miR-520c-3p/CXCL8信号轴在慢性髓系白血病细胞红系分化中的调控作用

doi:10.3969/j.issn.1004-616x.2024.06.003

癌变·畸变·突变 ›› 2024, Vol. 36 ›› Issue (6): 438-443.doi: 10.3969/j.issn.1004-616x.2024.06.003

• 论著 • 上一篇

miR-520c-3p/CXCL8信号轴在慢性髓系白血病细胞红系分化中的调控作用

杨玉¹, 商宇¹, 倪蕾², 宫梓琳¹, 王然³, 王丹⁴, 王静艳²

1. 佳木斯大学基础医学院微生态-免疫调节网络与相关疾病重点实验室, 黑龙江佳木斯 154007;
2. 佳木斯大学附属第一医院, 黑龙江佳木斯 154003;
3. 佳木斯市中心医院, 黑龙江佳木斯 154002;
4. 佳木斯大学临床医学院, 黑龙江佳木斯 154003

收稿日期:2024-08-21 修回日期:2024-10-21 发布日期:2024-12-04
通讯作者: 王静艳
作者简介:杨玉,E-mail:yangyu@jmsu.edu.cn。
基金资助:
佳木斯大学青年创新人才培养支持计划项目(JMSUQP 2020012);黑龙江省卫生健康委科研课题(20210202040066)

Regulatory role of the miR-520c-3p/ CXCL8 axis in erythroid differentiation of chronic myeloid leukemia cells

YANG Yu¹, SHANG Yu¹, NI Lei², GONG Zilin¹, WANG Ran³, WANG Dan⁴, WANG Jingyan²

1. Key Laboratory of Microecology-Immune Regulatory Network and Related Diseases, Basic Medical College of Jiamusi University, Jiamusi 154007;
2. The First Affiliated Hospital of Jiamusi University, Jiamusi 154003;
3. Jiamusi Central Hospital, Jiamusi 154002;
4. Clinical Medical College of Jiamusi University, Jiamusi 154003, Heilongjiang, China

Received:2024-08-21 Revised:2024-10-21 Published:2024-12-04

摘要/Abstract

摘要： 目的： 探讨miR-520c-3p/CXCL8信号轴在慢性髓系白血病(CML)细胞红系分化中的作用及其机制。方法： 用氯化血红素诱导人CML细胞系K562细胞红系分化，逆转录实时荧光定量PCR(RT-qPCR)法检测CXCL8 mRNA的表达水平。用不同浓度CXCL8(0、20、40、60、80和100 ng/mL)处理K562细胞72 h，RT-qPCR实验检测γ-globin mRNA的表达情况，据此分析CXCL8对K562细胞红系分化的影响。将miR-520c-3p模拟物和inhibitor转染到K562细胞以增强或抑制miR-520c-3p的功能，RT-qPCR法检测γ-globin mRNA的表达水平，分析miR-520c-3p对K562细胞红系分化的影响。分别采用Western blot和RT-qPCR实验检测CXCL8 mRNA和蛋白的表达水平，观察miR-520c-3p对CXCL8表达的影响。细胞分为实验组(共转染miR-520c-3p模拟物与CXCL8 3'-UTR)和对照组(共转染模拟物对照与CXCL8 3'-UTR)，用双荧光素酶报告基因实验检测miR-520c-3p是否靶向CXCL8的3'-UTR。结果： 与对照组相比，氯化血红素诱导分化处理后，K562细胞中CXCL8 mRNA的表达水平升高(P<0.01)。与未处理组相比，不同剂量CXCL8处理组K562细胞中γ-globin mRNA的表达水平均明显升高(P<0.01)。与阴性对照组细胞相比，miR-520c-3p模拟物转染组K562细胞中γ-globin mRNA的表达水平、CXCL8 mRNA和蛋白表达水平均下降(均为P<0.01)；miR-520c-3p抑制物转染组细胞中γ-globin mRNA表达水平、CXCL8 mRNA和蛋白表达水平均上升(均为P<0.01)。双荧光素酶报告基因实验检测发现，与对照组相比，实验组的相对荧光素酶活性明显下降(P<0.01)。结论： 本研究揭示了miR-520c-3p/CXCL8信号轴在CML细胞红系分化中的关键作用。miR-520c-3p通过靶向调控CXCL8的表达，在K562细胞的红系分化过程中发挥抑制效应。

关键词: 慢性髓系白血病, miR-520c-3p, CXCL8, 红系分化

Abstract: OBJECTIVE： To explore the role and mechanism of the miR-520c-3p/CXCL8 axis in erythroid differentiation of chronic myeloid leukemia (CML). METHODS： K562 cells were induced to differentiate with hemin and treated with CXCL8 (0，20，40，60，80 and 100 ng/mL) for 72 h. Expression of γ-globin mRNA was examined by RT-qPCR to analyze the effect of CXCL8 on erythroid differentiation. miR-520c-3p mimics and inhibitor were transfected into K562 cells to enhance or inhibit miR-520c-3p function，respectively. Expression of γ-globin mRNA was measured by RT-qPCR to assess the impact of miR-520c-3p on erythroid differentiation of K562 cells. Western blot and RT-qPCR experiments were performed to detect CXCL8 expression and observe the influence of miR-520c-3p on CXCL8 expression. The cells were divided into the experiment group (co-transfected with miR-520c-3p mimics and CXCL8 3'-UTR) and the control group (co-transfected with mimics-control and CXCL8 3'-UTR)，a dual luciferase reporter gene assay was conducted to examine the targeting relationship between miR-520c-3p and the 3'-UTR of CXCL8. RESULTS： Compared to the control group，expression of CXCL8 mRNA was significantly increased after hemin-induced differentiation (P<0.01). Compared to the untreated group，the γ-globin mRNA expression of CXCL8 treatment groups were significantly elevated (P<0.01). Compared with the negative control group，the expression levels of γ-globin mRNA，CXCL8 mRNA，and CXCL8 protein in K562 cells transfected with miR-520c-3p mimics were all decreased (P<0.01). In cells transfected with the miR-520c-3p inhibitor，expression levels of γ-globin mRNA，CXCL8 mRNA，and CXCL8 protein were all increased (P<0.01). The dual luciferase reporter gene assay revealed a significant decrease in relative fluorescence activities in the experimental group compared to the control group (P<0.01). CONCLUSION： This study reveals the key role of the miR-520c-3p/CXCL8 signaling axis in the erythroid differentiation of CML cells. miR-520c-3p exerted an inhibitory effect on the erythroid differentiation of K562 cells by targeting the expression of CXCL8.

Key words: chronic myeloid leukemia, miR-520c-3p, CXCL8, erythroid differentiation

中图分类号:

R733.7

杨玉, 商宇, 倪蕾, 宫梓琳, 王然, 王丹, 王静艳. miR-520c-3p/CXCL8信号轴在慢性髓系白血病细胞红系分化中的调控作用[J]. 癌变·畸变·突变, 2024, 36(6): 438-443.

YANG Yu, SHANG Yu, NI Lei, GONG Zilin, WANG Ran, WANG Dan, WANG Jingyan. Regulatory role of the miR-520c-3p/ CXCL8 axis in erythroid differentiation of chronic myeloid leukemia cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(6): 438-443.

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miR-520c-3p/CXCL8信号轴在慢性髓系白血病细胞红系分化中的调控作用

Regulatory role of the miR-520c-3p/ CXCL8 axis in erythroid differentiation of chronic myeloid leukemia cells

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