本文選題：結(jié)直腸癌 + SLC5A8��； 參考：《河北大學(xué)》2017年碩士論文

【摘要】：目的:通過(guò)探究SLC5A8基因在結(jié)直腸癌組織中的表達(dá)情況及甲基化現(xiàn)象,為進(jìn)一步探究SLC5A8基因在結(jié)直腸癌發(fā)生發(fā)展中的作用,為SLC5A8基因甲基化對(duì)結(jié)直腸癌患者預(yù)后評(píng)估的相關(guān)研究奠定基礎(chǔ)。方法:本研究共收集自2015-2016年50例結(jié)直腸癌患者標(biāo)本,其中直腸癌患者為26例,結(jié)腸癌患者為24例,男29例,女21例,年齡52-80歲,平均年齡60.5歲,收集一般資料。采集其結(jié)直腸癌組織、癌旁組織(距腫瘤邊緣0.5cm)、正常組織(距腫瘤邊緣5cm以上),貯存在液氮中備用。所有患者術(shù)前均未行任何放化療治療及免疫治療。將提取出來(lái)的DNA按Epi Tect Fast DNA Kit試劑盒說(shuō)明書(shū)將DNA進(jìn)行亞硫酸氫鹽修飾,用Epi Tect?MSP試劑盒進(jìn)行甲基化特異性PCR。擴(kuò)增完成后,取擴(kuò)增產(chǎn)物進(jìn)行電泳。將瓊脂糖凝膠板放入全自動(dòng)凝膠成像儀中,在紫外光下顯影,拍照保存,并記錄實(shí)驗(yàn)結(jié)果。應(yīng)用SPSS19.0統(tǒng)計(jì)軟件對(duì)所有數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析處理,定量資料應(yīng)用方差分析檢驗(yàn),計(jì)數(shù)資料應(yīng)用Fisher's確切概率法或卡方檢驗(yàn),檢驗(yàn)標(biāo)準(zhǔn)為P0.05代表組間差異有顯著統(tǒng)計(jì)學(xué)意義。結(jié)果:1.SLC5A8甲基化基因陽(yáng)性率在各年齡組間、各性別組間、各發(fā)病部位組間以及不同Dukes’臨床分期組間未見(jiàn)顯著統(tǒng)計(jì)學(xué)差異(P0.05)。2.50例組織SLC5A8基因甲基化PCR結(jié)果顯示:癌組織標(biāo)本有72%(36/50)顯示出甲基化擴(kuò)增產(chǎn)物條帶;癌旁組織中為10%(5/50);正常結(jié)直腸組織中為2%(1/50)。癌組織的甲基化擴(kuò)增產(chǎn)物條帶明顯多于癌旁組織及正常結(jié)直腸組織。結(jié)直腸癌組織、癌旁組織及正常組織SLC5A8基因甲基化陽(yáng)性率有明顯的差異,且差異有統(tǒng)計(jì)學(xué)意義(x2=-34.68,P0.05)。進(jìn)一步兩兩比較,癌組織SLC5A8基因甲基化陽(yáng)性率明顯高于癌旁組織(72%vs.10%,x2=32.24,P0.05),正常組織(72%vs.2%,x2=45.88,P0.05),但癌旁組織與正常組織之間的SLC5A8基因甲基化陽(yáng)性率無(wú)明顯差異(10%vs.2%,P0.05)。36例SLC5A8甲基化的結(jié)直腸癌組織中,有2例同時(shí)檢測(cè)到非甲基化的SLC5A8基因,即為部分甲基化;而在SLC5A8基因沒(méi)有甲基化的結(jié)直腸癌組織中,有3例同樣沒(méi)有檢測(cè)到非甲基化的SLC5A8基因。3.在50例結(jié)直腸癌組織中有36例發(fā)生了SLC5A8基因甲基化,在這36例陽(yáng)性標(biāo)本中有34例(94.44%)未檢測(cè)到SLC5A8表達(dá)。SLC5A8甲基化陰性的14個(gè)標(biāo)本中檢測(cè)到12例(85.71%)有表達(dá),兩者存在顯著差異(x2=26.130,P0.05)。SLC5A8的表達(dá)與其甲基化狀態(tài)成負(fù)相關(guān)(r=-0.682)。結(jié)論:1.SLC5A8基因表達(dá)的沉默可能參與結(jié)直腸癌的發(fā)生,在癌組織中的表達(dá)明顯較癌旁組織及正常組織降低。2.SLC5A8基因的沉默可能與其甲基化相關(guān),基因的甲基化在癌組織中較癌旁組織及正常組織明顯增高。3.SLC5A8基因在結(jié)直腸癌發(fā)生發(fā)展中的起到了重要的作用,SLC5A8基因甲基化為結(jié)直腸癌患者預(yù)后評(píng)估的相關(guān)研究奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.34

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