肾细胞癌中Wif-1基因启动子区甲基化状态与mRNA表达的研究
发布时间：2018-07-25 14:40
【摘要】：目的：Wif-1（Wnt inhibitory factor-1，Wif-1）是Wnt信号转导通路的一个重要抑制因子，国内外大量实验发现在众多恶性肿瘤中存在Wif-1的表达异常或启动子区的高甲基化，，提示启动子区的高甲基化可能是肿瘤发生机制之一。但是该基因在肾细胞癌（Renal Cell Carcinoma，RCC）中的研究较少，本实验通过应用甲基化特异性聚合酶链反应（Methylation-Specific PCR，MSP）和逆转录-聚合酶链反应（ReverseTranscriptase-PCR，RT-PCR）探讨RCC组织及正常肾组织中Wif-1基因启动子甲基化状态与mRNA表达水平，并研究它们与临床资料之间的关系，为RCC的发病机制、临床诊断及治疗提供一定的理论依据。 方法： 1应用甲基化特异性聚合酶链反应（Methylation-Specific PCR，MSP）检测56例RCC组织及正常肾组织中Wif-1基因甲基化状态，并分析该基因甲基化在RCC发生发展中的作用以及与临床资料之间的关系。 2应用逆转录-聚合酶链反应（Reverse Transcriptase-PCR，RT-PCR）技术检测Wif-1基因mRNA在56例RCC组织及正常肾组织中的相对表达量，并分析在RCC组织中Wif-1基因mRNA相对表达量与临床资料之间的关系。 3应用统计软件SPSS17.0对数据进行统计学分析。 结果： 156例RCC组织和56例正常肾组织中Wif-1基因启动子的甲基化情况以及RCC组织中甲基化与临床资料之间的研究。 Wif-1基因在RCC组织甲基化率分别是46.4％（26/56）明显高于正常肾组织的甲基化率3.6％（2/56），P0.05，差异具有统计学意义。但是Wif-1的甲基化率与性别、年龄、临床分期及肿瘤大小等临床资料无明显关系。 256例RCC组织和56例正常肾组织中Wif-1基因mRNA相对表达量情况，以及RCC组织中Wif-1基因mRNA相对表达量与临床资料之间的关系。 Wif-1基因mRNA在RCC组织中的相对表达量0.65±0.17低于正常肾组织mRNA的相对表达量0.77±0.06，经统计学分析，P0.05，两组差异有意义。但是Wif-1基因mRNA相对表达量与年龄、性别、肿瘤大小等临床资料无统计学意义。 3在RCC组织中，Wif-1基因mRNA的相对表达量在甲基化阳性病例中为0.68±0.17高于甲基化阴性病例中mRNA的相对表达量0.63±0.17，P0.05，但是差异无统计学意义。 结论： 1RCC组织中Wif-1基因的甲基化率46.4％（26/56）明显高于正常肾组织的甲基化率3.6％（2/56），经统计学分析，P0.05，两组甲基化率的差异有意义，该研究提示Wif-1基因启动子区的高甲基化在RCC的发生中是一个常见事件，并且可能参与RCC的发生。 2Wif-1基因mRNA在RCC组织中的相对表达量0.65±0.17低于正常肾组织中的相对表达量0.77±0.06，经统计学分析，P0.05，提示Wif-1基因mRNA表达的降低可能参与RCC的发生发展过程。 3在RCC组织中，Wif-1基因mRNA的相对表达量在甲基化阳性病例中为0.68±0.17高于甲基化阴性病例中mRNA的相对表达量0.63±0.17，P0.05，但是差异无统计学意义，提示Wif-1基因启动子高甲基化可能只是RCC中的一个常见事件，而不会导致其mRNA表达的降低，其表达降低的具体原因有待进一步研究。
[Abstract]:Objective: Wif-1 (Wnt inhibitory factor-1, Wif-1) is an important inhibitory factor in the Wnt signal transduction pathway. A large number of experiments at home and abroad have found that the expression of Wif-1 is abnormal or the hypermethylation of the promoter region in many malignant tumors, suggesting that hypermethylation of the promoter region may be one of the mechanisms of the tumor, but the gene is in the renal cell. There are few studies in Renal Cell Carcinoma (RCC). The methylation status and mRNA expression level of Wif-1 gene promoter in RCC tissues and normal renal tissues were investigated by using the methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP) and reverse transcription polymerase chain reaction (ReverseTranscriptase-PCR, RT-PCR). The relationship between them and clinical data provides a theoretical basis for the pathogenesis, clinical diagnosis and treatment of RCC.
Method:
1 methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP) was used to detect the methylation status of Wif-1 gene in 56 cases of RCC and normal renal tissues, and the role of methylation in the development of RCC and the relationship with clinical data were analyzed.
2 the relative expression of Wif-1 gene mRNA in 56 cases of RCC tissue and normal renal tissue was detected by reverse transcription polymerase chain reaction (Reverse Transcriptase-PCR), and the relationship between the relative expression of mRNA and the clinical data of the Wif-1 gene in the RCC tissues was analyzed.
3 statistical analysis was performed using statistical software SPSS17.0.
Result:
Methylation of Wif-1 gene promoter and methylation in RCC tissues and clinical data in 156 cases of RCC and 56 normal renal tissues were studied.
The methylation rate of Wif-1 gene in RCC tissues was 46.4% (26/56), respectively, which was significantly higher than that of normal renal tissue by 3.6% (2/56) and P0.05. The difference was statistically significant, but the methylation rate of Wif-1 was not significantly related to the clinical data such as sex, age, clinical stage and tumor size.
The relative expression of Wif-1 gene mRNA in 256 cases of RCC and 56 normal renal tissues, as well as the relationship between the relative expression of the Wif-1 gene mRNA in the RCC tissue and the clinical data.
The relative expression of Wif-1 gene mRNA in RCC tissues was 0.65 + 0.17 lower than that of normal renal tissue mRNA. The relative expression of mRNA was 0.77 + 0.06. Statistically, P0.05 and two groups were significant. However, the relative expression of Wif-1 gene mRNA has no significance in clinical data such as age, sex, tumor size and other clinical data.
3 in the RCC tissue, the relative expression of the Wif-1 gene mRNA in the methylation positive cases was 0.68 + 0.17 higher than that in the negative methylation cases, the relative expression of mRNA was 0.63 + 0.17, P0.05, but the difference was not statistically significant.
Conclusion:
The methylation rate of Wif-1 gene in 1RCC tissues 46.4% (26/56) is significantly higher than that of normal renal tissue by 3.6% (2/56). Statistically, P0.05 and two groups of methylation rates are significant. This study suggests that hypermethylation of the Wif-1 gene promoter region is a common event in the occurrence of RCC and may be involved in the occurrence of RCC.
The relative expression of 2Wif-1 gene mRNA in RCC tissues is 0.65 + 0.17 lower than that of normal renal tissue, which is 0.77 + 0.06, P0.05, suggesting that the decrease of mRNA expression of Wif-1 gene may be involved in the development of RCC.
3 in RCC tissues, the relative expression of the Wif-1 gene mRNA was 0.68 + 0.17 in the positive methylation cases, and the relative expression of mRNA in the negative methylation cases was 0.63 + 0.17, P0.05, but the difference was not statistically significant. It suggested that the hypermethylation of the Wif-1 gene promoter may be a common event in RCC, without causing its mRNA expression. The specific reasons for the decrease of expression should be further studied.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.11
本文编号：2144138
[Abstract]:Objective: Wif-1 (Wnt inhibitory factor-1, Wif-1) is an important inhibitory factor in the Wnt signal transduction pathway. A large number of experiments at home and abroad have found that the expression of Wif-1 is abnormal or the hypermethylation of the promoter region in many malignant tumors, suggesting that hypermethylation of the promoter region may be one of the mechanisms of the tumor, but the gene is in the renal cell. There are few studies in Renal Cell Carcinoma (RCC). The methylation status and mRNA expression level of Wif-1 gene promoter in RCC tissues and normal renal tissues were investigated by using the methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP) and reverse transcription polymerase chain reaction (ReverseTranscriptase-PCR, RT-PCR). The relationship between them and clinical data provides a theoretical basis for the pathogenesis, clinical diagnosis and treatment of RCC.
Method:
1 methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP) was used to detect the methylation status of Wif-1 gene in 56 cases of RCC and normal renal tissues, and the role of methylation in the development of RCC and the relationship with clinical data were analyzed.
2 the relative expression of Wif-1 gene mRNA in 56 cases of RCC tissue and normal renal tissue was detected by reverse transcription polymerase chain reaction (Reverse Transcriptase-PCR), and the relationship between the relative expression of mRNA and the clinical data of the Wif-1 gene in the RCC tissues was analyzed.
3 statistical analysis was performed using statistical software SPSS17.0.
Result:
Methylation of Wif-1 gene promoter and methylation in RCC tissues and clinical data in 156 cases of RCC and 56 normal renal tissues were studied.
The methylation rate of Wif-1 gene in RCC tissues was 46.4% (26/56), respectively, which was significantly higher than that of normal renal tissue by 3.6% (2/56) and P0.05. The difference was statistically significant, but the methylation rate of Wif-1 was not significantly related to the clinical data such as sex, age, clinical stage and tumor size.
The relative expression of Wif-1 gene mRNA in 256 cases of RCC and 56 normal renal tissues, as well as the relationship between the relative expression of the Wif-1 gene mRNA in the RCC tissue and the clinical data.
The relative expression of Wif-1 gene mRNA in RCC tissues was 0.65 + 0.17 lower than that of normal renal tissue mRNA. The relative expression of mRNA was 0.77 + 0.06. Statistically, P0.05 and two groups were significant. However, the relative expression of Wif-1 gene mRNA has no significance in clinical data such as age, sex, tumor size and other clinical data.
3 in the RCC tissue, the relative expression of the Wif-1 gene mRNA in the methylation positive cases was 0.68 + 0.17 higher than that in the negative methylation cases, the relative expression of mRNA was 0.63 + 0.17, P0.05, but the difference was not statistically significant.
Conclusion:
The methylation rate of Wif-1 gene in 1RCC tissues 46.4% (26/56) is significantly higher than that of normal renal tissue by 3.6% (2/56). Statistically, P0.05 and two groups of methylation rates are significant. This study suggests that hypermethylation of the Wif-1 gene promoter region is a common event in the occurrence of RCC and may be involved in the occurrence of RCC.
The relative expression of 2Wif-1 gene mRNA in RCC tissues is 0.65 + 0.17 lower than that of normal renal tissue, which is 0.77 + 0.06, P0.05, suggesting that the decrease of mRNA expression of Wif-1 gene may be involved in the development of RCC.
3 in RCC tissues, the relative expression of the Wif-1 gene mRNA was 0.68 + 0.17 in the positive methylation cases, and the relative expression of mRNA in the negative methylation cases was 0.63 + 0.17, P0.05, but the difference was not statistically significant. It suggested that the hypermethylation of the Wif-1 gene promoter may be a common event in RCC, without causing its mRNA expression. The specific reasons for the decrease of expression should be further studied.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.11
【参考文献】
相关期刊论文 前1条
1 ;Promoter methylation and mRNA expression of DKK-3 and WIF-1 in hepatocellular carcinoma[J];World Journal of Gastroenterology;2009年21期
本文编号：2144138
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