miR-515-5p靶向SPHK1调控K562细胞的红系分化研究

doi:10.3969/j.issn.1671-7171.2019.11.018

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Abstract 【Objective】To investigate whether miR-515-5p targets SPHK1 to regulate erythroid differentiation in K562 cells. 【Methods】TargetScanHuman was used to analyze the match status between miR-515-5p and SPHK1. To detect miR-515-5p targeting SPHK1, the luciferase reporter system was applied. In the case of miR-515-5p mimics overexpression or miR-515-5p inhibitor knock-down of miR-7-5p, the expression levels of erythroid differentiation-related genes HBE, HBG1, HBB, GATA1, and KLF1 were measured by qPCR, and the expression levels of erythroid differentiation marker proteins CD71 and CD235a were detected by WB. 【Results】The luciferase reporter experiment showed that miR-515-5p targeted the 3'UTR of SPHK1. When miR-515-5p was overexpressed, the expression levels of erythroid differentiation-related genes HBE, HBG1, HBB, GATA1 and KLF1 in K562 cells decreased (P<0.05). The expression of SPHK1 decreased at both d0 and d₄ (P<0.05).While the expression levels of the erythroid differentiation marker proteins CD71 and CD235a increased at d₄ (P<0.05). Compared to the control group, the expression levels of CD71 and CD235a were lower (P<0.05). On the other hand, when knocking down miR-515-5p, the expression levels of erythroid differentiation related genes HBE, HBG1, HBB, GATA1 and KLF1 in K562 cells increased (P<0.05) and the expression of SPHK1 increased at d0 and d₄ (P<0.05). The expression levels of the erythroid differentiation marker proteins CD71 and CD235a increased at d₄ as well (P<0.05), however, on the 4th day the expression levels of CD71 and CD235a were higher compared to the control group (P<0.05). 【Conclusion】miR-515-5p inhibits the erythroid differentiation regulation of K562 cells after its targeting SPHK1.

Key words： Erythropoiesis/IM MicroRNAs Gene Expression Regulation

Received: 20 May 2019

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R555.1

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http://journal07.magtech.org.cn/yxlcyj/EN/10.3969/j.issn.1671-7171.2019.11.018 OR http://journal07.magtech.org.cn/yxlcyj/EN/Y2019/V36/I11/2137

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