RASSF6基因在彌漫大B細胞淋巴瘤中的甲基化狀態(tài)、mRNA表達及其臨床意義的研究
發(fā)布時間：2018-08-03 18:56
【摘要】：目的:非霍奇金淋巴瘤是血液系統(tǒng)惡性腫瘤之一,其中彌漫大B細胞淋巴瘤(Diffuse large B-cell lymphoma,DLBCL)是非霍奇金淋巴瘤中最常見的一種類型,約占成人淋巴瘤的一半,是一組具有高度異質(zhì)性的侵襲性B細胞淋巴瘤,治療方法主要以化療為主。美羅華聯(lián)合化療方案顯著提高了彌漫大B細胞淋巴瘤的療效,R-CHOP方案被認為是標(biāo)準(zhǔn)治療方案。然而,即便是應(yīng)用標(biāo)準(zhǔn)方案,最終仍有將近40%的患者會復(fù)發(fā)進展。因此,彌漫大B細胞淋巴瘤病因?qū)W研究、基因組學(xué)分析為早期診斷、靶向治療及預(yù)后評估提供了新的方向。彌漫大B細胞淋巴瘤的發(fā)生發(fā)展是多基因、多因素共同作用的結(jié)果,因此明確腫瘤相關(guān)基因是早期診斷及后期治療的關(guān)鍵。研究表明,腫瘤的發(fā)生發(fā)展至少包含遺傳和表觀遺傳兩個層面的調(diào)控,包括癌基因激活、抑癌基因失活等,且表觀遺傳似乎起著更加重要的作用。近年來,表觀遺傳現(xiàn)象在多種腫瘤的發(fā)病機制中被明確,包括DNA甲基化、組蛋白修飾、染色質(zhì)重塑等。研究證實RAS相關(guān)區(qū)域家族6基因(RASSF6)是一種抑癌基因,RASSF6的表達可以通過與K-Ras結(jié)合,協(xié)同抑制腫瘤細胞的增殖,且可通過激活JNK信號通路組織細胞周期進展;另外可通過激活caspase通路和其他通路誘導(dǎo)腫瘤細胞凋亡。有研究顯示在兒童白血病、胃癌、乳腺癌等腫瘤中出現(xiàn)RASSF6基因啟動子區(qū)高甲基化現(xiàn)象,甲基化是導(dǎo)致RASSF6基因失活的重要原因之一。而RASSF6基因在彌漫大B細胞淋巴瘤中的相關(guān)研究尚未見報道。本研究將通過應(yīng)用甲基化特異性聚合酶鏈反應(yīng)(Methylation-Specific PCR,MSP)和實時熒光定量逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(real-time Quantitative Reverse Transcriptase-PCR,QRT-PCR)的方法驗證RASSF6在彌漫大B細胞淋巴瘤中啟動子區(qū)甲基化狀態(tài)及表達情況,分析其與樣本資料的關(guān)系,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)。1應(yīng)用MSP方法檢測彌漫大B細胞淋巴瘤患者和健康人外周血RASSF6基因甲基化狀態(tài),并分析其與臨床資料之間的關(guān)系。2應(yīng)用QRT-PCR方法檢測對比彌漫大B細胞淋巴瘤患者和健康人外周血中的RASSF6基因mRNA的相對表達量,并分析其與臨床資料之間的關(guān)系。3應(yīng)用統(tǒng)計軟件SPSS19.0對數(shù)據(jù)進行統(tǒng)計學(xué)分析。結(jié)果:1 RASSF6基因啟動子區(qū)在彌漫大B細胞淋巴瘤患者和健康人外周血中的甲基化率分別為48.6%(17/35)和14.3%(5/35)。彌漫大B細胞淋巴瘤患者RASSF6基因啟動子區(qū)的甲基化率顯著高于健康人,差異有統(tǒng)計學(xué)意義(P0.05)。2分組分析發(fā)現(xiàn),在彌漫大B細胞淋巴瘤患者中,RASSF6甲基化與Ki-67、LDH、臨床分期相關(guān)(P0.05);與患者的性別、年齡、結(jié)外累及、A/B癥狀、生發(fā)中心(Germinal center origin,GCB)或非生發(fā)中心(Nongerminal center origin,Non-GCB)、β2微球蛋白、骨髓累及、IPI均無關(guān)(P0.05)。3彌漫大B細胞淋巴瘤患者RASSF6基因的mRNA表達量為0.0415±0.0287,顯著低于健康人mRNA的表達量0.0634±0.0427,差異有統(tǒng)計學(xué)意義(P0.05)。4分組分析發(fā)現(xiàn),彌漫大B細胞淋巴瘤患者RASSF6基因mRNA的表達量與患者的IPI、Ki-67、結(jié)外累及相關(guān)(P0.05);與患者的性別、年齡、LDH、A/B癥狀、GCB或Non-GCB、β2微球蛋白、骨髓累及、臨床分期均無關(guān)(P0.05)。5彌漫大B細胞淋巴瘤外周血中RASSF6基因啟動子甲基化陽性的mRNA表達量為0.0306±0.0256,甲基化陰性的mRNA相對表達量平均為0.0518±0.0283,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1彌漫大B細胞淋巴瘤患者RASSF6基因啟動子甲基化率顯著高于健康人,這表明RASSF6基因啟動子甲基化可能是彌漫大B細胞淋巴瘤的發(fā)生機制之一。2彌漫大B細胞淋巴瘤患者RASSF6基因mRNA的表達顯著低于健方法:康人,這表明RASSF6基因可能在彌漫大B細胞淋巴瘤中起抑癌基因的作用。3彌漫大B細胞淋巴瘤患者RASSF6基因啟動子區(qū)甲基化率與其mRNA表達呈負相關(guān),提示RASSF6基因啟動子區(qū)甲基化可能是導(dǎo)致彌漫大B細胞淋巴瘤患者mRNA表達減低的原因之一。4彌漫大B細胞淋巴瘤患者RASSF6基因啟動子區(qū)甲基化與患者的Ki-67、LDH、臨床分期相關(guān),這說明RASSF6基因啟動子甲基化狀態(tài)可能與彌漫大B細胞淋巴瘤的侵襲性及預(yù)后相關(guān)。
[Abstract]:Objective: non Hodgkin's lymphoma is one of the malignant tumors of the blood system. The diffuse large B cell lymphoma (Diffuse large B-cell lymphoma, DLBCL) is the most common type of non Hodgkin's lymphoma, which accounts for about half of the adult lymphoma, and is a group of highly heterogeneous invasive B cell lymphoma. The treatment is mainly by chemotherapy. Mainly. The combined chemotherapy regimen has significantly improved the efficacy of diffuse large B cell lymphoma, and the R-CHOP scheme is considered as a standard treatment. However, even with the application standard scheme, nearly 40% of the patients will continue to recur. Therefore, the etiological study of the diffuse large B cell lymphoma, the genomics analysis as an early diagnosis, and the targeted treatment. The development of the treatment and prognosis provides a new direction. The development of the diffuse large B cell lymphoma is the result of multiple genes and multiple factors. Therefore, it is clear that the tumor related genes are the key to early diagnosis and later treatment. The research shows that the development of the tumor includes at least two levels of genetic and epigenetic regulation, including oncogenes. Activation, inactivation of tumor suppressor genes, and epigenetics seem to play a more important role. In recent years, epigenetic phenomena have been identified in the pathogenesis of various tumors, including DNA methylation, histone modification, chromatin remodeling and so on. The study confirmed that the family 6 gene (RASSF6) of the RAS related region is a tumor suppressor gene, and the expression of RASSF6 can be expressed. Combined with K-Ras, the proliferation of tumor cells can be inhibited, and cell cycle progression can be organized by activating the JNK signaling pathway, and apoptosis of tumor cells can be induced by activating the caspase pathway and other pathways. It is one of the important reasons for the inactivation of RASSF6 gene, and the related research of RASSF6 gene in diffuse large B cell lymphoma has not been reported. This study will be based on the application of methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP) and real-time fluorescent quantitative reverse transcriptase polymerase chain reaction (real-time Quantitative Reve). RSE Transcriptase-PCR, QRT-PCR) method verifies the methylation status and expression of RASSF6 in the promoter region of diffuse large B cell lymphoma, and analyzes its relationship with the sample data. It provides a certain theoretical basis for the pathogenesis, clinical diagnosis and treatment of diffuse large B cell lymphoma, and the.1 application MSP method for the detection of diffuse large B cell lymphoma. The methylation status of RASSF6 gene in peripheral blood of patients and healthy people and analysis of the relationship between them and clinical data.2 application QRT-PCR method to detect the relative expression of RASSF6 gene mRNA in diffuse large B cell lymphoma and healthy human peripheral blood, and to analyze the relationship between the clinical data and the.3 Application Statistics Software SPSS19.0 logarithm. Results: the methylation rates of 1 RASSF6 gene promoter region in diffuse large B cell lymphoma and healthy human peripheral blood were 48.6% (17/35) and 14.3% (5/35) respectively. The methylation rate of RASSF6 gene promoter region of diffuse large B cell lymphoma was significantly higher than that of healthy people, and the difference was statistically significant (P0.05).2 grouping. It was found that in patients with diffuse large B cell lymphoma, RASSF6 methylation was associated with Ki-67, LDH, clinical staging (P0.05), and the sex, age, extranodal involvement, A/B symptoms, Germinal center origin, GCB, or non germinal centers (Nongerminal Center), beta 2 microglobulin, and bone marrow involvement in the patients. The mRNA expression of RASSF6 gene in the patients with diffuse large B cell lymphoma was 0.0415 + 0.0287, which was significantly lower than that of healthy mRNA (0.0634 + 0.0427). The difference was statistically significant (P0.05).4 group analysis found that the expression of RASSF6 gene mRNA in patients with diffuse large B cell lymphoma was associated with the patient's IPI, Ki-67, and extranodal involvement (P0.05). The sex, age, LDH, A/B symptoms, GCB or Non-GCB, beta 2 microglobulin, bone marrow involvement, and clinical stages were not related to (P0.05).5 diffuse large B cell lymphoma in the peripheral blood of RASSF6 gene promoter methylation positive mRNA expression of 0.0306 + 0.0256, methylation negative mRNA relative expression of 0.0518 + 0.0283, the difference was statistically significant (P0). .05) conclusion: the methylation rate of RASSF6 gene promoter of RASSF6 gene in patients with diffuse large B cell lymphoma is significantly higher than that of healthy people. This indicates that the methylation of RASSF6 promoter may be one of the mechanisms of diffuse large B cell lymphoma. The expression of RASSF6 gene mRNA in patients with diffuse large B cell lymphoma is significantly lower than that of the healthy method: Kang people, which indicates the RASSF6 base. The methylation rate of the RASSF6 gene promoter region of the diffuse large B cell lymphoma patients with diffuse large B cell lymphoma is negatively correlated with the mRNA expression, suggesting that the methylation of the RASSF6 gene promoter region may be one of the reasons for the decrease of mRNA expression in patients with diffuse large B cell lymphoma, one of the.4 diffuse B cells. The methylation of the promoter region of the RASSF6 gene in the patients with the tumor is associated with the patient's Ki-67, LDH, and clinical staging. This suggests that the methylation status of the RASSF6 gene promoter may be associated with the invasiveness and prognosis of diffuse large B cell lymphoma.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R733.1
本文編號：2162689
[Abstract]:Objective: non Hodgkin's lymphoma is one of the malignant tumors of the blood system. The diffuse large B cell lymphoma (Diffuse large B-cell lymphoma, DLBCL) is the most common type of non Hodgkin's lymphoma, which accounts for about half of the adult lymphoma, and is a group of highly heterogeneous invasive B cell lymphoma. The treatment is mainly by chemotherapy. Mainly. The combined chemotherapy regimen has significantly improved the efficacy of diffuse large B cell lymphoma, and the R-CHOP scheme is considered as a standard treatment. However, even with the application standard scheme, nearly 40% of the patients will continue to recur. Therefore, the etiological study of the diffuse large B cell lymphoma, the genomics analysis as an early diagnosis, and the targeted treatment. The development of the treatment and prognosis provides a new direction. The development of the diffuse large B cell lymphoma is the result of multiple genes and multiple factors. Therefore, it is clear that the tumor related genes are the key to early diagnosis and later treatment. The research shows that the development of the tumor includes at least two levels of genetic and epigenetic regulation, including oncogenes. Activation, inactivation of tumor suppressor genes, and epigenetics seem to play a more important role. In recent years, epigenetic phenomena have been identified in the pathogenesis of various tumors, including DNA methylation, histone modification, chromatin remodeling and so on. The study confirmed that the family 6 gene (RASSF6) of the RAS related region is a tumor suppressor gene, and the expression of RASSF6 can be expressed. Combined with K-Ras, the proliferation of tumor cells can be inhibited, and cell cycle progression can be organized by activating the JNK signaling pathway, and apoptosis of tumor cells can be induced by activating the caspase pathway and other pathways. It is one of the important reasons for the inactivation of RASSF6 gene, and the related research of RASSF6 gene in diffuse large B cell lymphoma has not been reported. This study will be based on the application of methylation specific polymerase chain reaction (Methylation-Specific PCR, MSP) and real-time fluorescent quantitative reverse transcriptase polymerase chain reaction (real-time Quantitative Reve). RSE Transcriptase-PCR, QRT-PCR) method verifies the methylation status and expression of RASSF6 in the promoter region of diffuse large B cell lymphoma, and analyzes its relationship with the sample data. It provides a certain theoretical basis for the pathogenesis, clinical diagnosis and treatment of diffuse large B cell lymphoma, and the.1 application MSP method for the detection of diffuse large B cell lymphoma. The methylation status of RASSF6 gene in peripheral blood of patients and healthy people and analysis of the relationship between them and clinical data.2 application QRT-PCR method to detect the relative expression of RASSF6 gene mRNA in diffuse large B cell lymphoma and healthy human peripheral blood, and to analyze the relationship between the clinical data and the.3 Application Statistics Software SPSS19.0 logarithm. Results: the methylation rates of 1 RASSF6 gene promoter region in diffuse large B cell lymphoma and healthy human peripheral blood were 48.6% (17/35) and 14.3% (5/35) respectively. The methylation rate of RASSF6 gene promoter region of diffuse large B cell lymphoma was significantly higher than that of healthy people, and the difference was statistically significant (P0.05).2 grouping. It was found that in patients with diffuse large B cell lymphoma, RASSF6 methylation was associated with Ki-67, LDH, clinical staging (P0.05), and the sex, age, extranodal involvement, A/B symptoms, Germinal center origin, GCB, or non germinal centers (Nongerminal Center), beta 2 microglobulin, and bone marrow involvement in the patients. The mRNA expression of RASSF6 gene in the patients with diffuse large B cell lymphoma was 0.0415 + 0.0287, which was significantly lower than that of healthy mRNA (0.0634 + 0.0427). The difference was statistically significant (P0.05).4 group analysis found that the expression of RASSF6 gene mRNA in patients with diffuse large B cell lymphoma was associated with the patient's IPI, Ki-67, and extranodal involvement (P0.05). The sex, age, LDH, A/B symptoms, GCB or Non-GCB, beta 2 microglobulin, bone marrow involvement, and clinical stages were not related to (P0.05).5 diffuse large B cell lymphoma in the peripheral blood of RASSF6 gene promoter methylation positive mRNA expression of 0.0306 + 0.0256, methylation negative mRNA relative expression of 0.0518 + 0.0283, the difference was statistically significant (P0). .05) conclusion: the methylation rate of RASSF6 gene promoter of RASSF6 gene in patients with diffuse large B cell lymphoma is significantly higher than that of healthy people. This indicates that the methylation of RASSF6 promoter may be one of the mechanisms of diffuse large B cell lymphoma. The expression of RASSF6 gene mRNA in patients with diffuse large B cell lymphoma is significantly lower than that of the healthy method: Kang people, which indicates the RASSF6 base. The methylation rate of the RASSF6 gene promoter region of the diffuse large B cell lymphoma patients with diffuse large B cell lymphoma is negatively correlated with the mRNA expression, suggesting that the methylation of the RASSF6 gene promoter region may be one of the reasons for the decrease of mRNA expression in patients with diffuse large B cell lymphoma, one of the.4 diffuse B cells. The methylation of the promoter region of the RASSF6 gene in the patients with the tumor is associated with the patient's Ki-67, LDH, and clinical staging. This suggests that the methylation status of the RASSF6 gene promoter may be associated with the invasiveness and prognosis of diffuse large B cell lymphoma.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R733.1
【參考文獻】
相關(guān)期刊論文 前5條
1 許嶸;張文州;陳琳琳;丁小明;謝志新;林陽君;辛軍;;DNA去甲基化藥物作用機制的研究進展[J];現(xiàn)代藥物與臨床;2017年01期
2 陳爾飛;賀紅娟;劉瑞濤;王檬;楊進;;RASSF6在結(jié)直腸癌中的表達研究[J];基因組學(xué)與應(yīng)用生物學(xué);2015年07期
3 李明;王子安;;復(fù)發(fā)/難治性彌漫大B細胞淋巴瘤治療的新進展[J];中華全科醫(yī)學(xué);2015年06期
4 Hui-Lin Ye;Dou-Dou Li;Qing Lin;Yu Zhou;Quan-Bo Zhou;Bing Zeng;Zhi-Qiang Fu;Wen-Chao Gao;Yi-Min Liu;Rui-Wan Chen;Zhi-Hua Li;Ru-Fu Chen;;Low RASSF6 expression in pancreatic ductal adenocarcinoma is associated with poor survival[J];World Journal of Gastroenterology;2015年21期
5 畢婧;高明陽;王衍富;;Ras相關(guān)區(qū)域家族基因啟動子甲基化改變與腫瘤發(fā)生相關(guān)性的研究進展[J];中華臨床醫(yī)師雜志(電子版);2012年16期
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