LncRNA SNHG7通过抑制miR-186增强人急性髓系白血病细胞耐药性

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (2): 171-177.

LncRNA SNHG7通过抑制miR-186增强人急性髓系白血病细胞耐药性

朱娟¹, 周英², 李颖佳^1*

1.中南大学湘雅三医院检验科, 湖南长沙 410013;
2.长沙市中心医院检验科, 湖南长沙 410004

收稿日期:2020-05-06 修回日期:2020-10-13 出版日期:2021-02-05 发布日期:2021-01-19
通讯作者: ^*37811309@qq.com
基金资助:
湖南省卫生健康委科研计划(20201453); 湖南省自然科学基金青年基金(2020JJ5877)

LncRNA SNHG7 enhances the drug resistance of human acute myeloid leukemia cells through inhibiting miR-186

ZHU Juan¹, ZHOU Ying², LI Ying-jia^1*

1. Department of Laboratory Medicine, the Third Xiangya Hospital of Central South University, Changsha 410013;
2. Department of Clinical Laboratory,Changsha Central Hospital,Changsha 410004, China

Received:2020-05-06 Revised:2020-10-13 Online:2021-02-05 Published:2021-01-19
Contact: ^*37811309@qq.com

摘要/Abstract

摘要： 目的探讨长链非编码RNA(lncRNA)SNHG7通过抑制miR-186对人急性髓系白血病(AML)细胞化疗耐药性的影响。方法取AML初诊患者、复发/难治患者及健康对照者的外周血;AML阿霉素敏感细胞(HL60)和AML阿霉素耐药细胞(HL60/ADM),采用荧光定量PCR检测外周血及细胞样本中SNHG7和miR-186的表达。CCK8法实验检测HL60和HL60/ADM细胞对阿霉素的敏感性并计算半数抑制浓度(IC₅₀)。在HL60/ADM细胞中转染sh-SNHG7、miR-186 mimic或miR-186 inhibitor,荧光定量PCR检测SNHG7和miR-186的表达,CCK8检测HL60/ADM对阿霉素的剂量反应和IC₅₀值。双荧光素酶报告基因实验检测SNHG7与miR-186的靶向结合。结果 AML初诊者和复发/难治者血清中SNHG7的表达水平显著高于对照组,miR-186的趋势与SNHG7相反(P＜0.01)。用不同浓度阿霉素处理后,HL60/ADM细胞活力显著高于HL60细胞(P＜0.05),且阿霉素对HL60/ADM细胞的IC₅₀值3.36±0.65显著高于HL60细胞0.43±0.16(P＜0.001)。HL60/ADM细胞中SNHG7显著高表达(P＜0.05),miR-186显著低表达(P＜0.05)。sh-SNHG7或miR-186 mimic转染HL60/ADM细胞后显著增加对阿霉素的敏感性,使IC₅₀值分别从3.17±0.61减少到2.30±0.31,3.22±0.62减少到2.16±0.33(P＜0.05)。双荧光素酶报告基因实验证实SNHG7的吸附靶基因为miR-186。结论 SNHG7通过抑制miRNA-186增强AML细胞化疗耐药性。

关键词: SNHG7, miR-186, 增殖, 耐药, 急性髓性白血病

Abstract: Objective To investigate the effect of long non-coding RNA (lncRNA) SNHG7 on cell drug resistance through inhibiting microRNA-186(miR-186). Methods Peripheral blood was taken from newly diagnosed AML patients, recurrent/refractory AML patients and healthy controls. AML adriamycin sensitive cells (HL60) and AML adriamycin resistant cells (HL60/ADM) were used to detect the expression of SNHG7 and miR-186 in peripheral blood and cell samples by fluorescence quantitative PCR. The sensitivity of HL60 and HL60/ADM cells to adriamycin was determined by CCK8 assay and the 50% inhibitory concentration(IC₅₀) was calculated. Sh-SNHG7 or miR-186 mimic or miR-186 inhibitor was transfected into HL60/ADM cells. The expres- sion of SNHG7 and miR-186 was detected by fluorescence quantitative PCR, the dose response and IC₅₀ of HL60/ADM to adriamycin were detected by CCK8. Double luciferase reporter gene assay was used to detect the targeted binding of SNHG7 to miR-186. Results The expression of SNHG7 in the serum of newly diagnosed AML and recurrent/refractory AML patients was significantly higher than that of normal controls, and the trend of miR-186 was contrary to that of SNHG7 (P＜0.01). Compared with HL60 cells, HL60/ADM cells were treated with adriamycin at different concentrations, and the IC₅₀ value of adriamycin on HL60/ADM (3.36±0.65) cells was significantly higher than that of HL60 cells (0.43±0.16) (P＜0.001). SNHG7 expression was significantly higher(P＜0.05) in HL60/ADM cells and miR-186 was significantly lower (P＜0.05). Transfection with sh-SNHG7 or miR-186 mimic in HL60/ADM cells increased the sensitivity to adriamycin and decreased the IC₅₀ value from 3.17±0.61 to 2.30±0.31 and 3.22±0.62 to 2.16±0.33, respectively (P＜0.05). Dual-luciferase reporter assay confirmed that SNHG7 could bind miR-186. Conclusions SNHG7 enhances drug resistance of AML cells during chemotherapy through inhibition of miR-186.

Key words: SNHG7, miR-186, proliferation, drug-resistance, acute myeloid leukemia

中图分类号:

R733.71

朱娟, 周英, 李颖佳. LncRNA SNHG7通过抑制miR-186增强人急性髓系白血病细胞耐药性[J]. 基础医学与临床, 2021, 41(2): 171-177.

ZHU Juan, ZHOU Ying, LI Ying-jia. LncRNA SNHG7 enhances the drug resistance of human acute myeloid leukemia cells through inhibiting miR-186[J]. Basic & Clinical Medicine, 2021, 41(2): 171-177.

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