本文選題：Notch1基因 + RNA��； 參考：《泰山醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的 探究Notch1在腎癌組織中的表達(dá)及與腎癌臨床病理之間的關(guān)聯(lián)。進(jìn)一步探究Notch1在腎透明細(xì)胞癌中的表達(dá)含量，利用小片段RNA(siRNA)干預(yù)方法沉默腎透明細(xì)胞癌中Notch1的表達(dá)，并觀測(cè)其對(duì)在腎癌Caki-1細(xì)胞增殖的意義。探索Notch1基因在腎癌發(fā)生發(fā)展中的臨床意義。 方法 搜集52例腎癌組織與30例非癌腎組織標(biāo)本，利用熒光實(shí)時(shí)定量PCR（RT-PCR）和免疫組化方式剖析正常腎組織與腎癌組織中Notch1基因的表達(dá)差別，分析Notch1與臨床病理參數(shù)的相關(guān)性聯(lián)系。人體正常HK-2腎小管上皮細(xì)胞和腎細(xì)胞癌786O細(xì)胞和Caki-1體外培養(yǎng)使用。經(jīng)過(guò)實(shí)時(shí)熒光定量PCR(RT-PCR)檢測(cè)Notch1mRNA和蛋白印跡法（Western blot）檢測(cè)蛋白的表達(dá)。設(shè)立特異性Notch1-siRNA干涉腎癌Caki-1細(xì)胞，用Lip2000轉(zhuǎn)染試劑轉(zhuǎn)染48時(shí)小，經(jīng)過(guò)熒光定量PCR和印跡蛋白法分別檢測(cè)Notch1mRNA和蛋白的表達(dá)含量不同，CCk-8與克隆試驗(yàn)了解其增殖情行的變化。 結(jié)果 Notch1于腎癌組織中的表現(xiàn)高于非腎癌腎組織（為80.8%比40.0%，P0.05)。Notch1的表達(dá)水平同樣與腎癌的遠(yuǎn)處轉(zhuǎn)移有關(guān)，發(fā)生轉(zhuǎn)移的高于未轉(zhuǎn)移的(P0.05)；直徑≥7cm的癌中的Notch1含量要高于直徑＜7cm的腫瘤(P0.05)。類似的，，Notch1于腎癌細(xì)胞株中的含量要高于正常腎小管細(xì)胞，具備統(tǒng)計(jì)學(xué)差異（P0.05)。干擾siRNA-notch1后可顯著降低Caki-1細(xì)胞中Notch1mRNA和蛋白的表達(dá)，具有統(tǒng)計(jì)學(xué)差異（P0.05)，也檢測(cè)出Caki-1細(xì)胞增殖本領(lǐng)的降低，差異也具備統(tǒng)計(jì)學(xué)差別（P0.05)。 結(jié)論 Notch1在腎癌中高表達(dá)大概與腎癌發(fā)生發(fā)展有相關(guān)性。同時(shí)，Notch1與腎癌的臨床病理分期分級(jí)和腫瘤大小均有關(guān)性。腎透亮細(xì)胞癌組織中Notch1基因高表達(dá)，Notch1信號(hào)通路的下調(diào)可抑制腎細(xì)胞癌Caki-1細(xì)胞發(fā)生，調(diào)控腎細(xì)胞癌，對(duì)腎細(xì)胞癌的診斷和治療有一定的指導(dǎo)意義。
[Abstract]:Objective to investigate the expression of Notch1 in renal cell carcinoma (RCC) and its relationship with clinicopathology. To further investigate the expression of Notch1 in renal clear cell carcinoma (RCC), the expression of Notch1 in renal clear cell carcinoma (RCC) was silenced by small fragment RNA-siRNAs intervention, and the significance of Notch1 expression in Caki-1 cell proliferation was observed. To explore the clinical significance of Notch1 gene in the development of renal cell carcinoma. Methods 52 cases of renal carcinoma and 30 cases of non-cancerous renal tissues were collected. The expression of Notch1 gene in normal renal tissues and renal cell carcinoma tissues was analyzed by real-time quantitative PCR and immunohistochemistry. To analyze the correlation between Notch1 and clinicopathological parameters. Normal HK-2 renal tubular epithelial cells and renal cell carcinoma 786O cells and Caki-1 were cultured in vitro. The expression of Notch1 mRNA and Western blot were detected by real-time fluorescence quantitative PCR RT-PCR. The specific Notch1-siRNA interference cell line Caki-1 was established and transfected with Lip2000 transfection reagent for 48 hours. The expression of Notch1 mRNA and protein was detected by fluorescence quantitative PCR and blotting method respectively. Results the expression of Notch1 in renal cell carcinoma was higher than that in non-renal cell carcinoma (80.8% vs 40.0%). The expression level of Notch1 was also related to the distant metastasis of renal carcinoma. The content of Notch1 in carcinoma with diameter 鈮

本文編號(hào)：2043971