[Effect of Hypoxia on CD47 Expression in Human Acute Myeloid Leukemia Cell Lines].

Chao-Yu Wang, Jie-Ping Li, Ying-Yu Nan, Qi-Ying Li, Ying Xiang, Yao Liu
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Abstract

Objective: To investigate the effect of hypoxia on hypoxia-inducible factor 1α (HIF-1α) and CD47 expression in human acute myeloid leukemia (AML) cell lines.

Methods: The CD47 expression in AML U937, HL-60, and K562 cells lines were detected by flow cytometry. U937, HL-60, and K562 cells were all divided into hypoxia-treated group and conventional oxygen group. The hypoxia-treated group was cultured with 1% O2, while the conventional oxygen group was cultured with 20% O2, then the cells were collected after 24 hours. Real time PCR was used to examine the mRNA changes of CD47 gene. Western blot assay was applied to detect the protein expression of HIF-1α and CD47.

Results: The expression of CD47 in U937, HL-60, and K562 cells was 98% (98%±0.03%), 99% (99%±0.05%), and 75% (75%±0.11%), respectively. The real time PCR showed that the mRNA expression of CD47 in U937 and HL-60 cells were up-regulated in the hypoxia-treated group (P<0.05), while in K562 cells was not (P>0.05). Western blot result showed that the protein levels of HIF-1α and CD47 of U937, HL-60, and K562 cells in the hypoxia-treated group were increased compared with the conventional oxygen group (P<0.05).

Conclusion: The hypoxia can up-regulate the expression of CD47 in acute myeloid leukemia cells, which may be related to HIF-1α.

缺氧对人急性髓系白血病细胞CD47表达的影响
目的:探讨缺氧对人急性髓性白血病(AML)细胞缺氧诱导因子1α (HIF-1α)和CD47表达的影响。方法:采用流式细胞术检测AML U937、HL-60和K562细胞系中CD47的表达。U937、HL-60、K562细胞均分为缺氧处理组和常规氧处理组。缺氧处理组以1% O2培养,常规氧组以20% O2培养,24h后收集细胞。采用Real - time PCR检测CD47基因mRNA的变化。Western blot法检测HIF-1α和CD47蛋白的表达。结果:CD47在U937、HL-60和K562细胞中的表达率分别为98%(98%±0.03%)、99%(99%±0.05%)和75%(75%±0.11%)。real - time PCR结果显示,缺氧组U937和HL-60细胞中CD47 mRNA表达上调(P0.05)。Western blot结果显示,缺氧处理组U937、HL-60和K562细胞中HIF-1α和CD47蛋白水平较常规氧处理组升高(结论:缺氧可上调急性髓系白血病细胞中CD47的表达,这可能与HIF-1α有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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