【摘要】 目的:探讨LOC100130476在肾细胞癌(renal cell carcinoma, RCC)中的表达及启动子区甲基化状态。方法:应用RT-qPCR和甲基化特异性PCR(methylation-specific PCR,MSP)检测RCC中LOC100130476的表达及启动子区甲基化状态,并分析与临床病理资料的关系;DNA甲基转移酶抑制剂(5-Aza-dC)干预后的肾癌细胞系中,采用RT-qPCR检测LOC100130476的表达及甲基化状态。结果:RT-qPCR显示,LOC100130476在RCC组织的表达明显低于正常组织(0.593±0.306 vs 1.000±0.018,t=-11.143,P<0.001),并与淋巴结转移(t=2.774,P=0.007)及临床分期(F=5.436,P=0.002)相关。LOC100130476在3株肾癌细胞系(A498、ACHN、786-O)的相对表达量均低于对照组(HK-2)(F=107.068,P<0.001)。5-Aza-dC处理后,LOC100130476的表达均增高。MSP显示,RCC组织的甲基化率明显高于正常组织(χ~2=33.273,P<0.01),并与淋巴结转移有关(χ~2=4.250,P=0.039);3株肾癌细胞系经5-Aza-dC处理后,A498、786-O中LOC100130476甲基化消失,ACHN中LOC100130476甲基化降低,3种细胞中LOC100130476非甲基化增高。结论:LOC100130476在RCC中低表达,并呈高甲基化状态,其高甲基化可能是肾癌发生发展的分子机制之一。更多还原

【Abstract】 Objective: To investigate the expression of LOC100130476 in renal cell carcinoma(RCC) and the methylation status of the promoter region. Methods: We used RT-qPCR and methylation-specific PCR(MSP) to detect the expression of LOC100130476 in RCC and the methylation status of the promoter region, and analyzed the relationship between LOC100130476 expression and clinicopathological characteristics. RT-qPCR was used to detect the expression and methylation status of LOC100130476 in renal cancer cell lines intervened with DNA methyltransferase inhibitor(5-Aza-dC). Results: RT-qPCR showed that the expression of LOC100130476 in RCC tissue was significantly lower than that in normal tissue(0.593±0.306 vs 1.000±0.018, t=-11.143, P<0.001), and LOC100130476 expression was related to lymph node metastasis(t=2.774, P=0.007) and clinical stage(F=5.436, P=0.002). The relative expression levels of LOC100130476 in three RCC cell lines(A498, ACHN and 786-O) were lower than that in the control group(HK-2)(F=107.068, P<0.001). The expression levels of LOC10013 0476 in A498, ACHN and 786-O increased after being treated with 5-Aza-dC. MSP showed that the methylation rate in RCC tissue was significantly higher than that in normal tissue(χ~2=33.273, P<0.01), and it was related to lymph node metastasis(χ~2=4.250, P=0.039); after being treated with 5-Aza-dC, the methylation of A498 and 786-O disappeared, that of ACHN decreased, and the unmethylation of A498, ACHN and 786-O increased. Conclusion: LOC100130476 is lowly expressed and hypermethylated in RCC. Its hypermethylation may be one of the molecular mechanisms of the occurrence and development of RCC.更多还原