本文選題：胃癌　切入點：基因甲基　出處：《河北醫(yī)科大學》2016年碩士論文　論文類型：學位論文

【摘要】：胃癌(gastric cancer)占胃部惡性腫瘤的95%以上,其病死率在全球范圍內居惡性腫瘤的第2位,2012年數(shù)據(jù),我國胃癌發(fā)病率占全球43%,死亡率占45%,而早期胃癌僅占10%左右。胃癌在我國仍是最常見的惡性腫瘤之一,是威脅人類健康的一類常見疾病。大量最新研究結果充分表明了基因組學與表觀遺傳學共同機制在促進腫瘤發(fā)生、發(fā)展過程中的重要性并展現(xiàn)出令人樂觀的臨床應用前景。在基因修飾過程中,甲基化將直接影響著一個基因的活性,這是表觀遺傳修飾過程中極其重要的方式。近年來有學者發(fā)現(xiàn)Wnt信號通路的異常激活與腫瘤的發(fā)生、發(fā)展有著密切的關系。尤其是此過程中基于啟動子甲基化致使Wnt信號通路拮抗因子分泌型卷曲相關蛋白(secreted frizzled-related proteins,Sfrp)的沉默或表達活性的嚴重受抑制促進了腫瘤的形成及發(fā)展的過程。目的:本研究旨在探討胃癌組織及對應的癌旁正常組織中Sfrp5基因甲基化修飾及蛋白表達與臨床病理參數(shù)的關系并探討其臨床意義。方法:1選擇石家莊市第一醫(yī)院外科手術切除并病理證實為胃癌的新鮮癌組織及其新鮮癌旁正常組織各90例,胃癌患者中男性73例,女性17例;年齡46-76歲,平均年齡64歲;組織學分型:高、中分化腺癌35例,低分化55例。淋巴結轉移:其中33例患者發(fā)生了鄰近或遠端淋巴結的受累,57例患者未發(fā)生淋巴結轉移。取材部位:每例患者均在胃癌原發(fā)病變區(qū)和切除標本距離切緣大于5厘米處(正常未發(fā)生癌變)分別取材;新鮮組織置于液氮罐中暫存,另外一塊組織經中性福爾馬林固定,經脫水、透明制成蠟塊,切片、常規(guī)HE染色切片經臨床病理科醫(yī)生的觀察保證取材的準確性。所有患者術前均未經化療和放療,均有完整的臨床和病理資料。不符合上述條件的標本被排除于研究之外。2甲基化特異性PCR(methylation specific PCR,MSP)被用于測定各例患者癌組織及正常組織中Sfrp5基因發(fā)生甲基化的狀況,用免疫組織化學(Envision)法檢測Sfrp5蛋白的表達情況,操作步驟嚴格按照試劑盒說明執(zhí)行。3統(tǒng)計學分析。應用SPSS19.0版統(tǒng)計軟件進行數(shù)據(jù)處理及統(tǒng)計學分析,計數(shù)資料以百分比表示,組間比較采用卡方檢驗和Fisher確切概率法分析,以P0.05認為有統(tǒng)計學差異。結果:1 Sfrp5在胃癌和癌旁正常組織中的蛋白表達和甲基化狀態(tài):免疫組化結果:Sfrp5蛋白在胃癌和癌旁正常組織中的陽性表達率分別為20%(18/90)和90%(81/90),不同組別間數(shù)據(jù)具有顯著統(tǒng)計學差異(P0.05)。甲基化特異性PCR結果:Sfrp5基因在癌組織及癌旁組織中的甲基化頻率分別為82.2%(74/90)和18.9%(17/90),差異有統(tǒng)計學意義(P0.05);2 Sfrp5基因甲基化與臨床病理參數(shù)的關系:Sfrp5基因甲基化頻率在無淋巴結轉移組66.7%(22/33)顯著低于有淋巴結轉移組86.0%(49/57),差異有統(tǒng)計學意義(P0.05);高中分化組中Sfrp5基因的甲基化頻率為71.4%(25/35)低分化組的甲基化率81.8%(45/55),但兩組之間的差異不具有統(tǒng)計學意義(P0.05);大于等于50歲年齡組84.5%(71/84)低于小于50歲年齡組基因甲基化頻率100%(6/6),女性組76.4%(13/17)低于男性組甲基化頻率76.7%(56/73),但均無統(tǒng)計學差異(P0.05)。結論:Sfrp5基因高甲基化是其蛋白表達降低的重要原因并且可能與胃癌的發(fā)生、發(fā)展有關。
[Abstract]:Gastric cancer (gastric cancer) accounted for more than 95% of the stomach cancer, the mortality rate of malignant tumor in the world within the scope of the second, 2012 data, the incidence of gastric cancer in China accounted for 43% of global mortality accounted for 45%, while early gastric cancer accounted for only about 10%. Gastric cancer in our country is still one of the most common malignant tumors, is a kind of common disease threatening human health. A large number of the latest research results show that the genomic and epigenetic common mechanism in promoting tumorigenesis, the importance in the process of development and show the clinical application prospects of optimism. In the process of gene modification, methylation will directly affect the activity of a gene. This is an extremely important epigenetic modification in the process. In recent years, some scholars have found that the abnormal activation of Wnt signaling pathway with tumor occurrence, development has a close relationship. Especially a promoter based on this process The base that Wnt signaling pathway antagonist secreted frizzled related protein (secreted frizzled-related, proteins, Sfrp) or silence the expression activity was inhibited seriously promote the formation and development of tumor. Objective: This study aimed to investigate the methylation of Sfrp5 gene in gastric cancer tissues and corresponding adjacent normal tissue modification and protein expression with the clinicopathological parameters and to explore its clinical significance. Methods: 1. Select the first hospital of Shijiazhuang City surgical resection and pathology of gastric cancer and fresh fresh tissue adjacent normal tissues in 90 cases of gastric cancer patients, 73 were male, 17 were female; the age is 46-76 years old, the average age is 64 years old; histological type high, 35 cases of moderately differentiated adenocarcinoma, poorly differentiated 55 cases. Lymph node metastasis: 33 cases occurred in patients with adjacent or distant lymph node involvement, 57 patients without lymph node metastasis. The Ministry of materials A: all patients were in primary gastric lesions and resection margin distance greater than 5 cm (normal non cancerous) were drawn; fresh tissue in liquid nitrogen tank storage, another piece of tissue was fixed with formalin, dehydrated, transparent, paraffin slice, routine HE staining by to observe the clinical pathologist to ensure the accuracy of sampling. All the patients were not treated by chemotherapy and radiotherapy, the clinical and pathological data were complete. Do not meet the above conditions were excluded.2 methylation specific PCR (methylation specific PCR, MSP) was used to determine the methylation of Sfrp5 patients cancer tissue and normal tissue, gene, immunohistochemistry (Envision) method for detection of expression of Sfrp5 protein, operating procedures in strict accordance with the kit instructions.3 application SPSS19.0 version statistics statistical analysis. Software for data processing and statistical analysis, count data expressed as a percentage, comparison between groups using chi square test and Fisher exact probability analysis, to P0.05 that there was statistically significant difference. Results: 1 Sfrp5 in gastric carcinoma and adjacent normal tissues, protein expression and methylation state: the results of immunohistochemistry: the positive expression of Sfrp5 protein in gastric carcinoma and adjacent normal tissues were respectively 20% (18/90) and 90% (81/90), the data between different groups with significant difference (P0.05). Results: the methylation specific PCR methylation frequency of Sfrp5 gene in cancer tissues and in respectively 82.2% and 18.9% (74/90) (17/90), the difference was statistically significant (P0.05); 2 the relationship between Sfrp5 gene methylation and clinicopathological parameters: Sfrp5 gene methylation frequency in the group without lymph node metastasis 66.7% (22/33) with lymph node metastasis was significantly lower than that of group 86% (49/57), the difference was statistically Statistically significant (P0.05); the methylation frequency of Sfrp5 gene differentiated group was 71.4% (25/35) methylation rate of 81.8% low differentiation group (45/55), but the difference between the two groups was not statistically significant (P0.05); greater than or equal to 50 year old age group 84.5% (71/84) lower than that of less than 50 year old age group methylation frequency of 100% (6/6), female group 76.4% (13/17) lower than the male group. The methylation frequency of 76.7% (56/73), but there was no significant difference (P0.05). Conclusion: Sfrp5 gene methylation is an important cause of reduced protein expression and may be related to gastric carcinogenesis and development.

【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R735.2

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