發(fā)布時間：2018-04-09 16:22

  本文選題：白血病　切入點：髓細胞　出處：《吉林大學》2015年碩士論文

【摘要】：背景：慢性髓細胞白血�。–ML）簡稱慢粒，為惡性血液系統(tǒng)疾病，發(fā)生于骨髓造血干細胞，病程分為慢性期（CP）、加速期（AP）、最終急變期（BP/BC）。加速期、急變期被認為是疾病進展期，本病中位發(fā)病年齡在60歲~65歲。目前酪氨酸激酶抑制劑（TKI）成為慢粒治療的一線用藥，傳統(tǒng)觀點認為慢�；颊哌M入急性期首選異基因造血干細胞移植，且為可能治愈本病的唯一手段。否則病程僅為3~6個月，但移植費用昂貴且存在風險。所以臨床中治療以維持患者處于慢性期不進展為佳，需重點關注慢�；颊呒膊『螘r發(fā)生進展，但目前并沒有明確的分子標志物提示慢粒急變。研究顯示腫瘤的發(fā)生發(fā)展同表觀遺傳學密切相關，其中DNA甲基化是表觀遺傳調(diào)控的重要機制，甲基化異常是白血病發(fā)生過程中的一個普遍現(xiàn)象。研究表明在各種白血病中發(fā)現(xiàn)了多個基因高甲基化狀態(tài)，提示DNA甲基化異常修飾同白血病發(fā)病相關。前期研究工作顯示，人緊密連接蛋白（ZO-1）基因啟動子區(qū)，在急性白血病中呈現(xiàn)特異性高甲基化狀態(tài)，，與急性白血病發(fā)生、發(fā)展及轉(zhuǎn)歸密切相關，是白血病預后一個不良因素。 目的：對慢粒和急性白血病的細胞系以及慢�；颊卟〕滩煌瑫r期的骨髓標本，進行ZO-1基因啟動子區(qū)甲基化狀態(tài)檢測，探討慢粒疾病不同時期ZO-1基因啟動子區(qū)甲基化狀態(tài)的差異，及其與疾病分期轉(zhuǎn)變之間的關系。 方法：應用甲基化特異性聚合酶鏈反應（MS-PCR）及亞硫酸氫鈉聯(lián)合限制性內(nèi)切酶分析法（COBRA）檢測K562細胞及HL-60、Molt4等細胞，ZO-1基因啟動子區(qū)甲基化狀態(tài)，ZO-1基因的表達情況，以及去甲基化藥物對ZO-1基因甲基化狀態(tài)及表達的影響。檢測CML不同疾病分期骨髓標本ZO-1甲基化狀態(tài)，并通過擴大臨床標本數(shù)目，探討不同分期CML患者中ZO-1基因甲基化陽性患者比例。并對一例CML患者在不同分期階段骨髓標本檢測，檢測其慢性期、急變期、急變期治療后回到慢性期時ZO-1基因啟動子區(qū)甲基化狀態(tài)及表達的變化。 結(jié)果： 1. ZO-1基因啟動子區(qū)在K562細胞系中呈現(xiàn)非甲基化狀態(tài)，在HL-60、Molt-4細胞系中呈現(xiàn)特異性高甲基化狀態(tài)，同時在急性白血病細胞系中ZO-1表達沉默。經(jīng)去甲基化藥物地西他濱處理后的HL-60、Molt-4細胞系，ZO-1基因啟動子區(qū)由高甲基化狀態(tài)轉(zhuǎn)化為非甲基化狀態(tài)，同時恢復ZO-1基因的表達。 2. ZO-1基因啟動子區(qū)在正常人群呈現(xiàn)非甲基化狀態(tài)，慢粒患者慢性期呈現(xiàn)非甲基化狀態(tài)，慢�；颊呒铀倨诩凹弊兤谥谐尸F(xiàn)特異性高甲基化狀態(tài)。對10例CML-CP患者檢測，ZO-1基因均為非甲基化狀態(tài)，對8例CML-AP患者檢測，4例（50%）呈現(xiàn)高甲基化狀態(tài)，CML-BC患者7例，5例（70%）呈現(xiàn)ZO-1基因高甲基化狀態(tài)。 3.針對1例慢�；颊咴诼云诔尸F(xiàn)ZO-1基因非甲基化，急變期呈現(xiàn)高甲基化狀態(tài)，恢復至慢性期后甲基化狀態(tài)可轉(zhuǎn)變回非甲基化狀態(tài)。 結(jié)論： ZO-1基因啟動子區(qū)甲基化狀態(tài)同慢粒疾病不同時期有相關性。隨著疾病進展，ZO-1基因甲基化比例升高，出現(xiàn)高甲基化狀態(tài)提示可能有疾病進展傾向。
[Abstract]:Background: chronic myeloid leukemia (CML) CML, malignant hematological diseases, bone marrow hematopoietic stem cells, the course is divided into chronic phase (CP), accelerated phase (AP), the final blast phase (BP/BC). The acceleration period is considered in blastic phase progression of the disease, the disease incidence in at the age of 60 years. At present, ~65 tyrosine kinase inhibitors (TKI) become the first-line treatment of CML, the traditional view is that in acute phase of CML patients preferred allogeneic hematopoietic stem cell transplantation, and is the only possible means to cure the disease. It is only 3~ duration of 6 months, but the cost is high and there is a risk of transplant. So the clinical treatment of patients in chronic period not to maintain progress is good, need to focus on disease progression when CML patients, but there is no clear molecular marker for leukemia. Research shows that the occurrence and development of tumors with epigenetic density Closely related, in which DNA methylation is an important epigenetic regulation mechanism of table, abnormal methylation of leukemia is a common phenomenon in the process. The results show that multiple gene hypermethylation was found in all kinds of leukemia, suggesting that DNA methylation modification with leukemia disease. Previous study showed that people the tight junction protein (ZO-1) gene promoter region showed high specificity of methylation status in acute leukemia, the occurrence and development of acute leukemia, and the prognosis is closely related to the prognosis of leukemia is a bad factor.
Objective: to chronic and acute leukemia cell lines and the course of disease in patients with CML bone marrow specimens of different periods, promoter methylation status of ZO-1 gene promoter of detection, difference of CML in different periods of ZO-1 gene promoter methylation status, and the relationship between the change and staging of the disease.
Methods: methylation specific polymerase chain reaction (MS-PCR) and Sodium Bisulfite joint restriction enzyme analysis (COBRA) detection of K562 cells and HL-60 Molt4 cells, ZO-1 gene promoter methylation and expression of ZO-1 gene, and the effects of demethylation drugs on the expression and methylation status of ZO-1 gene detection of CML in different stages of disease. Bone marrow specimens of ZO-1 methylation, and by expanding the number of clinical specimens, explore the different stages of ZO-1 patients with CML gene methylation in patients with positive proportion. And on a case of CML patients in different stages of bone marrow specimens detection, detection of the chronic phase, acute phase, acute phase after treatment to chronic period the ZO-1 gene promoter and expression of promoter methylation status.
Result錛

本文編號：1727208