OIP5基因在肾癌中的表达及其与三氧化二砷的作用关系

发布时间：2018-08-07 18:25

【摘要】：目的:目前已有研究发现OIP5基因在人类某些癌症中明显高表达,并已证实其与人类某些癌症的发生发展有关。然而,OIP5基因在肾透明细胞癌(CCRCC)中的表达情况和临床意义仍然不明确。因此,本研究旨在探讨OIP5基因在肾癌组织及肾癌细胞系中的表达情况及作用,同时研究OIP5特异性的siRNA与三氧化二砷联合应用时增强三氧化二砷的抗肿瘤作用,为寻找肾癌的标记物以及肾癌的治疗方法提供理论基础。方法:我们应用RT-PCR检测了OIP5在肾癌组织及癌旁正常组织中的表达情况。在103例石蜡包埋的肾癌组织中,我们应用免疫组化方法检测了OIP5的表达情况,并分析了OIP5与这103例患者临床病理特征及预后之间的关系。除此之外,我们利用转染siRNA的方法下调786-O和Caki-2细胞中OIP5的表达,用CCK8和克隆形成实验检测细胞活力和增殖的情况。最后,我们用CCK8检测了OIP5特异性的siRNA联合三氧化二砷对细胞增殖活力的影响。结果:在本研究中,我们发现OIP5基因在肾透明细胞癌组织和肾透明细胞癌细胞系中的表达均显著上调。免疫组化结果显示,肾透明细胞癌组织中OIP5的表达水平高于肾正常组织。进一步的统计分析表明OIP5的上调与Fuhrman分级(P=0.02),T分期(P=0.015),N分期(P=0.018)和临床分期(P=0.035)呈正相关。此外,OIP5高表达的肾癌患者比OIP5低表达患者生存时间缩短,差异具有统计学意义(P=0.001)。而且,本研究通过多因素分析表明OIP5的表达是肾透明细胞癌患者总生存期的一个独立预后指标(P=0.008)。CCK-8和克隆实验结果显示,利用siRNA将OIP5沉默后能显著抑制肾癌细胞的活力和增殖。另一方面,CCK-8结果显示,OIP5特异性的siRNA与三氧化二砷联用,能更显著的抑制肾癌细胞的活力。结论:本研究首次表明OIP5基因在肾透明细胞癌组织和细胞系中呈高表达。OIP5的下调有效地抑制了肾透明细胞癌细胞的增殖。此外,我们的研究表明OIP5基因的表达是肾透明细胞癌患者预后的一个独立指标。最后OIP5基因的下调可以增强三氧化二砷的抗肿瘤作用,减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.
【学位授予单位】：哈尔滨医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.11

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