RHEB在肾透明细胞癌中表达及其临床意义的研究

发布时间：2018-06-27 18:19

本文选题：RHEB + 肾透明细胞癌　；参考：《宁夏医科大学》2017年硕士论文

【摘要】：目的:1、探讨RHEB在肾透明细胞癌(RCCC)的癌组织及癌旁非肿瘤组织中表达情况,并分析与其临床意义。2、使用特异性的RHEB-shRNA干扰序列转染人肾癌细胞株ACHN,诱导RHEB基因沉默,进一步观察其对ACHN细胞基因表达的影响。方法:1、根据课题组前期已完成通过收集肾透明细胞癌的新鲜组织样本提取RNA应用高通量测序技术分析转移组样本和未转移组样本的差异基因表达结果,并查阅国内外相关文献,最终确定RHEB为本次研究的靶基因。2、通过免疫组织化学法研究82例人肾透明细胞癌及癌旁非肿瘤组织中的RHEB蛋白表达情况及其临床意义。3、通过实时荧光定量PCR和免疫印迹试验两种技术方法来分析转染shRHEB的ACHN细胞中的RHEB及其蛋白表达水平。结果:1.免疫组化方法检测RHEB在RCCC组织中的阳性表达率为91.46%(75/82),在癌旁非肿瘤组织的阳性表达率为30.49%(25/82)。经卡方检验,差异具有统计学意义(χ2=64.06,P=0.000)。RHEB在局限性肾癌、局部进展性肾癌及转移性肾癌之间表达的阳性等级具有显著差异(P=0.044),且随着肿瘤临床分期的升高,RHEB表达的阳性等级呈逐渐上升的趋势;肾透明细胞不同病理分级的癌组织中,RHEB表达的阳性等级无显著差异(P=0.166);RHEB阳性表达与性别(P0.05)、年龄(P0.05)、肿瘤直径大小(P0.05)均没有相关性。2.通过实时定量实验检测RHEB基因在转染shRHEB的ACHN细胞中的表达量与对照组ACHN细胞相比,其表达量显著下降(0.382±0.015,P=0.000);Western blot印迹实验检测在转染shRHEB的ACHN细胞中的RHEB蛋白的表达,和对照ACHN细胞相比,其表达量也显著下降(P=0.000)。结论:RHEB可能与肾透明细胞癌的发生、发展有着一定的关系。
[Abstract]:Objective to investigate the expression of RHEB in human renal clear cell carcinoma (RCCC) and its adjacent non-tumor tissues, and to analyze the clinical significance of RHEB gene silencing in human renal cell carcinoma cell line ACHN by using specific RHEB-shRNA interference sequence. To investigate the effect of ACHN on gene expression in ACHN cells. Methods: 1. According to the results of RNA extraction by collecting fresh tissue samples from clear cell carcinoma of kidney, using high-throughput sequencing technique, we analyzed the differentially expressed genes between the samples of transfer group and non-metastatic group, and consulted the relevant literatures at home and abroad. RHEB was identified as the target gene of this study. The expression of RHEB protein and its clinical significance in 82 cases of human renal clear cell carcinoma and its adjacent non-tumor tissues were studied by immunohistochemical method. The expression of RHEB protein and its clinical significance were studied by real-time fluorescence quantitative PCR and immunoassay. The expression of RHEB and its protein in shRHEB transfected ACHN cells was analyzed by two methods of epidemic blotting. The result is 1: 1. The positive expression rate of RHEB was 91.46% (75 / 82) in RCCC and 30.49% (25 / 82) in non-tumor tissues. By chi-square test, the difference was statistically significant (蠂 2 / 64.06P = 0.000). There was significant difference in the positive grade of RHEB expression between local progressive renal cell carcinoma and metastatic renal cell carcinoma (P0. 044), and the positive grade of RHEB expression gradually increased with the increase of clinical stage of tumor. There was no significant difference between the positive grade of RHEB expression and sex (P0.05), age (P0.05) and tumor diameter (P0.05) in renal clear cell carcinoma tissues with different pathological grades (P0. 166). There was no correlation between RHEB expression and sex (P0.05), tumor diameter (P0.05). The expression of RHEB gene in shRHEB transfected ACHN cells was detected by real-time quantitative assay. The expression of RHEB gene in shRHEB transfected ACHN cells was significantly decreased (0.382 卤0.015 P0. 000). Western blot was used to detect the expression of RHEB protein in shRHEB transfected ACHN cells. Compared with the control ACHN cells, the expression of ACHN was significantly decreased (P0. 000). Conclusion: RHEB may be associated with the development of renal clear cell carcinoma.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.11

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