肾细胞癌SOX7基因启动子区甲基化状态及mRNA表达的研究

发布时间：2018-02-12 08:59

本文关键词： 肾细胞癌 SOX7 启动子甲基化基因表达　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:肾细胞癌(Renal Cell Carcinoma,RCC)是起源于肾实质泌尿小管上皮系统的恶性肿瘤,简称肾癌,其发病率约为4/10万,严重威胁着我国人群的健康。其发病原因尚不明确,可能与遗传、肥胖、吸烟、高血压以及抗高血压治疗有关。研究发现表观遗传学改变可以影响基因的表达,而这可能与肿瘤的发生、发展有着密切的关系。目前,研究较深入的是表观遗传学中的基因甲基化与肿瘤间的关系。SOX7属于SOXF亚族,参与调节多种发育过程,如脉管的形成、血液的形成、肌肉的发育等。近年来有研究结果表明在肺癌、乳腺癌、肝癌等恶性肿瘤中存在SOX7基因启动子甲基化及SOX7m RNA表达异常,这可能是导致肿瘤发生以及发展的原因之一。然而目前尚无SOX7在RCC中相关报道。本研究通过特异性PCR及RT-PCR技术来分析SOX7基因异常甲基化与转录表达之间的关系,并通过统计学分析来研究SOX7基因异常甲基化与患者临床数据之间的关系。初步探讨SOX7基因异常甲基化在RCC中的作用。方法:1收集河北医科大学第四医院肾癌组织及正常组织标本58例。采用反转录-聚合酶链反应(Reverse Transcriptase-PCR,RT-PCR)来检测58例RCC患者癌组织及正常组织中SOX7基因表达情况。2采用甲基化特异性PCR(Methylation Specific PCR,MSP)分析58例肾癌组织及正常肾组织中SOX7基因启动子区的甲基化状态。3使用统计学软件SPSS20.0对实验数据进行统计学分析。结果:1 SOX7在RCC组织和正常组织中存在完全甲基化、不完全甲基化、完全非甲基化三种不同的甲基化状态。2 SOX7基因在RCC组织中甲基化阳性率为36.2%(21/58)高于正常组织标本中的甲基化率阳性率10.3%(6/58),二者差异具有统计学意义(χ2=10.86 P=0.001);58例肾癌组织中,SOX7基因甲基化率在年龄(χ2=0.015 P=0.559)、性别(χ2=0.023P=0.547)、肿瘤大小(χ2=0.037 P=0.547)、Fuhrman核分级(χ2=0.514 P=0.773)四个方面没有统计学差异(P值均大于0.05);在临床分期(Ⅰ、Ⅱ期与Ⅲ、Ⅳ期)(P=0.035)方面存在统计学差异(P值小于0.05)。3肾癌组织中SOX7基因m RNA的相对表达量为0.35±0.06,低于正常组织标本中的相对表达量0.62±0.07,经过统计学分析,差异具有统计学意义(t=-22.679,P=0.001)。肾癌组织中,SOX7基因的相对表达量在年龄(t=1.359 P=0.182)、性别(t=-0.490 P=0.626)、肿瘤大小(t=-1.347 P=0.183)三方面没有统计学差异(P值均大于0.05);在临床分期(Ⅰ、Ⅱ期与Ⅲ、Ⅳ期)(t=-2.254,P=0.028)方面存在统计学差异(P值小于0.05)。4 58例患者在肾癌组织标本方面,甲基化阳性患者SOX7基因m RNA的相对表达量为0.41±0.05,甲基化阴性患者SOX7基因m RNA的相对表达量为0.63±0.07,并且其差异存在统计学意义(t=-12.506,P=0.000)。结论:1肾癌组织中SOX7基因启动子区高甲基化是频发事件,并且肾癌组织中SOX7基因的甲基化率明显高于正常肾组织,提示SOX7基因的异常甲基化可能是肾癌发生的主要机制之一。2肾癌组织中SOX7基因m RNA的相对表达量较正常组织低,提示SOX7基因在肾癌的发生发展可能起着抑癌基因的作用。3肾癌组织中甲基化阳性组SOX7m RNA的相对表达较非甲基化组低,提示SOX7基因甲基化有可能是导致其表达沉默的重要原因。
[Abstract]:Objective: renal cell carcinoma (Renal Cell, Carcinoma, RCC) is originated in the renal tubular epithelium of urinary system cancer, RCC, its incidence rate is about 4/10 million, a serious threat to our health. Its pathogenesis is not clear, may be related to heredity, obesity, smoking, hypertension and anti the treatment of hypertension. The study found that epigenetic changes can affect gene expression, which might be related to tumorigenesis is closely related to development. At present, in-depth study is the apparent relationship between.SOX7 gene methylation and tumor genetics in the SOXF subfamily belongs to, participate in the regulation of diverse developmental processes, such as vascular the formation of blood formation, muscle development. In recent years, the results of the study show that in lung cancer, breast cancer, SOX7 gene promoter methylation and SOX7m expression of RNA in abnormal liver cancer and other malignant tumors, which may lead to tumor One of the reasons for occurrence and development. However, there is no SOX7 in RCC reports. Through the study of specific PCR and RT-PCR technique to analyze the relationship between the expression of SOX7 gene methylation and transcription, and through statistical analysis to study the relationship between abnormal methylation of SOX7 gene in patients with clinical data. A preliminary study of SOX7 gene abnormality methylation in RCC. Methods: 1 collected from the fourth hospital of Hebei Medical University renal cell carcinoma and normal tissues of 58 cases. By using reverse transcription polymerase chain reaction (Reverse Transcriptase-PCR RT-PCR) to SOX7 were measured in 58 cases of RCC cancer tissue and normal tissue, the expression of.2 gene by methylation specific PCR (Methylation Specific PCR MSP, SOX7) analysis of 58 cases of renal cell carcinoma and normal renal tissue gene promoter methylation status of.3 using statistical software SPSS20.0 on experimental data Statistical analysis was performed. Results: 1 SOX7 methylation in RCC and normal tissue, incomplete methylation, completely unmethylated in three different methylation status of.2 gene methylation positive rate of SOX7 in RCC tissues was 36.2% (21/58) higher than the methylation positive rate of 10.3% normal tissues (6/58), two the difference was statistically significant (2=10.86 P=0.001); 58 cases of renal cell carcinoma, SOX7 gene methylation rate in age (2=0.015 P=0.559), gender (2=0.023P=0.547), tumor size (2=0.037 P=0.547), Fuhrman nuclear grade (2=0.514 P=0.773) had no statistically significant differences between the four aspects (the P values were greater than 0.05); the clinical stages (I, II and III, IV) (P=0.035) have significant differences (P value less than 0.05) the relative expression of SOX7 gene m.3 in renal cell carcinoma RNA was 0.35 + 0.06, lower than the relative expression of normal tissues 0.62 + 0.07, through statistical analysis, the difference was statistically significant (t=-22.679, P=0.001). Renal cell carcinoma, the expression of SOX7 (t=1.359 P=0.182) in the age, gender, tumor size (t=-0.490 P=0.626) (t=-1.347 P=0.183) three no statistical differences (P values were greater than 0.05) in the clinical stage; (I, II and III, IV) (t=-2.254, P=0.028) have significant differences (P value less than 0.05) of 58 cases of.4 patients in renal cell carcinoma tissues, the relative expression of methylation of SOX7 gene in patients with m the amount of RNA was 0.41 + 0.05, the relative expression of methylation of SOX7 gene in patients with m negative RNA the amount is 0.63 + 0.07, and the difference was statistically significant (t=-12.506, P=0.000). Conclusion: SOX7 1 renal cell carcinoma gene promoter hypermethylation is a frequent event, and the methylation rate of SOX7 gene in renal cell carcinoma was significantly higher than that in normal renal tissues, suggesting that SOX7 Aberrant methylation gene may be one of the main mechanisms of SOX7 relative expression of renal cell carcinoma of renal cell carcinoma.2 gene m RNA was lower than normal tissue, suggesting that SOX7 gene may play a role in.3 methylation of tumor suppressor gene in renal cell carcinoma of the positive group SOX7m RNA relative expression than unmethylated group on the occurrence and development of renal cell carcinoma that suggests that methylation of SOX7 gene may be an important reason lead to the expression of silence.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.11

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