本文選題：RHEB + 腎透明細(xì)胞癌��； 參考：《寧夏醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:1、探討RHEB在腎透明細(xì)胞癌(RCCC)的癌組織及癌旁非腫瘤組織中表達(dá)情況,并分析與其臨床意義。2、使用特異性的RHEB-shRNA干擾序列轉(zhuǎn)染人腎癌細(xì)胞株ACHN,誘導(dǎo)RHEB基因沉默,進(jìn)一步觀察其對(duì)ACHN細(xì)胞基因表達(dá)的影響。方法:1、根據(jù)課題組前期已完成通過(guò)收集腎透明細(xì)胞癌的新鮮組織樣本提取RNA應(yīng)用高通量測(cè)序技術(shù)分析轉(zhuǎn)移組樣本和未轉(zhuǎn)移組樣本的差異基因表達(dá)結(jié)果,并查閱國(guó)內(nèi)外相關(guān)文獻(xiàn),最終確定RHEB為本次研究的靶基因。2、通過(guò)免疫組織化學(xué)法研究82例人腎透明細(xì)胞癌及癌旁非腫瘤組織中的RHEB蛋白表達(dá)情況及其臨床意義。3、通過(guò)實(shí)時(shí)熒光定量PCR和免疫印跡試驗(yàn)兩種技術(shù)方法來(lái)分析轉(zhuǎn)染shRHEB的ACHN細(xì)胞中的RHEB及其蛋白表達(dá)水平。結(jié)果:1.免疫組化方法檢測(cè)RHEB在RCCC組織中的陽(yáng)性表達(dá)率為91.46%(75/82),在癌旁非腫瘤組織的陽(yáng)性表達(dá)率為30.49%(25/82)。經(jīng)卡方檢驗(yàn),差異具有統(tǒng)計(jì)學(xué)意義(χ2=64.06,P=0.000)。RHEB在局限性腎癌、局部進(jìn)展性腎癌及轉(zhuǎn)移性腎癌之間表達(dá)的陽(yáng)性等級(jí)具有顯著差異(P=0.044),且隨著腫瘤臨床分期的升高,RHEB表達(dá)的陽(yáng)性等級(jí)呈逐漸上升的趨勢(shì);腎透明細(xì)胞不同病理分級(jí)的癌組織中,RHEB表達(dá)的陽(yáng)性等級(jí)無(wú)顯著差異(P=0.166);RHEB陽(yáng)性表達(dá)與性別(P0.05)、年齡(P0.05)、腫瘤直徑大小(P0.05)均沒(méi)有相關(guān)性。2.通過(guò)實(shí)時(shí)定量實(shí)驗(yàn)檢測(cè)RHEB基因在轉(zhuǎn)染shRHEB的ACHN細(xì)胞中的表達(dá)量與對(duì)照組ACHN細(xì)胞相比,其表達(dá)量顯著下降(0.382±0.015,P=0.000);Western blot印跡實(shí)驗(yàn)檢測(cè)在轉(zhuǎn)染shRHEB的ACHN細(xì)胞中的RHEB蛋白的表達(dá),和對(duì)照ACHN細(xì)胞相比,其表達(dá)量也顯著下降(P=0.000)。結(jié)論:RHEB可能與腎透明細(xì)胞癌的發(fā)生、發(fā)展有著一定的關(guān)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 鹵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.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R737.11

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