生命早期能量缺乏结直肠癌患者临床病理学特征、临床结局及相关机制探讨
发布时间：2018-08-30 14:50
【摘要】：背景:有研究表明生命早期经历能量缺乏可以降低结直肠癌的发生的几率,其中涉及了表观遗传学的改变,主要是对Cp G岛甲基化表型(Cp Gislandmethylatorphenotype,CIMP)的状态产生影响,而同时有研究显示,肠道微生态的结构变化对患者表观遗传学存在一定的影响,其能通过某些机制使表观遗传学在人体生长发育过程中发生改变。目的:本研究旨在观察生命早期能量缺乏对结直肠癌患者的临床病理学特征及预后的影响,并探讨微生态、微卫星不稳定及表观遗传学等变化在这一过程中所起到的作用。方法:收集1956年~1964年间出生的结直肠癌患者根据其出生时间的不同进行分组,肿瘤标本进行DNA提取,并对DNA进行16Sr DNA的V1-V3高可变区高通量测序,使用主坐标分析法、聚类分析和kruskal-wallis秩和检验分析其肠道微生态分布的多样性及物种丰度的改变。同时,我们随机在三组中抽取标本,合计60个,使用荧光PCR进行BRAF突变的检测,使用毛细管电泳法配合RT-PCR进行微卫星不稳定(MSI)状态的测定,同时,使用甲基化MSP检测(Methylation Specific PCRDetection)进行Cp G岛甲基化表型(Cp Gislandmethylatorphenotype,CIMP)检测。统计学使用t检验以及卡方检验验证患者临床病理学资料以及分子表型与微生态之间的相关性。同时应用双模表示展示在CIMP中MSI和BRAF突变的状态。应用Kaplan-Meier法对三组人群之间的生存率进行分析,了解上述的这一些变化是否会对生存产生不同的影响,以评估能量变化对人体造成的影响。结果:我们发现在不同的组之间在CRC的位置(P=0.034)和CEA水平(P=0.036)以及居住地区的不同(P=0.007)显示出其存在统计学上的差异。同时,三组人群的生存比较发现,相较于非饥荒组,饥荒组的人群5年总生存率高于非饥荒组(OS:68.8%vs 64.2%/62.1%)而在高通量测序显示在所有检测的标本中,我们可以发现具核梭杆菌,脆弱拟杆菌和大肠杆菌在饥荒前组和饥荒后组中丰度较高,而普雷沃氏菌菌在饥荒组中显示更高的丰度,两者的差异情况主要在,其中主要在β多样性中产生。观察到BRAF高突变频率以及高程度的微卫星不稳定较易出现在高程度的CIMP中。脆弱拟杆菌的丰度值与BRAF突变程度存在相关性,大肠杆菌与MSI的突变相关。此外,聚合梭杆菌与CIMP和MSI的状态存在相关性。结论:生命早期能量缺乏的结直肠癌患者呈现生存上的优势。而通过本研究我们发现能量摄入改变可能通过改变肠道微生态某些成分发生变化进而改变CIMP、MSI和BRAF水平来影响CRC患者的预后。
[Abstract]:Background: studies have shown that early life experiences with energy deficiency may reduce the risk of colorectal cancer, involving epigenetic changes, primarily affecting the status of Cp G island methylation phenotype (Cp Gislandmethylatorphenotype,CIMP). At the same time, some studies have shown that the structural changes of intestinal microecology have a certain effect on the epigenetics of patients, which can make epigenetics change in the course of human growth and development through some mechanisms. Objective: to investigate the effect of early life energy deficiency on clinicopathological features and prognosis of colorectal cancer patients, and to explore the role of microecology, microsatellite instability and epigenetic changes in the process. Methods: patients with colorectal cancer born from 1956 to 1964 were divided into groups according to their birth time. DNA was extracted from tumor samples and DNA was sequenced by high throughput of V1-V3 high variable region of 16Sr DNA. Principal coordinate analysis was used. The diversity of intestinal microecological distribution and the change of species abundance were analyzed by cluster analysis and kruskal-wallis rank sum test. At the same time, we randomly sampled 60 samples from three groups. We used fluorescent PCR to detect BRAF mutation, capillary electrophoresis combined with RT-PCR to determine the unstable (MSI) status of microsatellite. Methylation MSP was used to detect (Methylation Specific PCRDetection) for Cp G island methylation phenotype (Cp Gislandmethylatorphenotype,CIMP). T test and chi-square test were used to verify the correlation of clinicopathological data and molecular phenotype with microecology. At the same time, the states of MSI and BRAF mutations in CIMP are demonstrated by using double mode representation. The survival rate of the three groups was analyzed by Kaplan-Meier method to determine whether these changes would have different effects on survival and to evaluate the effects of energy changes on the human body. Results: we found that there were statistical differences between different groups in the location of CRC (P0. 034), CEA level (P0. 036) and living area (P0. 007). At the same time, the survival rate of the three groups was higher than that of the non-famine group (OS:68.8%vs 64.2% / 62.1%). The abundance of Bacteroides fragilis and Escherichia coli was higher in pre-famine group and post-famine group, while that of Prevo-bacillus was higher in famine group. The difference between them was mainly in 尾 -diversity. It was observed that the high mutation frequency of BRAF and the high degree of microsatellite instability were more likely to occur in the high degree of CIMP. The abundance of Bacteroides fragilis was correlated with the mutation of BRAF and the mutation of MSI with Escherichia coli. In addition, there was a correlation between Clostridium Polymerization and the status of CIMP and MSI. Conclusion: colorectal cancer patients with early energy deficiency have the advantage of survival. In this study, we found that the change of energy intake may affect the prognosis of patients with CRC by changing some components of intestinal microecology and then changing the levels of CIMP,MSI and BRAF.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34
本文编号：2213400
[Abstract]:Background: studies have shown that early life experiences with energy deficiency may reduce the risk of colorectal cancer, involving epigenetic changes, primarily affecting the status of Cp G island methylation phenotype (Cp Gislandmethylatorphenotype,CIMP). At the same time, some studies have shown that the structural changes of intestinal microecology have a certain effect on the epigenetics of patients, which can make epigenetics change in the course of human growth and development through some mechanisms. Objective: to investigate the effect of early life energy deficiency on clinicopathological features and prognosis of colorectal cancer patients, and to explore the role of microecology, microsatellite instability and epigenetic changes in the process. Methods: patients with colorectal cancer born from 1956 to 1964 were divided into groups according to their birth time. DNA was extracted from tumor samples and DNA was sequenced by high throughput of V1-V3 high variable region of 16Sr DNA. Principal coordinate analysis was used. The diversity of intestinal microecological distribution and the change of species abundance were analyzed by cluster analysis and kruskal-wallis rank sum test. At the same time, we randomly sampled 60 samples from three groups. We used fluorescent PCR to detect BRAF mutation, capillary electrophoresis combined with RT-PCR to determine the unstable (MSI) status of microsatellite. Methylation MSP was used to detect (Methylation Specific PCRDetection) for Cp G island methylation phenotype (Cp Gislandmethylatorphenotype,CIMP). T test and chi-square test were used to verify the correlation of clinicopathological data and molecular phenotype with microecology. At the same time, the states of MSI and BRAF mutations in CIMP are demonstrated by using double mode representation. The survival rate of the three groups was analyzed by Kaplan-Meier method to determine whether these changes would have different effects on survival and to evaluate the effects of energy changes on the human body. Results: we found that there were statistical differences between different groups in the location of CRC (P0. 034), CEA level (P0. 036) and living area (P0. 007). At the same time, the survival rate of the three groups was higher than that of the non-famine group (OS:68.8%vs 64.2% / 62.1%). The abundance of Bacteroides fragilis and Escherichia coli was higher in pre-famine group and post-famine group, while that of Prevo-bacillus was higher in famine group. The difference between them was mainly in 尾 -diversity. It was observed that the high mutation frequency of BRAF and the high degree of microsatellite instability were more likely to occur in the high degree of CIMP. The abundance of Bacteroides fragilis was correlated with the mutation of BRAF and the mutation of MSI with Escherichia coli. In addition, there was a correlation between Clostridium Polymerization and the status of CIMP and MSI. Conclusion: colorectal cancer patients with early energy deficiency have the advantage of survival. In this study, we found that the change of energy intake may affect the prognosis of patients with CRC by changing some components of intestinal microecology and then changing the levels of CIMP,MSI and BRAF.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34
【参考文献】
相关期刊论文 前1条
1 Harris Bernstein;Carol Bernstein;Claire M Payne;Katerina Dvorak;;Bile acids as endogenous etiologic agents in gastrointestinal cancer[J];World Journal of Gastroenterology;2009年27期
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