结直肠癌与SLC5A8基因甲基化

发布时间：2018-05-23 08:42

本文选题：结直肠癌 + SLC5A8　；参考：《河北大学》2017年硕士论文

【摘要】：目的:通过探究SLC5A8基因在结直肠癌组织中的表达情况及甲基化现象,为进一步探究SLC5A8基因在结直肠癌发生发展中的作用,为SLC5A8基因甲基化对结直肠癌患者预后评估的相关研究奠定基础。方法:本研究共收集自2015-2016年50例结直肠癌患者标本,其中直肠癌患者为26例,结肠癌患者为24例,男29例,女21例,年龄52-80岁,平均年龄60.5岁,收集一般资料。采集其结直肠癌组织、癌旁组织(距肿瘤边缘0.5cm)、正常组织(距肿瘤边缘5cm以上),贮存在液氮中备用。所有患者术前均未行任何放化疗治疗及免疫治疗。将提取出来的DNA按Epi Tect Fast DNA Kit试剂盒说明书将DNA进行亚硫酸氢盐修饰,用Epi Tect?MSP试剂盒进行甲基化特异性PCR。扩增完成后,取扩增产物进行电泳。将琼脂糖凝胶板放入全自动凝胶成像仪中,在紫外光下显影,拍照保存,并记录实验结果。应用SPSS19.0统计软件对所有数据进行统计学分析处理,定量资料应用方差分析检验,计数资料应用Fisher's确切概率法或卡方检验,检验标准为P0.05代表组间差异有显著统计学意义。结果:1.SLC5A8甲基化基因阳性率在各年龄组间、各性别组间、各发病部位组间以及不同Dukes’临床分期组间未见显著统计学差异(P0.05)。2.50例组织SLC5A8基因甲基化PCR结果显示:癌组织标本有72%(36/50)显示出甲基化扩增产物条带;癌旁组织中为10%(5/50);正常结直肠组织中为2%(1/50)。癌组织的甲基化扩增产物条带明显多于癌旁组织及正常结直肠组织。结直肠癌组织、癌旁组织及正常组织SLC5A8基因甲基化阳性率有明显的差异,且差异有统计学意义(x2=-34.68,P0.05)。进一步两两比较,癌组织SLC5A8基因甲基化阳性率明显高于癌旁组织(72%vs.10%,x2=32.24,P0.05),正常组织(72%vs.2%,x2=45.88,P0.05),但癌旁组织与正常组织之间的SLC5A8基因甲基化阳性率无明显差异(10%vs.2%,P0.05)。36例SLC5A8甲基化的结直肠癌组织中,有2例同时检测到非甲基化的SLC5A8基因,即为部分甲基化;而在SLC5A8基因没有甲基化的结直肠癌组织中,有3例同样没有检测到非甲基化的SLC5A8基因。3.在50例结直肠癌组织中有36例发生了SLC5A8基因甲基化,在这36例阳性标本中有34例(94.44%)未检测到SLC5A8表达。SLC5A8甲基化阴性的14个标本中检测到12例(85.71%)有表达,两者存在显著差异(x2=26.130,P0.05)。SLC5A8的表达与其甲基化状态成负相关(r=-0.682)。结论:1.SLC5A8基因表达的沉默可能参与结直肠癌的发生,在癌组织中的表达明显较癌旁组织及正常组织降低。2.SLC5A8基因的沉默可能与其甲基化相关,基因的甲基化在癌组织中较癌旁组织及正常组织明显增高。3.SLC5A8基因在结直肠癌发生发展中的起到了重要的作用,SLC5A8基因甲基化为结直肠癌患者预后评估的相关研究奠定了基础。
[Abstract]:Objective: to investigate the expression and methylation of SLC5A8 gene in colorectal cancer, and to explore the role of SLC5A8 gene in the carcinogenesis and development of colorectal cancer. To lay a foundation for the study of SLC5A8 gene methylation in prognosis evaluation of colorectal cancer patients. Methods: a total of 50 colorectal cancer specimens were collected from 2015-2016, including 26 cases of rectal cancer, 24 cases of colon cancer, 29 males and 21 females, aged 52-80 years, with an average age of 60.5 years. The tissues of colorectal cancer, paracancerous tissues (0.5 cm from the margin of the tumor) and normal tissues (above 5cm from the margin of the tumor) were collected and stored in liquid nitrogen. All patients were not treated with radiotherapy, chemotherapy or immunotherapy before operation. The extracted DNA was modified with bisulfite according to the specification of Epi Tect Fast DNA Kit kit and methylated by Epi Tect?MSP kit. After the amplification, the amplified products were electrophoretic. The agarose gel plate was put into the automatic gel imager, developed under ultraviolet light, photographed and saved, and the experimental results were recorded. All the data were analyzed and processed by SPSS19.0 statistical software, the quantitative data were analyzed by ANOVA, and the count data were analyzed by Fisher's exact probability method or chi-square test. Results: 1. The positive rate of SLC5A8 methylation gene was found in all age groups and sex groups. There was no significant difference among the groups of different sites and different Dukes' clinical stages. There was no significant difference between the two groups. The results of methylation PCR of SLC5A8 gene in the tissues of 50 cases were as follows: 72 samples of cancer tissues showed the bands of methylation amplification products; It was 10 / 50 / 50 in paracancerous tissues and 2 / 50 / 50 in normal colorectal tissues. The bands of methylation products in cancer tissues were significantly more than those in adjacent tissues and normal colorectal tissues. The positive rate of SLC5A8 gene methylation in colorectal cancer tissues, paracancerous tissues and normal tissues was significantly different, and the difference was statistically significant (P 0.05). Further comparison, the positive rate of SLC5A8 gene methylation in cancer tissues was significantly higher than that in paracancerous tissues (72vs.10x22.24m P0.05), and in normal tissues (72vs.2x2x2c45.88) P0.05. however, there was no significant difference in the methylation positive rate of SLC5A8 gene between adjacent tissues and normal tissues. There was no significant difference in the methylation rate of SLC5A8 gene between the adjacent tissues and the normal tissues. There was no significant difference in the methylation of SLC5A8 gene between the adjacent tissues and the normal tissues in 36 cases of colorectal cancer with SLC5A8 methylation. Unmethylated SLC5A8 gene was detected in 2 cases, that is, partial methylation, while in 3 cases of colorectal cancer without SLC5A8 gene methylation, unmethylated SLC5A8 gene. 3 was also detected. SLC5A8 gene methylation was found in 36 of 50 colorectal cancer tissues and in 34 of the 36 positive specimens. No SLC5A8 expression was detected in 14 specimens with negative methylation of SLC5A8, and in 12 of the 14 samples with negative methylation of SLC5A8, the expression of SLC5A8 was detected in 12 of the 14 specimens with negative methylation of SLC5A8. There was a significant difference between the two groups. The expression of P0.05A8 was negatively correlated with its methylation status. Conclusion the silencing of SLC5A8 gene expression may be involved in the occurrence of colorectal cancer. The expression of SLC5A8 gene in cancer tissues is significantly lower than that in adjacent tissues and normal tissues. 2. The silencing of SLC5A8 gene may be related to its methylation. The methylation of SLC5A8 gene in cancer tissues was significantly higher than that in adjacent tissues and normal tissues. 3. SLC5A8 gene played an important role in the occurrence and development of colorectal cancer.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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