摘要
目的:探讨RAS相关区域家族5A(Ras-association domain family 5A,RASSF5A)基因在淋巴瘤患者及正常人外周血中DNA甲基化和mRNA表达状态,并研究其与患者临床表现的关系。方法:应用甲基化特异性PCR(methylation-specific PCR,MSP)及RT-PCR方法检测河北医科大学第四附属医院2013年10月至2015年3月收治的弥漫性大B细胞淋巴瘤患者74例、T细胞淋巴瘤患者42例及健康志愿者42人的外周血中RASSF5A基因甲基化及mRNA表达状态。分析其与临床表现之间的关系。结果:RASSF5A在弥漫性大B细胞淋巴瘤和T细胞淋巴瘤中的甲基化率分别为64.9%(48/74)和73.8%(31/42),明显高于正常人的7.1%(3/42)(P<0.05);而其mRNA表达量分别为0.54±0.17和0.52±0.18,均低于正常人的0.86±0.10(P<0.05)。弥漫性大B细胞淋巴瘤中RASSF5A基因启动子甲基化阳性的mRNA相对表达量(0.51±0.18)低于甲基化阴性的mRNA表达量(0.60±0.17)(P<0.05)。T细胞淋巴瘤中RASSF5A基因启动子甲基化阳性的mRNA相对表达量(0.50±0.15)低于甲基化阴性的mRNA表达量(0.63±0.12)(P<0.05)。RASSF5A基因在弥漫性大B细胞淋巴瘤中的甲基化率与患者的LDH、IPI评分及Ki67相关(P<0.05);RASSF5A基因在T细胞淋巴瘤中的甲基化率与患者的临床分期、结外累及及Ki-67相关(P<0.05)。RASSF5A基因在弥漫性大B细胞淋巴瘤中mRNA的表达量与患者的IPI评分及结外累及相关(P<0.05);RASSF5A基因在T细胞淋巴瘤中mRNA的表达量与患者的临床分期和结外累及相关(P<0.05)。结论:RASSF5A基因启动子甲基化可能是弥漫性大B细胞淋巴瘤和T细胞淋巴瘤发生的机制之一,mRNA表达沉默可能是表观遗传学机制之一。RASSF5A基因在弥漫性大B细胞淋巴瘤和T细胞淋巴瘤中主要可能起抑癌基因的作用,与淋巴瘤的侵袭性、恶性进程及预后有关。
Objective： To investigate the promoter methylation status of Ras-association domain family 5A（ RASSF5A）gene and its mRNA expression level in in the peripheral blood of patients with lymphoma and normal people as well as its correlation to clinical manifestations. Methods： Methylation-specific PCR（ MSP） and Reverse Transcriptase-PCR（ RTPCR） method were used to examine the methylation status and the mRNA expression of RASSF5 A gene in peripheral blood of patients with diffused large B cell lymphoma or T cell lymphoma and healthy participants. The relationship between RASSF5 A gene methylation and clinical data was further analyzed. Results： The methylation frequencies of RASSF5 A in diffused large B cell lymphoma and T cell lymphoma（ 64. 9% [48 /74] and 73. 8% [31 /42]） were significantly higher than that of healthy participants（ 7. 1% [3 /42 ],all P〈0. 05）. The mRNA expressions of RASSF5 A gene in diffused large B cell lymphoma and peripheral T cell lymphoma were（ 0. 54 ± 0. 17） and（ 0. 52 ± 0. 18）,respectively,which were significantly lower than that of healthy participants（ 0. 86 ± 0. 10）（ P〈0. 05）. The mRNA expression of RASSF5 A gene with positive promoter methylation（ [0. 51 ± 0. 18]; [0. 50 ± 0. 15]） was significantly lower than that of the gene with negative methylation（ [0. 60 ± 0. 17]; [0. 63 ± 0. 12]）（ all P〈0. 05） in both diffused large B cell lymphoma and T cell lymphoma. Methylation frequency of RASSF5 A gene was associated with LDH,IPI,and Ki-67（ P〈0. 05） in diffused large B cell lymphoma,and associated with clinical stage,extranodal involvement and Ki-67（ P〈0. 05） in T cell lymphoma. The mRNA expression of RASSF5 A gene was correlated with IPI score,extranodal involvement（ P〈0. 05） in diffused large B cell lymphoma,and correlated with clinical stage and extranodal involvement（ P〈0. 05） in T cell lymphoma. Conclusion： The methylation of RASSF5 A gene may be associated with the occurrence of diffused large B c
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2016年第4期537-544,共8页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81472335)~~
