目的验证急性早幼粒细胞白血病(APL)患者血肿瘤细胞中带核定位信号的维甲酸受体α(nuclear localization signal-retinoic acid receptor alpha,NLS-RARα)蛋白的存在及定位。方法采用蛋白质印迹法验证患者血肿瘤细胞中性粒细胞弹性蛋白酶(neutrophil elastase,NE)的存在;提取患者血肿瘤细胞核蛋白,蛋白质印迹法检测细胞核中NLS-RARα蛋白的表达;FITC/DAPI双染色免疫荧光法检测患者血肿瘤细胞中NLS-RARα的表达及定位;FITC/PI双染色激光共聚焦法检测患者血肿瘤细胞中NLS-RARα的表达及定位。以重组腺病毒Ad-NE感染的NB4细胞中NLS-RARα蛋白的表达及定位作阳性对照,以正常人血中性粒细胞中野生型RARα的表达和定位作阴性对照。结果阳性对照组设置成功。APL患者血肿瘤细胞中存在NE且有NLS-RARα蛋白表达。细胞免疫荧光法、激光共聚焦法检测结果提示APL患者血肿瘤细胞NLSRARα蛋白的表达主要位于胞核。结论成功用3种方法检测出APL患者血肿瘤细胞中NLS-RARα蛋白的存在并推测其定位,为进一步研究APL的早期诊断及复发监测提供了新思路。 Objective To verify the existence and localization of the nuclear localization signal-retinal acid receptor alpha (NLS-RARα) protein with nuclear localization signal in the blood of patients with acute promyelocytic leukemia (APL). Methods The presence of neutrophil elastase (NE) was detected by Western blotting. The expression of NLS-RARα protein in nuclei was detected by Western blotting. The FITC / DAPI double staining The expression and localization of NLS-RARα in the hematological tumor cells were detected by immunofluorescence. The expression and localization of NLS-RARα in the hematological tumor cells were detected by FITC / PI double staining confocal laser scanning microscope. The expression and localization of NLS-RARα protein in NB4 cells infected with recombinant adenovirus Ad-NE were used as positive control. The expression and localization of wild-type RARα in normal human neutrophils were used as negative control. Results The positive control group was set up successfully. There are NEs and NLS-RARα protein expression in the blood tumor cells of APL patients. Cell immunofluorescence and laser confocal microscopy showed that the expression of NLSRARα protein in blood cells of APL patients was mainly located in the nucleus. Conclusion The detection of the presence of NLS-RARα protein in blood tumor cells of APL patients by three different methods and their localization are inferred. It provides a new idea for the further study of APL early diagnosis and recurrence monitoring.