生命早期能量缺乏結(jié)直腸癌患者臨床病理學(xué)特征、臨床結(jié)局及相關(guān)機(jī)制探討
發(fā)布時(shí)間：2018-08-30 14:50
【摘要】：背景:有研究表明生命早期經(jīng)歷能量缺乏可以降低結(jié)直腸癌的發(fā)生的幾率,其中涉及了表觀遺傳學(xué)的改變,主要是對(duì)Cp G島甲基化表型(Cp Gislandmethylatorphenotype,CIMP)的狀態(tài)產(chǎn)生影響,而同時(shí)有研究顯示,腸道微生態(tài)的結(jié)構(gòu)變化對(duì)患者表觀遺傳學(xué)存在一定的影響,其能通過某些機(jī)制使表觀遺傳學(xué)在人體生長(zhǎng)發(fā)育過程中發(fā)生改變。目的:本研究旨在觀察生命早期能量缺乏對(duì)結(jié)直腸癌患者的臨床病理學(xué)特征及預(yù)后的影響,并探討微生態(tài)、微衛(wèi)星不穩(wěn)定及表觀遺傳學(xué)等變化在這一過程中所起到的作用。方法:收集1956年~1964年間出生的結(jié)直腸癌患者根據(jù)其出生時(shí)間的不同進(jìn)行分組,腫瘤標(biāo)本進(jìn)行DNA提取,并對(duì)DNA進(jìn)行16Sr DNA的V1-V3高可變區(qū)高通量測(cè)序,使用主坐標(biāo)分析法、聚類分析和kruskal-wallis秩和檢驗(yàn)分析其腸道微生態(tài)分布的多樣性及物種豐度的改變。同時(shí),我們隨機(jī)在三組中抽取標(biāo)本,合計(jì)60個(gè),使用熒光PCR進(jìn)行BRAF突變的檢測(cè),使用毛細(xì)管電泳法配合RT-PCR進(jìn)行微衛(wèi)星不穩(wěn)定(MSI)狀態(tài)的測(cè)定,同時(shí),使用甲基化MSP檢測(cè)(Methylation Specific PCRDetection)進(jìn)行Cp G島甲基化表型(Cp Gislandmethylatorphenotype,CIMP)檢測(cè)。統(tǒng)計(jì)學(xué)使用t檢驗(yàn)以及卡方檢驗(yàn)驗(yàn)證患者臨床病理學(xué)資料以及分子表型與微生態(tài)之間的相關(guān)性。同時(shí)應(yīng)用雙模表示展示在CIMP中MSI和BRAF突變的狀態(tài)。應(yīng)用Kaplan-Meier法對(duì)三組人群之間的生存率進(jìn)行分析,了解上述的這一些變化是否會(huì)對(duì)生存產(chǎn)生不同的影響,以評(píng)估能量變化對(duì)人體造成的影響。結(jié)果:我們發(fā)現(xiàn)在不同的組之間在CRC的位置(P=0.034)和CEA水平(P=0.036)以及居住地區(qū)的不同(P=0.007)顯示出其存在統(tǒng)計(jì)學(xué)上的差異。同時(shí),三組人群的生存比較發(fā)現(xiàn),相較于非饑荒組,饑荒組的人群5年總生存率高于非饑荒組(OS:68.8%vs 64.2%/62.1%)而在高通量測(cè)序顯示在所有檢測(cè)的標(biāo)本中,我們可以發(fā)現(xiàn)具核梭桿菌,脆弱擬桿菌和大腸桿菌在饑荒前組和饑荒后組中豐度較高,而普雷沃氏菌菌在饑荒組中顯示更高的豐度,兩者的差異情況主要在,其中主要在β多樣性中產(chǎn)生。觀察到BRAF高突變頻率以及高程度的微衛(wèi)星不穩(wěn)定較易出現(xiàn)在高程度的CIMP中。脆弱擬桿菌的豐度值與BRAF突變程度存在相關(guān)性,大腸桿菌與MSI的突變相關(guān)。此外,聚合梭桿菌與CIMP和MSI的狀態(tài)存在相關(guān)性。結(jié)論:生命早期能量缺乏的結(jié)直腸癌患者呈現(xiàn)生存上的優(yōu)勢(shì)。而通過本研究我們發(fā)現(xiàn)能量攝入改變可能通過改變腸道微生態(tài)某些成分發(fā)生變化進(jìn)而改變CIMP、MSI和BRAF水平來影響CRC患者的預(yù)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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.34
本文編號(hào)：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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.34
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
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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