本文關(guān)鍵詞： 胃癌 MGMT 甲基化 DNMT1　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:本實(shí)驗(yàn)檢測胃癌組織以及癌旁組織中6-氧-甲基鳥嘌呤-DNA甲基轉(zhuǎn)移酶(MGMT)基因的甲基化以及MGMT蛋白的表達(dá)情況,分析MGMT基因甲基化的狀態(tài)與胃癌發(fā)生以及發(fā)展的關(guān)系。探究胃癌組織以及癌旁組織中DNA甲基轉(zhuǎn)移酶1(DNA methyltransferase 1,DNMT1)蛋白表達(dá)情況,分析其與胃癌的關(guān)系及臨床意義。另外還對胃癌組織中MGMT甲基化與DNMT1蛋白表達(dá)的關(guān)系進(jìn)行初步的探究。方法:1.收集青島市市立醫(yī)院普外科于2015年1月至2016年7月切除的60例胃癌患者的新鮮胃癌組織及其相應(yīng)的癌旁組織標(biāo)本,胃癌組織取自癌灶中央非壞死區(qū)域,癌旁組織取自距離癌組織5cm以上,且術(shù)前未行放療、化療、靶向治療等抗腫瘤治療。提取組織DNA,采用甲基化特異性PCR(MSP)技術(shù)檢測上述組織標(biāo)本中MGMT基因甲基化狀態(tài)。分析MGMT基因甲基化與胃癌的關(guān)系。2.提取胃癌組織及相應(yīng)癌旁組織中的RNA,應(yīng)用逆轉(zhuǎn)錄-多聚合酶鏈反應(yīng)(RT-PCR)檢測MGMT及DNMT1的mRNA的表達(dá)情況。3.用免疫組化SP法(IHC)檢測胃癌組織以及相應(yīng)的癌旁組織中MGMT和DNMT1的蛋白表達(dá)情況,綜合分析MGMT及DNMT1表達(dá)與胃癌的關(guān)系,并對胃癌中MGMT基因甲基化及DNMT1蛋白表達(dá)的關(guān)系進(jìn)行初步探討。結(jié)果:1.胃癌組織中MGMT基因甲基化率為45.00%(27/60),明顯高于癌旁組織(13.33%,8/60),差異有意義(χ2=14.56,P0.001)。2.胃癌組織中MGMT mRNA陽性表達(dá)率低于癌旁組織(41.67%vs93.33%,χ2=36.51,P0.001);而胃癌組織中DNMT1 mRNA陽性率明顯高于癌旁組織(76.67%vs18.33%,χ2=40.94,P0.001)。3.MGMT mRNA陰性表達(dá)的胃癌組織中其基因的甲基化率較陽性表達(dá)者升高(57.14%vs28.00%,χ2=11.96,P0.05)。4.胃癌組織以及癌旁組織中MGMT以及DNMT1蛋白表達(dá)情況與mRNA表達(dá)基本一致,MGMT蛋白在胃癌組織中的表達(dá)陽性率(40%,24/60)明顯低于癌旁組織(93.33%,56/60),差異有統(tǒng)計(jì)學(xué)意義(χ2=38.4,P0.001)。而DNMT1在胃癌組織中的表達(dá)陽性率(76.67%,46/60)卻明顯高于癌旁組織(6.67%,4/60),差異有意義(χ2=60.48,P0.001)。5.胃癌組織中MGMT與DNMT1蛋白表達(dá)呈負(fù)相關(guān)(r=-0.795,P0.01)。結(jié)論:1.MGMT基因高甲基化與胃癌的發(fā)生以及發(fā)展有關(guān)。2.MGMT的低表達(dá)以及DNMT1的高表達(dá)在胃癌的發(fā)生過程中起重要作用。3.在胃癌中MGMT高甲基化改變可能是導(dǎo)致MGMT低表達(dá)的原因之一。4.DNMT1可能調(diào)控MGMT基因的甲基化過程。意義:胃癌是一種死亡率很高的惡性腫瘤,大部分胃癌患者確診時(shí)已處于晚期階段,其早期診斷十分重要。胃癌產(chǎn)生與演變過程極為復(fù)雜,涉及環(huán)境、遺傳等各個(gè)方面,其中DNA甲基化在胃癌發(fā)生過程中起到一定作用。本實(shí)驗(yàn)在表觀遺傳學(xué)水平,研究MGMT基因高甲基化及DNMT1蛋白表達(dá)與胃癌的關(guān)系,為胃癌的診斷、腫瘤分期以及預(yù)后等方面提供思路。
[Abstract]:Objective: to detect the methylation of 6-oxy-methylguanine-DNA methyltransferase (MGMT) gene and the expression of MGMT protein in gastric cancer tissues and adjacent tissues. To analyze the relationship between the status of methylation of MGMT gene and the occurrence and development of gastric cancer, and to investigate the expression of DNA methyltransferase 1 methyltransferase 1 methyltransferase 1 DNMT1 protein in gastric cancer tissues and adjacent tissues. In addition, the relationship between MGMT methylation and DNMT1 protein expression in gastric cancer tissues was studied. Methods: 1. Collecting general surgery department of Qingdao Municipal Hospital from January 2015 to July 2016. Fresh gastric cancer tissues and their corresponding paracancerous tissues from 60 patients with gastric cancer resected, The tissues of gastric cancer were taken from the non-necrotic area in the center of the cancer focus, and the adjacent tissues were taken from the tissues more than 5 cm from the cancerous tissue, and they were not treated with radiotherapy and chemotherapy before operation. The methylation status of MGMT gene was detected by methylation specific PCR technique. The relationship between methylation of MGMT gene and gastric cancer was analyzed. 2. The tissues of gastric cancer and corresponding cancer were extracted. The expression of mRNA in MGMT and DNMT1 was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of MGMT and DNMT1 in gastric carcinoma and adjacent tissues was detected by immunohistochemical SP method. The relationship between the expression of MGMT and DNMT1 and gastric cancer was analyzed. The relationship between the methylation of MGMT gene and the expression of DNMT1 protein in gastric cancer was studied. Results: 1. The methylation rate of MGMT gene in gastric cancer was 45.000.27 / 60%, which was significantly higher than that in paracancerous tissue (13.33 / 60 / 60). The difference was significant (蠂 ~ 2 ~ (14. 5) P ~ (0.001) P ~ (0.001) 路2.The positive expression of MGMT mRNA in gastric cancer tissue was higher than that in paracancerous tissue (P ~ (0.001 / 60)). The positive rate of DNMT1 mRNA in gastric cancer tissues was significantly higher than that in paracancerous tissues (76.67 vs 18.33). The methylation rate of genes in gastric cancer tissues with negative expression of mRNA was higher than that in those with positive expression of 57.14 vs 28.00, 蠂 211.96U P 0.05p 0.05. 4. The methylation rate of MGMT in gastric cancer tissues and adjacent tissues was significantly higher than that in cancer tissues with negative expression of VS93.331.The positive rate of methylation in gastric cancer tissues was significantly higher than that in those with negative expression of VS93.330.The positive rate of DNMT1 mRNA in gastric cancer tissues and adjacent tissues was significantly higher than that in gastric cancer tissues and adjacent tissues. The methylation rate of MGMT in gastric cancer tissues and adjacent tissues was significantly higher than that in those with positive expression. The positive rate of DNMT1 protein expression in gastric cancer tissues was significantly lower than that in adjacent tissues 93.33 / 56 / 60 (蠂 ~ 2 / 38.4N / P 0.001), while the positive rate of DNMT1 expression in gastric cancer tissues was 76.6746 / 60), which was significantly higher than that in cancer tissues (76.67 / 46 / 60), and the positive rate of DNMT1 expression in gastric cancer tissues was significantly higher than that in paracancerous tissues (蠂 ~ 2 / 38.4N ~ (56 / 60)) (蠂 ~ (2 / 2) ~ (38.4) P 0.001). The difference was significant (蠂 ~ 2 / 60.48 / P 0.001 ~ 1.5.The expression of MGMT and DNMT1 protein was negatively correlated with the expression of DNMT1 protein. Conclusion: 1. The hypermethylation of MGMT gene is related to the occurrence and development of gastric cancer. 2. The low expression of MGMT and the high expression of DNMT1 may play an important role in the pathogenesis of gastric cancer. The change of MGMT hypermethylation in gastric cancer may be one of the causes of low expression of MGMT. 4. DNMT1 may regulate the methylation process of MGMT gene. Most patients with gastric cancer are in the late stage of diagnosis, and their early diagnosis is very important. The process of gastric cancer production and evolution is extremely complex, involving various aspects, such as environment, heredity and so on. DNA methylation plays an important role in the carcinogenesis of gastric cancer. In this study, the relationship between the hypermethylation of MGMT gene and the expression of DNMT1 protein in gastric cancer was studied to be the diagnosis of gastric cancer. Tumor staging and prognosis provide ideas.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.2

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