本文選題：SATB1　切入點(diǎn)：腎細(xì)胞癌　出處：《華中科技大學(xué)》2014年博士論文

【摘要】：目的：探討核轉(zhuǎn)錄因子SATB1在不同腎癌細(xì)胞系中的表達(dá)情況,以及SATB1基因表達(dá)對腎癌細(xì)胞增殖、侵襲能力的影響。 方法：分別應(yīng)用實(shí)時(shí)定量PCR、Western blotting與免疫熒光染色法檢測不同腎癌細(xì)胞系中SATB1的表達(dá)情況。在構(gòu)建靶向沉默SATB1的pGenesil2-SATB1-shRNA和SATB1過表達(dá)真核表達(dá)質(zhì)粒pcDNA3.1-SATB1的基礎(chǔ)上,采用CCK-8比色法評估調(diào)控SATB1的表達(dá)對RCC細(xì)胞增殖能力的影響,應(yīng)用Transwell侵襲實(shí)驗(yàn)分析RCC細(xì)胞侵襲性的變化。 結(jié)果：RCC細(xì)胞中SATB1mRNA及其蛋白表達(dá)水平顯著高于對照組細(xì)胞HK-2(P0.001),且SATB1在ACHN、A498及786-0中的表達(dá)依次增高。成功構(gòu)建pGenesil2-SATB1-shRNA和pcDNA3.1-SATB1過表達(dá)質(zhì)粒。通過Western blotting和實(shí)時(shí)定量PCR法證實(shí)：shRNA穩(wěn)轉(zhuǎn)組786-0細(xì)胞中,SATB1mRNA及其蛋白表達(dá)均顯著降低(P0.001)；而SATB1過表達(dá)組ACHN細(xì)胞中,SATB1在mRNA和蛋白水平的表達(dá)均顯著升高(P0.001)。CCK-8研究結(jié)果顯示：SATB1表達(dá)下調(diào)可顯著抑制786-0細(xì)胞的增殖能力,而SATB1過表達(dá)則可明顯促進(jìn)ACHN細(xì)胞的增殖活性,差異均具有顯著的統(tǒng)計(jì)學(xué)意義(均為P0.05)。Transwell研究表明：shRNA穩(wěn)轉(zhuǎn)組腎癌細(xì)胞786-0的侵襲能力顯著降低(P0.001),而pcDNA3.1-SATB1穩(wěn)轉(zhuǎn)組腎癌細(xì)胞ACHN的侵襲能力則顯著增強(qiáng)(P0.001)。 結(jié)論：核轉(zhuǎn)錄因子SATB1在RCC細(xì)胞中的表達(dá)顯著上調(diào)。SATB1基因過表達(dá)在腎癌細(xì)胞獲得侵襲表型的過程中起關(guān)鍵作用,對促進(jìn)腎癌侵襲、轉(zhuǎn)移具有重要意義。 目的：探討SATB1和EMT標(biāo)志基因在ccRCC組織中的表達(dá)及其相關(guān)性。 方法：分別采用免疫組織化學(xué)(IHC)染色、Western blotting及實(shí)時(shí)定量PCR法檢測89例ccRCC組織標(biāo)本中SATB1的表達(dá)情況；應(yīng)用IHC法檢測SATB2及EMT標(biāo)志基因(E-cadherin、ZEB2、vimentin和fibronectin)在ccRCC組織中的表達(dá),并通過Spearman秩次相關(guān)分析法評估其與SATB1表達(dá)之間的相關(guān)性。 結(jié)果：IHC結(jié)果顯示89例ccRCC組織中SATB1、SATB2、E-cadherin、ZEB2、vimentin和fibronectin蛋白表達(dá)陽性率分別為46.1%、42.7%、44.9%、51.7%、37.1%和49.4%。與對照組正常組織相比,腎癌組織中SATB2和E-cadherin的表達(dá)水平均明顯下降,而SATB1和ZEB2表達(dá)水平則顯著升高,且SATB1高表達(dá)與患者腫瘤浸潤深度(P0.001)、TNM分期(P=0.009)及淋巴結(jié)轉(zhuǎn)移狀態(tài)(P=0.001)之間具有顯著關(guān)聯(lián)。此外,Spearman秩次相關(guān)分析表明SATB1表達(dá)與ZEB2之間呈顯著正相關(guān)(R=0.262,P=0.013),而與SATB2、E-cadherin之間則呈明顯負(fù)相關(guān)(]R=-0.296, P=0.005; R=-0.427,P0.001)。 結(jié)論：ccRCC組織中,SATB1表達(dá)與EMT標(biāo)志基因蛋白表達(dá)密切相關(guān),將二者聯(lián)合進(jìn)行分析,對于進(jìn)一步明確SATB1與EMT過程在腎癌侵襲轉(zhuǎn)移中所起的作用以及評估腎癌患者預(yù)后均具有重要意義。
[Abstract]:Aim: to investigate the expression of nuclear transcription factor SATB1 in different renal cell lines and the effect of SATB1 gene expression on the proliferation and invasion of renal carcinoma cells. Methods: the expression of SATB1 in different RCC cell lines was detected by real-time quantitative PCR Western blotting and immunofluorescence staining. The eukaryotic expression plasmids pcDNA3.1-SATB1 of pGenesil2-SATB1-shRNA and SATB1 targeting silencing SATB1 were constructed. CCK-8 colorimetric assay was used to evaluate the effect of regulating SATB1 expression on the proliferation of RCC cells, and Transwell invasion assay was used to analyze the changes of the invasiveness of RCC cells. Results the expression of SATB1mRNA and its protein was significantly higher in the cell line than that in the control cell line HK-2P 0.001, and the expression of SATB1 was increased in turn in ACHNNAA498 and 786-0. The overexpression plasmids of pGenesil2-SATB1-shRNA and pcDNA3.1-SATB1 were successfully constructed. The stable transformation of pGenesil2-SATB1-shRNA and pcDNA3.1-SATB1 was confirmed by Western blotting and real-time quantitative PCR. In group 786-0, the expression of Tb1 mRNA and its protein decreased significantly, while the expression of mRNA and protein in ACHN cells in SATB1 overexpression group increased significantly. The results of CCK-8 study showed that down-regulation of the expression of TB1 in SATB1 could significantly inhibit the proliferation of 786-0 cells. However, overexpression of SATB1 could significantly promote the proliferation of ACHN cells. The difference was statistically significant (all P0.05).Transwell studies showed that the invasion ability of renal cancer cell line 786-0 was significantly decreased in pcDNA3.1-SATB1 stable group, while the invasion ability of ACHN was significantly enhanced in pcDNA3.1-SATB1 stable group. Conclusion: the expression of nuclear transcription factor SATB1 significantly up-regulates the expression of. SATB1 gene in RCC cells, which plays a key role in the process of invasion phenotype of renal cancer cells, and plays an important role in promoting the invasion and metastasis of renal cell carcinoma. Objective: to investigate the expression and correlation of SATB1 and EMT marker genes in ccRCC tissues. Methods: the expression of SATB1 in 89 cases of ccRCC tissues was detected by immunohistochemical staining blotting and real time quantitative PCR, and the expression of SATB2 and EMT marker genes such as SATB2 and EMT marker ZEB2vimentin and fibronectin in ccRCC tissues were detected by IHC method. The correlation between SATB1 expression and Spearman rank correlation analysis was evaluated. Results the positive rates of SATB2 and E-cadherin in 89 cases of ccRCC were 46.1% and 44.7%, respectively. Compared with the control group, the expression of SATB2 and E-cadherin in RCC were significantly decreased, while the expression of SATB1 and ZEB2 were significantly increased. Moreover, there was a significant correlation between the high expression of SATB1 and the depth of tumor invasion (P0.001, P0.009) and the metastatic status of lymph nodes (P 0.001). In addition, there was a significant positive correlation between the expression of SATB1 and ZEB2, and a significant negative correlation between the expression of SATB1 and ZEB2, but a significant negative correlation with SATB2E-cadherin (] RV -0.296, P0. 005; R- 0. 427 and P0. 0027; P 0. 0013), but negative correlation with SATB2E-cadherin (P 0. 001, P 0. 005, P 0. 005, P 0. 005, P 0. 005, P 0. 427, P 0. 0013). Conclusion the expression of SATB1 is closely related to the expression of EMT marker gene protein in the tissues of WCCRCC, and the expression of SATB1 is associated with the protein expression of the EMT marker gene. It is important to further understand the role of SATB1 and EMT in the invasion and metastasis of RCC and to evaluate the prognosis of RCC.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R737.11

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳曉霞;孫欽峰;;特殊富含AT序列結(jié)合蛋白-2的生物學(xué)特性[J];國際口腔醫(yī)學(xué)雜志;2011年03期

2 李光輝;楊定華;李湘z，

本文編號：1695965