發(fā)布時間：2018-04-19 23:07

  本文選題：膠質(zhì)瘤 + FBP1��； 參考：《南通大學》2014年碩士論文

【摘要】：目的1.從組織水平研究FBP1在膠質(zhì)瘤患者中的表達情況,同時分析FBP1與c-Myc、增殖指標Ki-67及臨床病理特征的相關性,探討FBP1與膠質(zhì)瘤發(fā)生和發(fā)展的關系。2.從細胞水平驗證FBP1在膠質(zhì)瘤細胞增殖中的影響,并探討FBP1參與膠質(zhì)瘤細胞生長調(diào)控的機制。方法1.在組織水平,選取70例不同病理分級的膠質(zhì)瘤石蠟包埋標本、9例新鮮膠質(zhì)瘤標本和3例新鮮正常大腦標本。免疫組化和Western blot檢測FBP1、c-Myc及Ki-67的表達情況,應用SPSS13.0統(tǒng)計軟件分析數(shù)據(jù)。2.在細胞水平,膠質(zhì)瘤細胞經(jīng)血清饑餓釋放同步化分析和流式細胞儀分析檢測細胞周期,Western blot和免疫熒光分析檢測FBP1、c-Myc在細胞內(nèi)的表達以及細胞定位。應用FBP1小干擾質(zhì)粒作用于膠質(zhì)瘤細胞,CCK8和流式細胞儀等分析其對膠質(zhì)瘤細胞周期的影響,免疫熒光技術和Western blot檢測干擾膠質(zhì)瘤細胞中FBP1、c-Myc的表達及功能的影響。結果1.Western blot和免疫組化結果顯示,FBP1在不同的病理分級中表達有顯著差異(P=0.013),隨膠質(zhì)瘤惡性程度增高表達增加。FBP1表達與膠質(zhì)瘤患者的性別、年齡、Karnofsky評分(KPS)、腫瘤發(fā)生部位、手術方式、腫瘤直徑、血管密度及壞死無明顯相關性。應用Spearman等級相關分析,我們發(fā)現(xiàn)FBP1與c-Myc(γ=0.740;P=0.005)和Ki-67(γ=0.830;P=0.009)表達呈正相關,c-Myc與Ki-67的表達呈正相關(γ=0.732;P=0.000)。Kaplan-Meier分析發(fā)現(xiàn)FBP1低表達(0-3)患者的五年生存率明顯高于高表達(4-7)的患者(P=0.004);Cox比例風險模型多因素生存分析證實FBP1的表達具有獨立的預后意義(P=0.037)。2.通過血清饑餓釋放分析發(fā)現(xiàn)隨著血清釋放刺激膠質(zhì)瘤細胞的增殖,FBP1表達增加,并可能轉(zhuǎn)錄調(diào)節(jié)c-Myc表達增加。FBP1干擾質(zhì)粒抑制FBP1的表達后,c-Myc的表達水平降低,同時,膠質(zhì)瘤細胞周期阻滯,膠質(zhì)瘤細胞生長受阻。結論1.FBP1的表達與膠質(zhì)瘤的病理分級密切相關,并且FBP1與c-Myc和Ki-67表達呈正相關,提示FBP1表達異常,可促進細胞異常增殖,從而促進膠質(zhì)瘤的發(fā)生及發(fā)展。2.FBP1與膠質(zhì)瘤患者的預后密切相關,FBP1的蛋白表達水平可作為膠質(zhì)瘤預后評估的指標之一。3.FBP1轉(zhuǎn)錄影響c-Myc分子的表達,促進細胞周期的進程,具有潛在的促癌性。4.小干擾質(zhì)粒靶向干擾FBP1,導致FBP1表達降低,從而誘導細胞周期阻滯,最終抑制腫瘤生長,為膠質(zhì)瘤的基因治療提供了新的靶點。
[Abstract]:Objective 1. To study the expression of FBP1 in glioma patients at the tissue level, and to analyze the correlation between FBP1 and c-Myc, proliferation index Ki-67 and clinicopathological features, and to explore the relationship between FBP1 and the occurrence and development of glioma. To verify the effect of FBP1 on glioma cell proliferation at cell level and to explore the mechanism of FBP1 involved in glioma cell growth regulation. Method 1. At the histological level, 70 cases of paraffin embedded gliomas with different pathological grades were collected from 9 cases of fresh gliomas and 3 cases of fresh normal brain specimens. Immunohistochemistry and Western blot were used to detect the expression of c-Myc and Ki-67 in FBP1. The data were analyzed by SPSS13.0 software. At the cell level, the expression and localization of FBP1c-Myc in glioma cells were detected by serum starvation release synchronization analysis and flow cytometry, respectively, by Western blot and immunofluorescence analysis. The effect of FBP1 small interference plasmid on glioma cell cycle was analyzed by flow cytometry and CCK8. Immunofluorescence and Western blot were used to detect the expression and function of FBP1 c-Myc in glioma cells. Results the results of 1.Western blot and immunohistochemistry showed that there were significant differences in the expression of FBP1 in different pathological grades. The expression of FBP1 increased with the malignant degree of glioma. The expression of FBP1 was correlated with the sex, age and age of glioma patients. There was no significant correlation between tumor diameter, vascular density and necrosis. Applying Spearman rank correlation analysis, We found that there was a positive correlation between the expression of FBP1 and c-Myc (緯 -0.700.40) and Ki-67 (緯 -0.830) and the positive correlation between c-Myc and Ki-67. Kaplan-Meier analysis showed that the 5-year survival rate of patients with low expression of FBP1 was significantly higher than that of patients with high expression of 4-7 (P < 0.001). Multivariate survival analysis confirmed that the survival rate of patients with low expression of FBP1 was significantly higher than that of patients with high expression of 4-7. The multivariate survival model showed that c-Myc was positively correlated with the expression of c-Myc (緯 -0.732P 0.0000.000.Kaplan-Meier analysis). The expression of FBP1 has independent prognostic significance. The results of serum starvation release analysis showed that the expression of FBP1 in glioma cells increased with the release of serum, and the expression of c-Myc decreased after the inhibition of FBP1 expression by FBP1 interference plasmids. Glioma cell cycle arrest, glioma cell growth block. Conclusion the expression of 1.FBP1 is closely related to the pathological grade of gliomas, and the expression of FBP1 is positively correlated with the expression of c-Myc and Ki-67, suggesting that the abnormal expression of FBP1 can promote the abnormal proliferation of glioma cells. Therefore, the expression of FBP1 protein is closely related to the prognosis of glioma patients. 3. FBP1 transcription affects the expression of c-Myc molecules and promotes the progress of cell cycle. Have potential carcinogenicity. 4. The small interference plasmid interferes with FBP1, which leads to the decrease of FBP1 expression, thus induces cell cycle arrest, and finally inhibits tumor growth, which provides a new target for gene therapy of glioma.
【學位授予單位】：南通大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R739.41
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本文編號：1775133