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| 急性早幼粒细胞白血病患者血肿瘤细胞NLS-RARα蛋白的表达与定位 |
| 王慧1,2,刘北忠1,2,阳小群2,朱新瑜2,马鹏鹏2,蒋开玲2,钟梁2* |
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(1. 重庆医科大学附属永川医院中心实验室, 重庆 402160;
2. 重庆医科大学临床检验诊断学教育部重点实验室, 重庆 400016
*通信作者) |
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| 摘要: |
| 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NLS-RARα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。 |
| 关键词: 维甲酸受体α 核定位信号 中性粒细胞 急性早幼粒细胞白血病 |
| DOI：10.3724/SP.J.1008.2015.00241 |
| 投稿时间：2014-03-05修订日期：2014-08-06 |
| 基金项目:国家自然科学基金(81171658),重庆市自然科学基金重点项目(2011BA5037). |
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| Location of NLS-RARα protein in peripheral blood tumor cells of patients with acute promyeolic leukemia |
| WANG Hui1,2,LIU Bei-zhong1,2,YANG Xiao-qun2,ZHU Xin-yu2,MA Peng-peng2,JIANG Kai-ling2,ZHONG Liang2* |
(1. Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, China;
2. Key Laboratory of Clinical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China
*Corresponding author) |
| Abstract: |
| Objective To verify the presence and location of the NLS-RARα protein in the peripheral blood tumor cells of patients with acute promyeolic leukemia (APL). Methods Western blotting analysis was used to identify the NE enzyme in the peripheral blood tumor cells of APL patients. The nucleoprotein in tumor cells was prepared and NLS-RARα protein was detected by Western blotting analysis. The expression and location of NLS-RARα protein in peripheral blood tumor cells of APL patients were examined by FITC/DAPI double immunofluorescence staining and FITC/PI double staining laser confocal microscopy. The expression and location of NLS-RARα protein in NB4 cells infected with recombinant adenovirus Ad-NE was used as positive control and those of wildtype RARα in the neutrophils of healthy controls were taken as negative control. Results Positive control was successfully established. NE enzyme and NLS-RARα protein were expressed in peripheral blood tumor cells of APL patients. Immunofluorescence and laser confocal findings indicated that NLS-RARα protein was mainly located at the nuclei of peripheral blood tumors cells in APL patients. Conclusion NLS-RARα protein has been successfully detected by 3 different methods in the peripheral blood tumor cells of APL patients and its intracellular location has also been proposed, which can contribute to the early diagnosis and recurrence monitoring of APL. |
| Key words: retinoic acid receptor α nuclear localization signals neutrophils acute promyelocytic leukemia |
