生物钟基因Bmal1、Per2在儿童急性淋巴细胞白血病的表达及意义

发布时间：2018-05-17 23:27

本文选题：急性淋巴细胞白血病 + 儿童　；参考：《郑州大学》2014年硕士论文

【摘要】：研究背景急性淋巴细胞白血病（acute lymphoid leukemia，ALL）是儿童时期最常见的血液肿瘤，占儿童白血病的70%左右，目前发病机制尚不明确。而有研究发现控制生物节律的内在生物钟基因表达异常与肿瘤的发生相关，在急慢性髓系白血病及非霍奇金淋巴瘤中都有检测到钟基因表达异常。现有关钟基因表达与急性淋巴细胞白血病发病关系的研究较少见。研究目的通过对急性淋巴细胞白血病患儿生物钟基因Bmal1、Per2基因表达的检测，探讨生物钟基因在儿童急性淋巴细胞白血病中的表达，初步揭示钟基因在急性淋巴细胞白血病发病中的作用。研究对象和方法 1.选择郑州大学第一附属医院儿童血液科ALL初诊未治疗患儿30例为实验组1（ALL初治组），ALL化疗完全缓解患儿30例为实验组2（ALL-CR组），体检中心体检儿童30例为健康对照组。实验组入选标准参照2006年“儿童急性淋巴细胞白血病诊疗建议（第3次修订草案）”诊断分型标准，实验组和对照组均排除肥胖、心血管疾病、肿瘤及睡眠周期紊乱等影响钟基因表达的疾病。 2.于晨06:00用无菌注射器分别抽取实验组和对照组空腹静脉血各2ml，并迅速打入EDTA抗凝管中，轻轻摇匀。 3.应用密度梯度离心法分离外周血单个核细胞，，吸取白膜层。 4.按照Trizol试剂说明书提取外周血单个核细胞总RNA，检测RNA的纯度及完整性；应用反转录试剂盒合成cDNA；采用Real-time PCR测定Bmal1mRNA、Per2mRNA的表达。结果 1. ALL初治组患儿Bmal1mRNA、Per2mRNA表达与健康对照组比较，差异有统计学意义（P0.05），ALL初治组患儿Per2mRNA表达量水平均较对照组减低，Bmal1mRNA表达水平均较健康对照组增高。 2. ALL-CR组患儿Bmal1mRNA、Per2mRNA表达与健康对照组比较差异有统计学意义（P0.05），ALL-CR组Bmal1mRNA、Per2mRNA表达均较健康对照组减低。 3. ALL-CR组患儿Bmal1mRNA、Per2mRNA表达与ALL初治组比较，差异有统计学意义（P0.05），ALL-CR组Bmal1mRNA、Per2mRNA表达均较ALL初治组减低。结论 1.在儿童ALL外周血单个核细胞中存在生物钟基因Bmal1mRNA、Per2mRNA异常表达,表明其可能在儿童ALL的发生、发展中发挥一定作用； 2.化疗药物应用可影响钟基因的表达。
[Abstract]:Research background Acute lymphoid leukemia (ALL) is the most common blood tumor in childhood, accounting for about 70% of childhood leukemia. Some studies have found that the abnormal expression of the internal clock gene in biological rhythms is related to the occurrence of tumor. The abnormal expression of clock gene has been detected in both acute and chronic myeloid leukemia and non-Hodgkin 's lymphoma. There are few studies on the relationship between bell gene expression and acute lymphoblastic leukemia. Research purpose The expression of biological clock gene (Bmal1PER2) in children with acute lymphoblastic leukemia (AML) was studied, and the role of clock gene in the pathogenesis of acute lymphoblastic leukemia (ALL) was preliminarily revealed. Research objects and methods 1. Thirty untreated children with ALL in Department of Children's Hematology, first affiliated Hospital of Zhengzhou University were selected as experimental group, 30 children with complete remission of all chemotherapy in 1(ALL group, 30 children with complete remission of all chemotherapy in experimental group, and 30 children in physical examination center as healthy control group. According to the criteria for diagnosis and treatment of childhood acute lymphoblastic leukemia (the third revised draft) in 2006, the experimental group and the control group were excluded from obesity and cardiovascular disease. Cancer and sleep cycle disorders affect the expression of clock gene diseases. 2. At 06:00, the fasting venous blood of the experimental group and the control group were extracted with sterile syringe 2 ml each, and the venous blood was quickly injected into the EDTA anticoagulant tube and gently shaken. 3. The peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation. 4. Total RNAs of peripheral blood mononuclear cells were extracted according to Trizol reagent instructions to detect the purity and integrity of RNA. Reverse transcription kit was used to synthesize cDNA. Real-time PCR was used to detect the expression of Bmal1mRNA-Per2 mRNA. Result 1. The expression of Bmal1mRNA-Per2 mRNA in the ALL group was significantly higher than that in the control group (P 0.05). The expression of Bmal1 mRNA in the primary treatment group was significantly higher than that in the control group. 2. The expression of Bmal1mRNA-Per2 mRNA in ALL-CR group was significantly lower than that in healthy control group. The expression of Bmal1mRNA-Per2 mRNA in ALL-CR group was significantly lower than that in healthy control group. 3. The expression of Bmal1mRNA-Per2mRNA in ALL-CR group was significantly lower than that in ALL group. The expression of Bmal1mRNA-Per2mRNA in ALL-CR group was lower than that in ALL group. Conclusion 1. The abnormal expression of Bmal1mRNA-Per2 mRNA in peripheral blood mononuclear cells of children with ALL suggests that Bmal1mRNA-Per2 mRNA may play a role in the occurrence and development of ALL in children. 2. The application of chemotherapeutic drugs can affect the expression of bell gene.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R733.71

【参考文献】