PTTG1基因启动子区CpG岛的甲基化状态在散发性结直肠癌中的检测及其临床意义

发布时间：2018-03-03 13:53

本文选题：PTTG1　切入点：结直肠癌　出处：《福建医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：【目的】亚磷酸盐测序法(BSP)检测40例散发型性结直肠癌患者肿瘤组织及瘤旁相对正常的肠粘膜组织PTTG1基因启动子区Cp G岛甲基化率,蛋白质印迹法(Western Blot)及实时荧光定量PCR(q RT-PCR)检测PTTG1基因的表达情况,并进一步研究其与病理及临床特征之间的关系。【方法】40例散发性结直肠癌患者癌组织及癌旁相对正常的肠粘膜组织:亚磷酸盐测序法检测PTTG1基因启动子区Cp G岛(-1~-400bp区域)的甲基化率;蛋白质印记法(WB)和实时荧光定量PCR(q RT-PCR)技术分别检测PTTG1蛋白和m RNA在癌组织及癌旁组织表达;分析其PTTG1基因的表达与病理学特征及临床关系。【结果】1.亚硫酸盐测序法(BSP)检测40例结直肠癌患者的癌组织及癌旁相对正常肠粘膜组织PTTG1基因启动子Cp G岛(-1~-400bp区域)范围内共有32个可甲基化位点。其中肿瘤组织组甲基化率19.05±4.45,癌旁组为68.07±3.71,差异有统计意(P=0.032,P0.05)。结直肠癌组织PTTG1基因启动子区Cp G岛的甲基化率低于癌旁组织,癌组织的甲基化率与肿瘤分化程度、浸润深度相关,与淋巴结转移、年龄、性别等因素无关;2.免疫印记实验法(WB)检测40例结直肠癌患者的癌组织及癌旁相对正常肠粘膜组织中PTTG1蛋白表达量分别为2.79±0.23和1.00±0.17,差异有统计学意义(P0.05)。直肠癌癌组织PTTG1蛋白高表达与癌旁组织,PTTG1蛋白在直肠癌组织的高表达与肿瘤的浸润深度、分化程度相关,与患者的年龄、性别以及肿瘤大小和淋巴转移等无关;3.实时荧光定量PCR(q RT-PCR)检测40例结直肠癌患者的癌组织及癌旁相对正常肠粘膜组织中PTTG1 m RNA表达量的2-ΔΔCT值分别为18.38±2.14和9.37±1.49,差异有统计学意义(P=0.028,P0.05),显示结直肠癌组织中PTTG1 m RNA表达水平较癌旁组织升高。【结论】1.PTTG1启动子区内Cp G岛(-1~-400bp区域)癌组织的甲基化率低于癌旁组织;2.PTTG1基因在散发性结直肠癌组织中高表达,其与分化程度、浸润深度相关。可能为结直肠癌发病机制、诊断、治疗、预防提供新的思路。
[Abstract]:[objective] to detect the methylation rate of CpG island in the promoter region of PTTG1 gene in 40 patients with sporadic colorectal cancer and adjacent normal intestinal mucosa by phosphite sequencing. Western blotting and real-time fluorescence quantitative PCR(q RT-PCR were used to detect the expression of PTTG1 gene. [methods] 40 patients with sporadic colorectal cancer and adjacent normal intestinal mucosa were examined by phosphite sequencing for the detection of CP G islands in the promoter region of PTTG1 gene. [methods] the relationship between the pathological and clinical features was further studied. [methods] 40 patients with sporadic colorectal cancer were examined for CpG islands in the promoter region of the PTTG1 gene by phosphite sequencing. The methylation rate of the region was 400 BP. Protein imprinting and real-time quantitative PCR(q RT-PCR were used to detect the expression of PTTG1 protein and m RNA in cancer tissues and adjacent tissues, respectively. To analyze the expression of PTTG1 gene and its clinicopathological characteristics. [results] 1. The PTTG1 gene promoter of PTTG1 gene in 40 patients with colorectal cancer and adjacent normal intestinal mucosa was detected by sulfite sequencing. The methylation rate in tumor tissue group was 19.05 卤4.45, and that in paracancerous group was 68.07 卤3.71, the difference was statistically significant. The methylation rate in PTTG1 promoter region of colorectal cancer was lower than that in paracancerous tissue. The methylation rate of cancer tissue was related to the degree of differentiation, depth of invasion, lymph node metastasis, age, The expression of PTTG1 protein was 2.79 卤0.23 and 1.00 卤0.17 in 40 patients with colorectal cancer and relative normal intestinal mucosa, respectively, and the difference was statistically significant (P 0.05). The high expression of PTTG1 protein and the high expression of PTT G1 protein in the adjacent tissues of rectal cancer and the depth of tumor invasion. The degree of differentiation is related to the age of the patient, Sex, tumor size and lymphatic metastasis. Real-time quantitative PCR(q RT-PCR was used to detect the expression of PTTG1 m RNA in cancer tissues and adjacent normal intestinal mucosa of 40 patients with colorectal cancer. The values of 2- 螖螖 CT were 18.38 卤2.14 and 9.37 卤1.49, respectively. The expression of PTTG1 m RNA in colorectal cancer tissues was significantly higher than that in adjacent tissues. [conclusion] 1. The methylation rate of PTT G1 promoter was lower than that of adjacent tissues. 2. PTTG1 gene was highly expressed in sporadic colorectal cancer tissues. It may provide new ideas for the pathogenesis, diagnosis, treatment and prevention of colorectal cancer.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.34

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