【摘要】：目的:本研究旨在建立穩(wěn)定、有效、重復性好的人急性早幼粒白血病重度聯(lián)合免疫缺陷(SCID)小鼠移植瘤模型,觀察模型的病程特點和生物學行為。方法:將3-5周齡SCID beige小鼠隨機分為實驗組和對照組;實驗組鼠尾靜脈注射對數(shù)生長期的NB4細胞5×106個/只,動態(tài)監(jiān)測小鼠外周血白細胞數(shù)和外周血涂片中人早幼粒細胞陽性率;應用形態(tài)學和組織病理學方法確認白血病細胞浸潤肝、脾、肺、腎、腦等器官情況,Western blot檢測組織中PML-RARa融合蛋白表達情況。結果:接種NB4細胞2周內(nèi)的實驗組SCID小鼠外周血白細胞計數(shù)與對照組相比差異無統(tǒng)計學意義(P0.05);同時,血涂片姬姆薩染色后鏡檢未見到NB4細胞;接種第21天和第28天實驗組SCID小鼠外周血白細胞計數(shù)分別為(4.79±1.13)×109/L和(7.62±2.24)×109/L,顯著高于對照組(P0.05);血涂片吉姆薩染色后可見NB4細胞比例分別占(2.14±0.63)%和(6.6±2.76)%;形態(tài)學觀察顯示,接種21 d后實驗組SCID小鼠體內(nèi)出現(xiàn)單個或多個腫瘤實體包塊;同時,組織切片HE染色表明,肝、脾、肺、腎、腦組織均有不同程度的瘤細胞浸潤;細胞免疫熒光結果顯示,實驗組小鼠骨髓細胞CD33表達呈陽性(P0.05);Western blot數(shù)據(jù)證實,肝、腎、腦組織中檢測到不同水平的PML-RARa融合蛋白表達。結論:SCID小鼠鼠尾靜脈注射NB4細胞可成功構建人急性早幼粒細胞白血病小鼠模型,該模型能模擬臨床白血病累及骨髓和彌漫生長特點,是研究人類白血病發(fā)病機理及實驗治療的良好模型。
[Abstract]:Objective: to establish a stable, effective and reproducible human acute promyelocytic leukemia severe combined immunodeficient (SCID) mouse model, and to observe the course of disease and biological behavior of the model. Methods: SCID beige mice aged 3 to 5 weeks were randomly divided into experimental group and control group. In the experimental group, 5 脳 10 ~ 6 NB4 cells were injected intravenously into the tail vein to dynamically monitor the number of peripheral blood leukocytes and the positive rate of human progranulocytes in peripheral blood smears. The expression of PML-RARa fusion protein in leukemic cells infiltrated into liver, spleen, lung, kidney and brain was detected by morphology and histology., Western blot was used to detect the expression of PML-RARa fusion protein in liver, spleen, lung, kidney, brain and other organs. Results: there was no significant difference in peripheral blood leukocyte count between the experimental group and the control group within 2 weeks after inoculated with NB4 cells (P 0.05). At the same time, no NB4 cells were found in the blood smear after Giemsa staining. On the 21st and 28th day of vaccination, the peripheral blood leukocyte counts of SCID mice in the experimental group were (4.79 鹵1.13) 脳 109 脳 L and (7.62 鹵2.24) 脳 109 / L, respectively, which were significantly higher than those in the control group (P 0.05). The percentage of NB4 cells in blood smears was (2.14 鹵0.63)% and (6.6 鹵2.76)%, respectively, and the morphological observation showed that one or more tumor solid masses appeared in SCID mice in the experimental group 21 days after vaccination, and the percentage of DNA cells in the experimental group was (2.14 鹵0.63)% and (6.6 鹵2.76)%, respectively. At the same time, HE staining showed that there were different degrees of tumor cell infiltration in liver, spleen, lung, kidney and brain tissue, and the results of cellular immunofluorescence showed that the expression of CD33 in bone marrow cells of the experimental group was positive (P 0.05). Western blot data confirmed that different levels of PML-RARa fusion protein expression were detected in liver, kidney and brain tissue. Conclusion: the mouse model of human acute promyelocytic leukemia can be successfully established by injecting NB4 cells into the tail vein of SCID mice. The model can simulate the characteristics of clinical leukemia involving bone marrow and diffused growth. It is a good model to study the pathogenesis and experimental treatment of human leukemia.
【作者單位】： 西安交通大學醫(yī)學院第一附屬醫(yī)院血液內(nèi)科;
【基金】：陜西省科技統(tǒng)籌創(chuàng)新工程計劃項目(2012KTCL03-12)
【分類號】：R733.7;R-332

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