白血病干细胞中水通道蛋白1与水通道蛋白9表达的意义-东南大学学报医学版

白血病干细胞中水通道蛋白1与水通道蛋白9表达的意义

单位：徐州医科大学病理生理学教研室, 江苏徐州 221004

分类号：R733.7

出版年·卷·期（页码）：2019·38·第四期（712-717）

摘要：

目的：检测白血病干细胞中是否有水通道蛋白1（AQP1）与水通道蛋白9（AQP9）的表达，判断其与白血病细胞异常增殖间可能存在的关系。方法：采用免疫磁珠法（MACS）分离纯化白血病患者骨髓中CD34⁺CD38^-细胞，流式细胞术（FACS）检测分离纯化程度；Western blotting检测各组中AQP1与AQP9蛋白的表达；RT-PCR检测各组中AQP1与AQP9 mRNA的表达；甘油试剂盒检测各组中甘油含量。结果：纯化后CD34⁺CD38^-细胞占60.06%，CD34⁺CD38⁺细胞占35.15%，CD34^-CD38^-细胞占5.47%，CD34^-CD38⁺细胞占1.32%；AQP9 mRNA在外周血白细胞组中有表达；Western blotting结果显示AQP9蛋白在外周血白细胞组中有表达；甘油试剂盒检测显示各细胞均含有甘油。结论：细胞内甘油参与造血干细胞增殖，甘油不是由AQP9介导的，这有助于进一步了解白血病致癌机制。

Objective: In order to study the relationship between aquaporin 1(AQP1) and aquaporin 9(AQP9) and the abnormal proliferation of leukemia cells, we detected the expression of AQP1 and AQP9 protein and mRNA in leukemia stem cells. Methods: (1) CD34⁺CD38^- cells were isolated from leukemia patient bone marrow by using magnetic-activated cell sorting (MACS), and the purity rate of the sorted CD34⁺CD38^- cells was measured by flow cytometry (FACS); (2) The expression of AQP1 and AQP9 mRNA in every group cells was detected by RT-PCR; (3) The expression of AQP1 and AQP9 protein in every group cells was detected by Western blotting; (4) The content of glycerin in every group cells was detected by glycerol kit.Results: (1) The purity rate of the sorted CD34⁺CD38^- cells, CD34⁺CD38⁺ cells, CD34^-CD38^- cells and CD34^-CD38⁺ cells was 60.06%, 35.15%, 5.47%, 1.32%, respectively; (2) The result of Western blotting showed that the expression of AQP9 protein was only in positive control peripheral blood white blood cells group; (3) The expression of AQP9 mRNA was only in positive control peripheral blood white blood cells group; (4) The result of glycerol kit showed that every group cells contained glycerin.Conclusion: Our results indicated that intracellular glycerol is involved in HSC proliferation, despite the fact that glycerol is not mediated by AQP9. Hence, our findings may be useful for further understanding the mechanism of leukemia carcinogenesis.

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