Atg5和Atg7在儿童急性白血病中的表达及相关分析

发布时间：2018-01-19 08:48

本文关键词： 急性白血病自噬免疫组织化学染色 Atg5 Atg7 蛋白　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:急性白血病(acute leukemia,AL)是一类常见的造血干细胞恶性增殖性疾病,目前认为自噬调控失调与肿瘤发生密切相关。自噬是真核细胞在自噬相关基因(autophagy related gene,Atg)的调控下利用溶酶体降解受损细胞器和大分子物质的过程,在维持细胞稳定方面起重要作用,是细胞适应内外环境所需的一种基本调节方式。采用免疫组织化学染色法对儿童急性白血病骨髓细胞中自噬相关基因Atg5、Atg7蛋白表达水平进行检测,探讨这两种因子活性与儿童急性白血病发生、发展的关系。方法1研究对象:实验组随机选取自2014年1月至2016年6月河北医科大学第二医院儿科住院的初诊未治急性白血病患儿50例,其中急性淋巴细胞白血病组30例,急性髓细胞白血病组20例,对照组选取同期与实验组年龄性别相匹配的非恶性血液病(包括免疫性血小板减少症、营养性贫血)患儿20例。2标本采集和处理:分别抽取实验组及对照组新鲜骨髓标本2ml,自然凝固后经4%多聚甲醛固定,常规石蜡包埋、切片。3实验方法:采用免疫组织化学染色法检测儿童急性淋巴细胞白血病、急性髓细胞白血病及对照组骨髓细胞中Atg5、Atg7蛋白的表达水平。4统计学分析:应用SPSS13.0统计分析软件进行统计学分析,Atg5、Atg7蛋白在各组标本中表达强度采用x2检验或Fisher确切概率法,P0.05有统计学意义;Atg5、Atg7蛋白的相关性使用Spearman相关性分析进行检验,P0.01有统计学意义。结果1 Atg5蛋白的表达情况1.1 Atg5在急性白血病中阳性率(82.0%)高于对照组阳性率(40.0%),两者之间表达差异有统计学意义(x2=12.0,P0.05)。1.2 Atg5在急性淋巴细胞白血病中阳性率为83.3%,在急性髓细胞白血病中阳性率80.0%,分别同对照组进行比较,各组差异有统计学意义(x2=12.063,P0.05)。1.3 Atg5在急性淋巴细胞白血病同急性髓细胞白血病比较,两者表达差异无统计学意义(x2=0.090,P0.0125)。1.4 Atg5在急性淋巴细胞白血病同对照组比较,两者表达差异有统计学意义(x2=10.042,P0.0125)。1.5 Atg5在急性髓细胞白血病同对照组比较,两者表达差异有统计学意义(x2=6.667,P0.0125)。2 Atg7蛋白的表达情况2.1 Atg7在急性白血病中阳性率54.0%,在对照组阳性率30.0%,两者表达差异无统计学意义(x2=3.302,P0.05)。2.2 Atg7在急性淋巴细胞白血病中阳性率43.3%,在急性髓细胞白血病中阳性率70.0%,分别同对照组进行比较,各组差异有统计学意义(x2=6.7278,P0.05)。2.3 Atg7在急性淋巴细胞白血病同急性髓细胞白血病比较,两者间表达水平无明显差异(x2=3.435,P0.0125)。2.4 Atg7在急性淋巴细胞白血病同对照组比较,两者间表达水平无明显差异(x2=0.905,P0.0125)。2.5 Atg7在急性髓细胞白血病同对照组比较,两者表达差异有统计学意义(x2=6.400,P0.0125)。3利用Spearman秩相关性检验比较Atg5及Atg7蛋白表达是否相关,求得r=0.333,P0.01,两者在急性白血病中的表达无明显相关性。结论1 Atg5蛋白在急性淋巴细胞白血病及急性髓细胞白血病中表达上调,Atg5可能参与急性白血病的发生发展;Atg7蛋白在急性髓细胞白血病中表达上调,自噬活性增高,提示Atg7可能与急性髓细胞白血病的发生、发展有关。2实验发现Atg5、Atg7在急性白血病及非恶性血液病中均有表达,但其表达水平受疾病种类、分型的影响。3 Atg5、Atg7虽然都参与自噬体形成过程,但是两者在急性白血病中的表达无明显相关性。
[Abstract]:Objective: acute leukemia (acute leukemia AL) is a kind of common malignant hematopoietic stem cell proliferative diseases, the autophagy dysfunction is related to tumorigenesis. Autophagy is eukaryotic cells in autophagy related genes (autophagy related, gene, Atg) under the regulation and process of damaged organelles and macromolecules with the lysosomal degradation plays an important role in maintaining cell stability, is a basic regulation that cells adapt to internal and external environment. Immunohistochemical staining of bone marrow cells autophagy related gene Atg5 in children with acute leukemia, the expression level of Atg7 protein was detected on the two factor activity and childhood acute leukemia occurrence, development the relationship between the 1 subjects. Methods: in the experimental group were randomly selected from newly diagnosed pediatric hospital of Hebei Medical University second hospital from January 2014 to June 2016 in untreated acute leukemia 50 cases of children with acute lymphoblastic leukemia group 30 cases, 20 cases of acute myeloid leukemia group, control group and experimental group were selected to match the age and sex of non malignant hematological diseases (including immune thrombocytopenia, anaemia) in 20 children with.2 were collected and processed separately from: experimental group and control group fresh bone marrow specimens of 2ml, coagulation after 4% paraformaldehyde, embedded in paraffin, sliced.3 experimental methods: immunohistochemistry staining in children with acute lymphoblastic leukemia, acute myeloid leukemia and control group Atg5 in bone marrow cells, the expression level of.4 protein was Atg7 analysis: using SPSS13.0 statistical analysis software statistical analysis, Atg5, Atg7 protein expression in the specimens in strength using x2 test or Fisher exact test, P0.05 had statistical significance; Atg5, using the correlation of protein Atg7 Spearman correlation analysis, P0.01 was statistically significant. The expression of Atg5 1 protein 1.1 Atg5 in acute leukemia positive rate (82%) higher than that of the control group (positive rate 40%), there is a significant difference between the two (x2=12.0, P0.05).1.2 Atg5 in acute lymphoblastic leukemia positive rate was 83.3% in acute myeloid leukemia with positive rate of 80%, respectively, compared with the control group, the difference was statistically significant (x2=12.063, P0.05).1.3 Atg5 in acute lymphoblastic leukemia with acute myelogenous leukemia compared the expression difference was not statistically significant (x2=0.090, P0.0125).1.4 Atg5 in acute lymphoblastic leukemia compared with the control group, the expression of statistically significant difference (x2=10.042, P0.0125).1.5 Atg5 in acute myeloid leukemia compared with the control group, there were statistical significance of difference (x2=6.66 7, P0.0125) expression of.2 Atg7 protein 2.1 Atg7 in acute leukemia and the positive rate was 54%, the positive rate was 30% in the control group, the expression of the difference was not statistically significant (x2=3.302, P0.05).2.2 Atg7 in acute lymphoblastic leukemia with positive rate of 43.3%, in acute myeloid leukemia with positive rate of 70%, respectively, with the control groups were compared, the difference was statistically significant (x2=6.7278, P0.05).2.3 Atg7 in acute lymphoblastic leukemia compared with acute myeloid leukemia, there was no difference between the expression level of.2.4 Atg7 (x2=3.435, P0.0125) in acute lymphoblastic leukemia compared with the control group, no significant difference between the two levels (x2=0.905, P0.0125).2.5 Atg7 in acute myelogenous leukemia compared with the control group, the expression difference was statistically significant (x2=6.400, P0.0125).3 by the Spearman rank correlation test between the Atg5 and Atg7 protein expression As if, obtained r=0.333, P0.01, there was no significant correlation between their expression in acute leukemia. Conclusion the expression of Atg5 protein in 1 patients with acute lymphoblastic leukemia and acute myeloid leukemia up-regulated Atg5 may be involved in the development of acute leukemia; expression of Atg7 protein in acute myeloid leukemia up-regulated autophagy activity was increased, suggesting that Atg7 may be associated with acute myeloid leukemia, the development of the.2 experiments found that Atg5, the expression of Atg7 in acute leukemia and non malignant hematological diseases were, but its expression level by disease type,.3 Atg5 type Atg7 effects, although in autophagosome formation, but their expression in acute leukemia has no obvious correlation.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.71

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