FBP1与胶质瘤的相关性研究

发布时间：2018-04-19 23:07

  本文选题：胶质瘤 + FBP1　； 参考：《南通大学》2014年硕士论文

【摘要】：目的1.从组织水平研究FBP1在胶质瘤患者中的表达情况,同时分析FBP1与c-Myc、增殖指标Ki-67及临床病理特征的相关性,探讨FBP1与胶质瘤发生和发展的关系。2.从细胞水平验证FBP1在胶质瘤细胞增殖中的影响,并探讨FBP1参与胶质瘤细胞生长调控的机制。方法1.在组织水平,选取70例不同病理分级的胶质瘤石蜡包埋标本、9例新鲜胶质瘤标本和3例新鲜正常大脑标本。免疫组化和Western blot检测FBP1、c-Myc及Ki-67的表达情况,应用SPSS13.0统计软件分析数据。2.在细胞水平,胶质瘤细胞经血清饥饿释放同步化分析和流式细胞仪分析检测细胞周期,Western blot和免疫荧光分析检测FBP1、c-Myc在细胞内的表达以及细胞定位。应用FBP1小干扰质粒作用于胶质瘤细胞,CCK8和流式细胞仪等分析其对胶质瘤细胞周期的影响,免疫荧光技术和Western blot检测干扰胶质瘤细胞中FBP1、c-Myc的表达及功能的影响。结果1.Western blot和免疫组化结果显示,FBP1在不同的病理分级中表达有显著差异(P=0.013),随胶质瘤恶性程度增高表达增加。FBP1表达与胶质瘤患者的性别、年龄、Karnofsky评分(KPS)、肿瘤发生部位、手术方式、肿瘤直径、血管密度及坏死无明显相关性。应用Spearman等级相关分析,我们发现FBP1与c-Myc(γ=0.740;P=0.005)和Ki-67(γ=0.830;P=0.009)表达呈正相关,c-Myc与Ki-67的表达呈正相关(γ=0.732;P=0.000)。Kaplan-Meier分析发现FBP1低表达(0-3)患者的五年生存率明显高于高表达(4-7)的患者(P=0.004);Cox比例风险模型多因素生存分析证实FBP1的表达具有独立的预后意义(P=0.037)。2.通过血清饥饿释放分析发现随着血清释放刺激胶质瘤细胞的增殖,FBP1表达增加,并可能转录调节c-Myc表达增加。FBP1干扰质粒抑制FBP1的表达后,c-Myc的表达水平降低,同时,胶质瘤细胞周期阻滞,胶质瘤细胞生长受阻。结论1.FBP1的表达与胶质瘤的病理分级密切相关,并且FBP1与c-Myc和Ki-67表达呈正相关,提示FBP1表达异常,可促进细胞异常增殖,从而促进胶质瘤的发生及发展。2.FBP1与胶质瘤患者的预后密切相关,FBP1的蛋白表达水平可作为胶质瘤预后评估的指标之一。3.FBP1转录影响c-Myc分子的表达,促进细胞周期的进程,具有潜在的促癌性。4.小干扰质粒靶向干扰FBP1,导致FBP1表达降低,从而诱导细胞周期阻滞,最终抑制肿瘤生长,为胶质瘤的基因治疗提供了新的靶点。
[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 (纬 -0.700.40) and Ki-67 (纬 -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 (纬 -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.
【学位授予单位】：南通大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41
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本文编号：1775133