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目的:研究氧化砷(As2O3)和全反式维甲酸(AT-RA)对PLZF-RARα阳性细胞的作用。方法:稳定转染PLZF-RARα基因的U937细胞(U937/PLZF)经As2O3、ATRA处理后,Wright-Giemsa染色观察细胞形态,MTT法检测细胞生长增殖状态,流式细胞术(FCM)检测细胞周期和细胞表面分化抗原CD11b、CD64、CD14等的表达变化,荧光免疫细胞化学染色检测融合蛋白表达,细胞化学染色和硝基蓝四氮唑(NBT)还原试验观察细胞功能分化情况。结果:U937/PLZF细胞在去除四环素条件培养后,PLZF-RARα表达明显增加。As2O3(0.5μmol/L)联合ATRA(1μmol/L)使U937/PLZF细胞核质比缩小,核仁减少但未消失;生长增殖受抑;S期细胞减少;CD11b表达增高;PLZF-RARα蛋白表达减弱,分布以胞核弥漫细小颗粒为主。细胞化学染色和NBT反应变化不明显。结论:0.5μmol/L As2O3联合1μmol/L ATRA可使PLZF-RARα阳性U937细胞产生轻微的形态学分化趋势,尚不足以引起其发生功能分化。
Objective: To investigate the effects of As2O3 and AT-RA on PLZF-RARα-positive cells. METHODS: U937 cells (U937 / PLZF) stably transfected with PLZF-RARα gene were treated with As2O3 and ATRA, and the cell morphology was observed by Wright-Giemsa staining. Cell proliferation and proliferation were detected by MTT assay. Cell cycle was detected by flow cytometry (FCM) And cell surface differentiation antigen CD11b, CD64, CD14 and so on. Fluorescence immunocytochemistry was used to detect the expression of fusion protein. Cytochemical staining and NBT test were used to observe the differentiation of cells. Results: The expression of PLZF-RARα in U937 / PLZF cells was significantly increased after tetracycline-free culture. As2O3 (0.5μmol / L) combined with ATRA (1μmol / L) reduced the ratio of nucleus to cytoplasm of U937 / PLZF cells but did not disappear; the growth and proliferation were inhibited; the number of S phase cells decreased; the expression of CD11b increased; the expression of PLZF- , Distributed to the nucleus diffuse small particles. Cytochemical staining and NBT reaction did not change significantly. CONCLUSION: 0.5μmol / L As2O3 combined with 1μmol / L ATRA can induce slight morphological differentiation of U937 cells, which is still not enough to induce functional differentiation of PLZF-RARα positive U937 cells.