發(fā)布時(shí)間：2018-01-19 08:48

  本文關(guān)鍵詞： 急性白血病 自噬 免疫組織化學(xué)染色 Atg5 Atg7 蛋白　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:急性白血病(acute leukemia,AL)是一類常見的造血干細(xì)胞惡性增殖性疾病,目前認(rèn)為自噬調(diào)控失調(diào)與腫瘤發(fā)生密切相關(guān)。自噬是真核細(xì)胞在自噬相關(guān)基因(autophagy related gene,Atg)的調(diào)控下利用溶酶體降解受損細(xì)胞器和大分子物質(zhì)的過程,在維持細(xì)胞穩(wěn)定方面起重要作用,是細(xì)胞適應(yīng)內(nèi)外環(huán)境所需的一種基本調(diào)節(jié)方式。采用免疫組織化學(xué)染色法對(duì)兒童急性白血病骨髓細(xì)胞中自噬相關(guān)基因Atg5、Atg7蛋白表達(dá)水平進(jìn)行檢測,探討這兩種因子活性與兒童急性白血病發(fā)生、發(fā)展的關(guān)系。方法1研究對(duì)象:實(shí)驗(yàn)組隨機(jī)選取自2014年1月至2016年6月河北醫(yī)科大學(xué)第二醫(yī)院兒科住院的初診未治急性白血病患兒50例,其中急性淋巴細(xì)胞白血病組30例,急性髓細(xì)胞白血病組20例,對(duì)照組選取同期與實(shí)驗(yàn)組年齡性別相匹配的非惡性血液病(包括免疫性血小板減少癥、營養(yǎng)性貧血)患兒20例。2標(biāo)本采集和處理:分別抽取實(shí)驗(yàn)組及對(duì)照組新鮮骨髓標(biāo)本2ml,自然凝固后經(jīng)4%多聚甲醛固定,常規(guī)石蠟包埋、切片。3實(shí)驗(yàn)方法:采用免疫組織化學(xué)染色法檢測兒童急性淋巴細(xì)胞白血病、急性髓細(xì)胞白血病及對(duì)照組骨髓細(xì)胞中Atg5、Atg7蛋白的表達(dá)水平。4統(tǒng)計(jì)學(xué)分析:應(yīng)用SPSS13.0統(tǒng)計(jì)分析軟件進(jìn)行統(tǒng)計(jì)學(xué)分析,Atg5、Atg7蛋白在各組標(biāo)本中表達(dá)強(qiáng)度采用x2檢驗(yàn)或Fisher確切概率法,P0.05有統(tǒng)計(jì)學(xué)意義;Atg5、Atg7蛋白的相關(guān)性使用Spearman相關(guān)性分析進(jìn)行檢驗(yàn),P0.01有統(tǒng)計(jì)學(xué)意義。結(jié)果1 Atg5蛋白的表達(dá)情況1.1 Atg5在急性白血病中陽性率(82.0%)高于對(duì)照組陽性率(40.0%),兩者之間表達(dá)差異有統(tǒng)計(jì)學(xué)意義(x2=12.0,P0.05)。1.2 Atg5在急性淋巴細(xì)胞白血病中陽性率為83.3%,在急性髓細(xì)胞白血病中陽性率80.0%,分別同對(duì)照組進(jìn)行比較,各組差異有統(tǒng)計(jì)學(xué)意義(x2=12.063,P0.05)。1.3 Atg5在急性淋巴細(xì)胞白血病同急性髓細(xì)胞白血病比較,兩者表達(dá)差異無統(tǒng)計(jì)學(xué)意義(x2=0.090,P0.0125)。1.4 Atg5在急性淋巴細(xì)胞白血病同對(duì)照組比較,兩者表達(dá)差異有統(tǒng)計(jì)學(xué)意義(x2=10.042,P0.0125)。1.5 Atg5在急性髓細(xì)胞白血病同對(duì)照組比較,兩者表達(dá)差異有統(tǒng)計(jì)學(xué)意義(x2=6.667,P0.0125)。2 Atg7蛋白的表達(dá)情況2.1 Atg7在急性白血病中陽性率54.0%,在對(duì)照組陽性率30.0%,兩者表達(dá)差異無統(tǒng)計(jì)學(xué)意義(x2=3.302,P0.05)。2.2 Atg7在急性淋巴細(xì)胞白血病中陽性率43.3%,在急性髓細(xì)胞白血病中陽性率70.0%,分別同對(duì)照組進(jìn)行比較,各組差異有統(tǒng)計(jì)學(xué)意義(x2=6.7278,P0.05)。2.3 Atg7在急性淋巴細(xì)胞白血病同急性髓細(xì)胞白血病比較,兩者間表達(dá)水平無明顯差異(x2=3.435,P0.0125)。2.4 Atg7在急性淋巴細(xì)胞白血病同對(duì)照組比較,兩者間表達(dá)水平無明顯差異(x2=0.905,P0.0125)。2.5 Atg7在急性髓細(xì)胞白血病同對(duì)照組比較,兩者表達(dá)差異有統(tǒng)計(jì)學(xué)意義(x2=6.400,P0.0125)。3利用Spearman秩相關(guān)性檢驗(yàn)比較Atg5及Atg7蛋白表達(dá)是否相關(guān),求得r=0.333,P0.01,兩者在急性白血病中的表達(dá)無明顯相關(guān)性。結(jié)論1 Atg5蛋白在急性淋巴細(xì)胞白血病及急性髓細(xì)胞白血病中表達(dá)上調(diào),Atg5可能參與急性白血病的發(fā)生發(fā)展;Atg7蛋白在急性髓細(xì)胞白血病中表達(dá)上調(diào),自噬活性增高,提示Atg7可能與急性髓細(xì)胞白血病的發(fā)生、發(fā)展有關(guān)。2實(shí)驗(yàn)發(fā)現(xiàn)Atg5、Atg7在急性白血病及非惡性血液病中均有表達(dá),但其表達(dá)水平受疾病種類、分型的影響。3 Atg5、Atg7雖然都參與自噬體形成過程,但是兩者在急性白血病中的表達(dá)無明顯相關(guān)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.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R733.71

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