1Department of Oncology, Chinese PLA General Hospital, Beijing, 100853, China.
Department of Clinical Nutrition, Chinese PLA General Hospital, Beijing, 100853, China.
Sci Rep. 2018 Apr 4;8(1):5674. doi: 10.1038/s41598-018-23968-1.
This study is to explore the molecular regulation mechanism of CD133 which is associated with malignancy and poor prognosis of blood system diseases. CD133+HUCB-MNC (human umbilical cord blood mononuclear cells) and CD133-HUCB-MNC were isolated and amplificated from umbilical cord blood, and then were exposed to different doses of radiation and subjected to a clonogenic assay. CCK-8 kit was used to detect cell viability, Annexin V-FITC/PI cell apoptosis detection kit was used for the detection of apoptotic cells and the BrdU assay was performed by flow cytometry. The expression of protein was analyzed by western blots. The profile of miRNA expression in response to radiation was examined and validated by RT-PCR. miR-142-3p inhibited the expression of CD133 in umbilical cord blood mononuclear cells to increase radiosensitivity. CD133+HUCB-MNC cells were more radioresistant compared with CD133-HUCB-MNC cells. CD133+HUCB-MNC cells showed higher p-AKT and p-ERK levels after radiation. And miR-142-3p acted on 3'UTR of CD133 mRNA to inhibit CD133 expression. Moreover, miRNA-142-3p mimic increased radiosensitivity in CD133+HUCB-MNC cells. Our results elucidated a novel regulation pathway in hematopoietic stem cells and suggested a potential therapeutic approach for blood system diseases therapy.
本研究旨在探索与血液系统疾病恶性程度和预后不良相关的 CD133 的分子调控机制。从脐带血中分离和扩增 CD133+HUCB-MNC（人脐带血单个核细胞）和 CD133-HUCB-MNC，然后用不同剂量的辐射照射，并进行集落形成试验。CCK-8 试剂盒用于检测细胞活力，Annexin V-FITC/PI 细胞凋亡检测试剂盒用于检测凋亡细胞，BrdU 实验通过流式细胞术进行。通过 Western blot 分析蛋白表达。通过 RT-PCR 检查和验证 miRNA 表达谱对辐射的反应。miR-142-3p 抑制脐带血单个核细胞中 CD133 的表达，从而提高放射敏感性。与 CD133-HUCB-MNC 细胞相比，CD133+HUCB-MNC 细胞对辐射的抵抗力更强。CD133+HUCB-MNC 细胞在辐射后显示出更高的 p-AKT 和 p-ERK 水平。并且 miR-142-3p 作用于 CD133 mRNA 的 3'UTR 以抑制 CD133 表达。此外，miRNA-142-3p 模拟物增加了 CD133+HUCB-MNC 细胞的放射敏感性。我们的结果阐明了造血干细胞中的一个新的调控途径，并为血液系统疾病的治疗提供了一种潜在的治疗方法。
