug/ml and above, it was compared with the control group.The total number of cells in the experimental group was significantly decreased. 2. CCK8 cell proliferation assay showed that the cells were treated with different folic acid concentrations.There was no significant difference in the proliferation of folic acid cells between the experimental group and the control group except that of the control group. The proliferation of folic acid cells at the concentration of 100U / ml / ml / ml / ml was significantly higher than that of the control group (P = 0.041 / 0.001 / ml, P = 0.001.3respectively).Scratch test showed that there was no significant difference in scratch healing area between the folic acid experimental group of 10ug/ml and the control group. Compared with the control group, the area of scratch healing in the folic acid experimental group with the concentration of 100ugml / ml was less than that in the control group, and with the increase of folic acid concentration, the healing area of folic acid group was decreased more obviously than that of the control group.The effect of folic acid on apoptosis of cervical cancer cells was analyzed by flow cytometry.The results showed that the apoptotic rate of the folic acid experimental group of 10ug/ml was 0.580 compared with that of the control group. There was no significant difference between the two groups. The apoptosis rate of the folic acid cells at the concentration of 100U / ml / ml was 0.020 鹵0.001 / ml and 0.001.5.Wesrern Bloting was used to detect the apoptosis rate of the folic acid cells in the control group and the control group respectively (P = 0.020, P = 0.020, P = 0.001.5.Wesrern Bloting).Effect of 尾 eclin-1 protein on autophagy gene in cervical cancer cells.Compared with the control group, the expression level of 尾 eclin-1 protein in the experimental group (10ugr / ml) was significantly higher than that in the control group (P = 0.014, 0.003, P = 0.001, P = 0.001, respectively), and the expression level of 尾 eclin-1 protein in the experimental group was significantly higher than that in the control group (P < 0.05).Conclusion 1.High concentration folic acid inhibited the proliferation and migration of cervical cancer SiHa cells and promoted the apoptosis of cervical cancer SiHa cells.High concentration folic acid promoted the expression of autophagy protein 尾 eclin-1 in cervical cancer.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R737.33

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1 王燕琴;葉酸對(duì)宮頸癌SiHa細(xì)胞生物學(xué)特性及βeclin-1蛋白表達(dá)影響的初步研究[D];山西醫(yī)科大學(xué);2016年

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本文編號(hào)：1695511