【摘要】：目的:目前已有研究發(fā)現(xiàn)OIP5基因在人類某些癌癥中明顯高表達(dá),并已證實(shí)其與人類某些癌癥的發(fā)生發(fā)展有關(guān)。然而,OIP5基因在腎透明細(xì)胞癌(CCRCC)中的表達(dá)情況和臨床意義仍然不明確。因此,本研究旨在探討OIP5基因在腎癌組織及腎癌細(xì)胞系中的表達(dá)情況及作用,同時(shí)研究OIP5特異性的siRNA與三氧化二砷聯(lián)合應(yīng)用時(shí)增強(qiáng)三氧化二砷的抗腫瘤作用,為尋找腎癌的標(biāo)記物以及腎癌的治療方法提供理論基礎(chǔ)。方法:我們應(yīng)用RT-PCR檢測(cè)了OIP5在腎癌組織及癌旁正常組織中的表達(dá)情況。在103例石蠟包埋的腎癌組織中,我們應(yīng)用免疫組化方法檢測(cè)了OIP5的表達(dá)情況,并分析了OIP5與這103例患者臨床病理特征及預(yù)后之間的關(guān)系。除此之外,我們利用轉(zhuǎn)染siRNA的方法下調(diào)786-O和Caki-2細(xì)胞中OIP5的表達(dá),用CCK8和克隆形成實(shí)驗(yàn)檢測(cè)細(xì)胞活力和增殖的情況。最后,我們用CCK8檢測(cè)了OIP5特異性的siRNA聯(lián)合三氧化二砷對(duì)細(xì)胞增殖活力的影響。結(jié)果:在本研究中,我們發(fā)現(xiàn)OIP5基因在腎透明細(xì)胞癌組織和腎透明細(xì)胞癌細(xì)胞系中的表達(dá)均顯著上調(diào)。免疫組化結(jié)果顯示,腎透明細(xì)胞癌組織中OIP5的表達(dá)水平高于腎正常組織。進(jìn)一步的統(tǒng)計(jì)分析表明OIP5的上調(diào)與Fuhrman分級(jí)(P=0.02),T分期(P=0.015),N分期(P=0.018)和臨床分期(P=0.035)呈正相關(guān)。此外,OIP5高表達(dá)的腎癌患者比OIP5低表達(dá)患者生存時(shí)間縮短,差異具有統(tǒng)計(jì)學(xué)意義(P=0.001)。而且,本研究通過多因素分析表明OIP5的表達(dá)是腎透明細(xì)胞癌患者總生存期的一個(gè)獨(dú)立預(yù)后指標(biāo)(P=0.008)。CCK-8和克隆實(shí)驗(yàn)結(jié)果顯示,利用siRNA將OIP5沉默后能顯著抑制腎癌細(xì)胞的活力和增殖。另一方面,CCK-8結(jié)果顯示,OIP5特異性的siRNA與三氧化二砷聯(lián)用,能更顯著的抑制腎癌細(xì)胞的活力。結(jié)論:本研究首次表明OIP5基因在腎透明細(xì)胞癌組織和細(xì)胞系中呈高表達(dá)。OIP5的下調(diào)有效地抑制了腎透明細(xì)胞癌細(xì)胞的增殖。此外,我們的研究表明OIP5基因的表達(dá)是腎透明細(xì)胞癌患者預(yù)后的一個(gè)獨(dú)立指標(biāo)。最后OIP5基因的下調(diào)可以增強(qiáng)三氧化二砷的抗腫瘤作用,減少三氧化二砷的用量。
[Abstract]:Objective: it has been found that OIP5 gene is significantly overexpressed in some human cancers and has been proved to be related to the occurrence and development of some human cancers. However, the expression and clinical significance of OIP5 gene in (CCRCC) of renal clear cell carcinoma are still unclear. Therefore, the purpose of this study was to investigate the expression and role of OIP5 gene in renal cell carcinoma tissues and cell lines, and to study the anti-tumor effect of OIP5 specific siRNA combined with arsenic trioxide. To find the marker of renal cell carcinoma and the treatment of renal cell carcinoma to provide a theoretical basis. Methods: RT-PCR was used to detect the expression of OIP5 in renal carcinoma and adjacent normal tissues. The expression of OIP5 was detected by immunohistochemical method in 103 cases of paraffin embedded renal cell carcinoma, and the relationship between OIP5 and clinicopathological features and prognosis was analyzed. In addition, we down-regulated the expression of OIP5 in 786-O and Caki-2 cells by siRNA transfection, and detected cell viability and proliferation by CCK8 and clone formation assay. Finally, CCK8 was used to detect the effect of OIP5 specific siRNA combined with arsenic trioxide on cell proliferation. Results: in this study, we found that the expression of OIP5 gene was significantly up-regulated in both renal clear cell carcinoma tissues and renal clear cell carcinoma cell lines. Immunohistochemical results showed that the expression of OIP5 in renal clear cell carcinoma was higher than that in normal renal tissue. Further statistical analysis showed that the upregulation of OIP5 was positively correlated with Fuhrman grade (P0. 02) / T stage (P0. 015) and clinical stage (P0. 018) and clinical stage (P0. 035). In addition, the survival time of patients with high expression of OIP5 was shorter than that of patients with low expression of OIP5 (P0. 001). Furthermore, multivariate analysis showed that the expression of OIP5 was an independent prognostic marker (P0. 008). CCK-8 and clone assay showed that siRNA silencing OIP5 could significantly inhibit the activity and proliferation of renal cancer cells. On the other hand, CCK-8 showed that OIP5 specific siRNA combined with arsenic trioxide could significantly inhibit the activity of renal cancer cells. Conclusion: this study shows for the first time that the down-regulation of overexpression of OIP5 gene in renal clear cell carcinoma tissues and cell lines can effectively inhibit the proliferation of renal clear cell carcinoma cells. In addition, our study suggests that OIP5 gene expression is an independent predictor of prognosis in patients with renal clear cell carcinoma. Finally, the down-regulation of OIP5 gene can enhance the antitumor effect of arsenic trioxide and reduce the dosage of arsenic trioxide.
【學(xué)位授予單位】：哈爾濱醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.11

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