本文選題：淋巴瘤 + 表觀遺傳學�。� 參考：《河北醫(yī)科大學》2016年碩士論文

【摘要】：目的:淋巴瘤是最早發(fā)現(xiàn)的血液系統(tǒng)惡性腫瘤之一,近年來其發(fā)病率呈上升趨勢,雖然淋巴瘤的治療方法不斷改善,經(jīng)過聯(lián)合化療后得以延長半數(shù)以上患者的生存期,但仍然有部分患者死于腫瘤的進展。因此關注淋巴瘤病因?qū)W研究、早期診斷及靶向治療是十分必要的。近年來,越來越多的研究表明表觀遺傳在腫瘤的發(fā)生發(fā)展過程中起到重要作用,而DNA甲基化水平改變是目前研究最為廣泛的表觀遺傳學機制之一。RASSF2是RASSF家族中的一員,是一種抑癌基因。RASSF2包括RASSF2A、RASSF2B和RASSF2C三個轉(zhuǎn)錄本。其中RASSF2A具有促進細胞凋亡和抑制生長作用,同時它還能抑制炎癥因子表達,達到抑制腫瘤浸潤的目的。有研究發(fā)現(xiàn),在結腸癌、結腸癌細胞系以及結腸腺瘤中RASSF2A基因啟動子區(qū)普遍存在高甲基化,而在胃癌、子宮內(nèi)膜癌、鼻咽癌、肺癌中也能檢測到RASSF2A啟動子區(qū)甲基化現(xiàn)象的發(fā)生。甲基化是導致RASSF2A基因失活的重要機制。但至今,RASSF2A在彌漫大B細胞淋巴瘤中的相關研究尚未見報道。本研究采用逆轉(zhuǎn)錄-聚合酶鏈反應(Reverse Transcriptase-PCR,RT-PCR)和甲基化特異性聚合酶鏈反應(Methylation-Specific PCR,MSP)的方法,以正常人外周血為對照,檢測彌漫大B細胞淋巴瘤患者外周血中的RASSF2A基因甲基化狀態(tài)及m RNA表達情況,并結合相應的臨床資料分別作相關性分析,為彌漫大B細胞淋巴瘤的發(fā)病機制、臨床診斷及治療提供一定的理論依據(jù)。方法:1應用MSP檢測彌漫大B細胞淋巴瘤患者外周血以及正常人外周血的RASSF2A基因甲基化狀態(tài),并分析該基因甲基化與臨床資料之間的關系。2應用RT-PCR檢測彌漫大B細胞淋巴瘤患者外周血以及正常人外周血的RASSF2A基因m RNA的相對表達量,并分析該基因m RNA相對表達量與臨床資料之間的關系。3采用SPSS 19.0軟件對數(shù)據(jù)進行統(tǒng)計學分析。結果:1 RASSF2A基因在彌漫大B細胞淋巴瘤患者外周血中甲基化率為42.5%(17/40),顯著高于正常人外周血的甲基化率10.0%(4/40),差異有統(tǒng)計學意義(P0.05)。RASSF2A基因在彌漫大B細胞淋巴瘤患者的外周血中的甲基化與患者的Ki-67、結外累及與否以及LDH相關,與正常人外周血相比,差異有統(tǒng)計學意義(P0.05);與患者的IPI評分、骨髓累及與否、年齡、性別、A/B癥狀、生發(fā)中心或非生發(fā)中心、β2微球蛋白以及臨床分期均無關,與正常人外周血相比,差異無統(tǒng)計學意義(P0.05)。2 RASSF2A基因在彌漫大B細胞淋巴瘤患者外周血中的m RNA表達量為0.49±0.13,明顯低于在正常人外周血中的m RNA的表達量0.56±0.17,且差異有統(tǒng)計學意義(P0.05)。RASSF2A基因在彌漫大B細胞淋巴瘤患者外周血中m RNA的表達量與患者IPI評分、結外累及與否、Ki-67以及LDH相關的,與正常人對照組相比,差異有統(tǒng)計學意義(P0.05);與患者的A/B癥狀、年齡、性別、β2微球蛋白、生發(fā)中心或非生發(fā)中心、臨床分期、骨髓累及與否均無關,差異無統(tǒng)計學意義(P0.05)。3彌漫大B細胞淋巴瘤患者的外周血中的RASSF2A基因啟動子的檢測結果表明其甲基化為陽性,其m RNA平均相對表達量為0.41±0.12,甲基化陰性的m RNA平均相對表達量為0.55±0.11,兩者差異有統(tǒng)計學意義(P0.05)。結論:1彌漫大B細胞淋巴瘤患者外周血中RASSF2A的甲基化率明顯高于正常對照組的甲基化率,提示RASSF2A基因啟動子甲基化可能與彌漫大B細胞淋巴瘤發(fā)生相關。2彌漫大B細胞淋巴瘤患者外周血中RASSF2A基因m RNA的表達明顯低于正常人對照組,提示RASSF2A基因可能在彌漫大B細胞淋巴瘤具有抑癌基因的作用。3彌漫大B細胞淋巴瘤患者外周血中RASSF2A基因啟動子呈高甲基化,其m RNA表達降低,提示彌漫大B細胞淋巴瘤中RASSF2A基因甲基化可能與其m RNA表達降低有關。4彌漫大B細胞淋巴瘤患者外周血中,RASSF2A基因啟動子甲基化與患者的結外累及與否、Ki-67和LDH相關,提示RASSF2A基因啟動子甲基化可作為評價彌漫大B細胞淋巴瘤的惡性進程、侵襲性以及預后的指標。
[Abstract]:Objective: lymphoma is one of the earliest found malignant tumors of the blood system. In recent years, the incidence of lymphoma is on the rise. Although the treatment method of lymphoma is continuously improved, the survival period of more than half of the patients can be prolonged after combined chemotherapy, but some patients still die of the progress of the tumor. Therefore, attention is paid to the etiology of lymphoma. Diagnosis and targeted therapy are very necessary. In recent years, more and more studies have shown that epigenetic inheritance plays an important role in the development of tumor. The change of DNA methylation level is one of the most widely studied epigenetic mechanisms,.RASSF2 is a member of the RASSF family, and a tumor suppressor gene,.RASSF2, including RASSF2 Three transcripts of A, RASSF2B and RASSF2C, in which RASSF2A can promote cell apoptosis and inhibit growth, and also inhibit the expression of inflammatory factors to inhibit tumor infiltration. The methylation of RASSF2A promoter region can also be detected in endometrial carcinoma, nasopharyngeal carcinoma, and lung cancer. Methylation is an important mechanism leading to the inactivation of RASSF2A gene. But up to now, the related research of RASSF2A in diffuse large B cell lymphoma has not been reported. This study uses reverse transcriptase polymerase chain reaction (Reverse Transcriptase-PCR, RT-). PCR) and methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP), using normal human peripheral blood as the control, to detect the methylation status of RASSF2A gene and the expression of M RNA in peripheral blood of patients with diffuse large B cell lymphoma, and to combine the corresponding clinical data for the diffuse large B cell lymphoma. The pathogenesis, clinical diagnosis and treatment provided a certain theoretical basis. Methods: 1 MSP was used to detect the methylation of RASSF2A gene in peripheral blood of patients with diffuse large B cell lymphoma and normal human peripheral blood, and the relationship between methylation and clinical data was analyzed by.2 application RT-PCR to detect the peripheral blood of patients with diffuse large B cell lymphoma The relative expression of RASSF2A gene m RNA in blood and normal human peripheral blood, and analysis of the relationship between the relative expression of M RNA and the clinical data.3 using SPSS 19 software to analyze the data statistically. Results: the 1 RASSF2A gene was 42.5% (17/40) in the peripheral blood of the diffuse large B cell lymphoma patients, which was significantly higher than that of 42.5% (17/40). The methylation rate of peripheral blood of normal people was 10% (4/40). The difference was statistically significant (P0.05) the methylation of.RASSF2A gene in peripheral blood of patients with diffuse large B cell lymphoma was associated with the patient's Ki-67, extranodal involvement and LDH, and the difference was statistically significant (P0.05) compared with the normal peripheral blood (P0.05); and the IPI score of the patients, and bone marrow involvement. Whether age, sex, A/B symptoms, germinal center or non germinal center, beta 2 microglobulin and clinical staging were not related, compared with normal human peripheral blood, the difference was not statistically significant (P0.05), the amount of M RNA in peripheral blood of.2 RASSF2A gene in diffuse large B cell lymphoma was 0.49 + 0.13, significantly lower than that of M R in normal peripheral blood. The expression of NA was 0.56 + 0.17, and the difference was statistically significant (P0.05) in the peripheral blood of patients with diffuse large B cell lymphoma, the expression of M RNA was related to the IPI score, Ki-67 and LDH, and the difference was statistically significant compared with the normal control group (P0.05); A/B symptoms, age, sex, and beta 2 were compared with those of the control group. Microglobulin, germinal center or non germinal center, clinical staging and bone marrow involvement were not related, and the difference was not statistically significant (P0.05) the detection of RASSF2A gene promoter in peripheral blood of patients with.3 diffuse large B cell lymphoma showed that the methylation was positive, the average relative expression of M RNA was 0.41 + 0.12, and the methylation negative m RNA The mean relative expression was 0.55 + 0.11, and the difference was statistically significant (P0.05). Conclusion: the methylation rate of RASSF2A in peripheral blood of patients with 1 diffuse large B cell lymphoma was significantly higher than that of normal control group, suggesting that the methylation of RASSF2A gene promoter may be associated with the diffuse large B cell lymphoma of.2 diffuse large B cell lymphoma. The expression of RASSF2A gene m RNA in peripheral blood of patients was significantly lower than that in normal controls, suggesting that the RASSF2A gene might be in the diffuse large B cell lymphoma with tumor suppressor gene, the promoter of RASSF2A gene promoter in the peripheral blood of.3 diffuse large B cell lymphoma was hypermethylation, and the m RNA expression decreased, suggesting that the diffuse large B cell lymphoma was RASSF. The methylation of 2A gene may be associated with the decrease in the expression of M RNA expression in peripheral blood of patients with.4 diffuse large B cell lymphoma. RASSF2A gene promoter methylation is associated with Ki-67 and LDH, which suggests that the RASSF2A gene promoter methylation may be used to evaluate the malignant progression, invasiveness, and prognosis of diffuse large B cells. Mark.
【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】：R733.1

【參考文獻】

相關期刊論文 前3條

1 Hampig Raphael Kourie;Joseph Gharios;Fadi Elkarak;Joelle Antoun;Marwan Ghosn;;Is metastatic pancreatic cancer an untargetable malignancy?[J];World Journal of Gastrointestinal Oncology;2016年03期

2 韓競男;魯昊騁;梁靜;;DNA甲基化與癌癥[J];中國生物化學與分子生物學報;2012年02期

3 江培洲,沈新明,黃華;癌基因與抑癌基因的表達研究進展[J];國外醫(yī)學(腫瘤學分冊);2001年02期

，

本文編號：2067418