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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摘要 【目的】探讨miR-515-5p是否靶向SPHK1调控K562细胞的红系分化。【方法】通过TargetScanHuman网站分析miR-515-5p与SPHK1的匹配情况,然后通过荧光素酶报告系统检测miR-515-5p是否靶向SPHK1;在miR-515-5p mimics过表达或者miR-515-5p inhibitor敲低miR-7-5p的情况下,通过qPCR检测红系分化相关基因HBE、HBG1、HBB、GATA1、KLF1的表达量,通过WB检测红系分化标志蛋白CD71和CD235a的表达量。【结果】miR-515-5p靶向SPHK1的3'UTR;过表达miR-515-5p时,K562细胞的红系分化相关基因HBE、HBG1、HBB、GATA1、KLF1的表达量均下降(P<0.05),SPHK1的表达量在d₀和d₄均下降(P<0.05),红系分化标志蛋白CD71和CD235a的表达量在d₄均上升(P<0.05),但是d₄时CD71和CD235a的表达量相对于对照组减少(P<0.05);敲低miR-515-5p时,K562细胞的红系分化相关基因HBE、HBG1、HBB、GATA1、KLF1的表达量均上升(P<0.05),SPHK1的表达量在d₀和d₄均上升(P<0.05),红系分化标志蛋白CD71和CD235a的表达量在d₄均上升(P<0.05),但d₄时CD71和CD235a的表达量相对于对照组增多(P<0.05)。【结论】miR-515-5p靶向SPHK1后抑制调控K562细胞的红系分化。

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关键词 ：红细胞生成/免疫学, 微RNAs, 基因表达调控

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

收稿日期: 2019-05-20

PACS:

R555.1

引用本文:

廉诚, 姚远, 杨沛, 韩杨, 王长鹰. miR-515-5p靶向SPHK1调控K562细胞的红系分化研究[J]. 医学临床研究, 2019, 36(11): 2137-2140.
LIAN Cheng, YAO Yuan, YANG Pei, et al. miR-515-5p Targets SPHK1 to Regulate Erythroid Differentiation of K562 Cells. JOURNAL OF CLINICAL RESEARCH, 2019, 36(11): 2137-2140.

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