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Difference in glycerol levels between leukemia and normal bone marrow stem cells

机译:白血病和正常骨髓干细胞之间甘油水平的差异

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Aquaglyceroporin 9 (AQP9) is considered to be involved in numerous types of carcinogenic processes, particularly in liver carcinoma. AQP9 expression is significantly decreased in the human hepatocellular carcinoma when compared with the non-tumourigenic liver, which leads to increased resistance to apoptosis. In addition, AQP9 is permeable to glycerol and urea. The involvement of AQP9 in leukemia has not been fully delineated. It is proposed that abnormal proliferation of hematopoietic stem cells (HSCs) contributes to leukemia carcinogenesis. Therefore, the present study aimed to investigate the possible roles of AQP9 in HSCs and its effect on the intracellular glycerol content. HSCs and non-HSCs (nHSCs) were isolated via magnetic-activated cell sorting and then subjected to flow cytometry for evaluation of purity. White blood cells (WBCs) were isolated from peripheral blood from healthy volunteers. Furthermore, AQP9 expression was examined at the mRNA and protein levels using western blotting and reverse transcription-polymerase chain reaction (RT-PCR). The glycerol content of HSCs, nHSCs and WBCs was evaluated by ELISA. Finally, in order to observe the morphology of HSCs and nHSCs, a blood smear was conducted and the cells were observed with Wright-Giemsa staining. The results indicated that the glycerol content in the HSCs was markedly greater than that in the nHSCs. AQP9 mRNA and protein expression was not detected in the HSCs and nHSCs, but was identified in the WBCs. Moreover, the HSC morphological characteristics included round or oval cells with round, slightly oval or irregularly shaped nuclei. Additionally, the nuclei occupied almost the entire cell, were located in the middle or were biased toward one side, and were stained light purple or red. Overall, 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 in further understanding the mechanism of leukemia carcinogenesis, and these data may be valuable in developing future therapeutic strategies.
机译:水生甘油9(AQP9)被认为与多种致癌过程有关,特别是在肝癌中。与非致瘤性肝相比,人肝细胞癌中AQP9的表达明显降低,这导致对细胞凋亡的抵抗力增强。此外,AQP9可以渗透甘油和尿素。尚未完全确定AQP9与白血病的关系。有人提出造血干细胞(HSCs)的异常增殖有助于白血病的癌变。因此,本研究旨在调查AQP9在HSC中的可能作用及其对细胞内甘油含量的影响。通过磁激活细胞分选法分离HSC和非HSC(nHSC),然后进行流式细胞术以评估纯度。从健康志愿者的外周血中分离出白细胞(WBC)。此外,使用蛋白质印迹法和逆转录聚合酶链反应(RT-PCR)在mRNA和蛋白水平检测AQP9表达。通过ELISA评估HSC,nHSC和WBC的甘油含量。最后,为了观察HSC和nHSC的形态,进行了血液涂片检查并通过Wright-Giemsa染色观察了细胞。结果表明,HSC中的甘油含量明显高于nHSC中的甘油含量。在HSC和nHSC中未检测到AQP9 mRNA和蛋白表达,但在WBC中已鉴定出AQP9 mRNA和蛋白表达。此外,HSC的形态学特征包括圆形或椭圆形细胞,具有圆形,略呈椭圆形或不规则形状的核。另外,核几乎占据了整个细胞,位于中间或偏向一侧,并被染成淡紫色或红色。总体而言,我们的结果表明,尽管甘油不是由AQP9介导的,但细胞内甘油与HSC增殖有关。因此,我们的发现可能有助于进一步了解白血病的致癌机制,并且这些数据可能对制定未来的治疗策略具有重要意义。

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