腎細(xì)胞癌中Wif-1基因啟動(dòng)子區(qū)甲基化狀態(tài)與mRNA表達(dá)的研究
發(fā)布時(shí)間：2018-07-25 14:40
【摘要】：目的：Wif-1（Wnt inhibitory factor-1，Wif-1）是Wnt信號(hào)轉(zhuǎn)導(dǎo)通路的一個(gè)重要抑制因子，國內(nèi)外大量實(shí)驗(yàn)發(fā)現(xiàn)在眾多惡性腫瘤中存在Wif-1的表達(dá)異�；騿�(dòng)子區(qū)的高甲基化，，提示啟動(dòng)子區(qū)的高甲基化可能是腫瘤發(fā)生機(jī)制之一。但是該基因在腎細(xì)胞癌（Renal Cell Carcinoma，RCC）中的研究較少，本實(shí)驗(yàn)通過應(yīng)用甲基化特異性聚合酶鏈反應(yīng)（Methylation-Specific PCR，MSP）和逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)（ReverseTranscriptase-PCR，RT-PCR）探討RCC組織及正常腎組織中Wif-1基因啟動(dòng)子甲基化狀態(tài)與mRNA表達(dá)水平，并研究它們與臨床資料之間的關(guān)系，為RCC的發(fā)病機(jī)制、臨床診斷及治療提供一定的理論依據(jù)。 方法： 1應(yīng)用甲基化特異性聚合酶鏈反應(yīng)（Methylation-Specific PCR，MSP）檢測56例RCC組織及正常腎組織中Wif-1基因甲基化狀態(tài)，并分析該基因甲基化在RCC發(fā)生發(fā)展中的作用以及與臨床資料之間的關(guān)系。 2應(yīng)用逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)（Reverse Transcriptase-PCR，RT-PCR）技術(shù)檢測Wif-1基因mRNA在56例RCC組織及正常腎組織中的相對表達(dá)量，并分析在RCC組織中Wif-1基因mRNA相對表達(dá)量與臨床資料之間的關(guān)系。 3應(yīng)用統(tǒng)計(jì)軟件SPSS17.0對數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果： 156例RCC組織和56例正常腎組織中Wif-1基因啟動(dòng)子的甲基化情況以及RCC組織中甲基化與臨床資料之間的研究。 Wif-1基因在RCC組織甲基化率分別是46.4％（26/56）明顯高于正常腎組織的甲基化率3.6％（2/56），P0.05，差異具有統(tǒng)計(jì)學(xué)意義。但是Wif-1的甲基化率與性別、年齡、臨床分期及腫瘤大小等臨床資料無明顯關(guān)系。 256例RCC組織和56例正常腎組織中Wif-1基因mRNA相對表達(dá)量情況，以及RCC組織中Wif-1基因mRNA相對表達(dá)量與臨床資料之間的關(guān)系。 Wif-1基因mRNA在RCC組織中的相對表達(dá)量0.65±0.17低于正常腎組織mRNA的相對表達(dá)量0.77±0.06，經(jīng)統(tǒng)計(jì)學(xué)分析，P0.05，兩組差異有意義。但是Wif-1基因mRNA相對表達(dá)量與年齡、性別、腫瘤大小等臨床資料無統(tǒng)計(jì)學(xué)意義。 3在RCC組織中，Wif-1基因mRNA的相對表達(dá)量在甲基化陽性病例中為0.68±0.17高于甲基化陰性病例中mRNA的相對表達(dá)量0.63±0.17，P0.05，但是差異無統(tǒng)計(jì)學(xué)意義。 結(jié)論： 1RCC組織中Wif-1基因的甲基化率46.4％（26/56）明顯高于正常腎組織的甲基化率3.6％（2/56），經(jīng)統(tǒng)計(jì)學(xué)分析，P0.05，兩組甲基化率的差異有意義，該研究提示W(wǎng)if-1基因啟動(dòng)子區(qū)的高甲基化在RCC的發(fā)生中是一個(gè)常見事件，并且可能參與RCC的發(fā)生。 2Wif-1基因mRNA在RCC組織中的相對表達(dá)量0.65±0.17低于正常腎組織中的相對表達(dá)量0.77±0.06，經(jīng)統(tǒng)計(jì)學(xué)分析，P0.05，提示W(wǎng)if-1基因mRNA表達(dá)的降低可能參與RCC的發(fā)生發(fā)展過程。 3在RCC組織中，Wif-1基因mRNA的相對表達(dá)量在甲基化陽性病例中為0.68±0.17高于甲基化陰性病例中mRNA的相對表達(dá)量0.63±0.17，P0.05，但是差異無統(tǒng)計(jì)學(xué)意義，提示W(wǎng)if-1基因啟動(dòng)子高甲基化可能只是RCC中的一個(gè)常見事件，而不會(huì)導(dǎo)致其mRNA表達(dá)的降低，其表達(dá)降低的具體原因有待進(jìn)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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.11
本文編號(hào)：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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.11
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 ;Promoter methylation and mRNA expression of DKK-3 and WIF-1 in hepatocellular carcinoma[J];World Journal of Gastroenterology;2009年21期
本文編號(hào)：2144138
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