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河北医学  2022, Vol. 28 Issue (11): 1768-1773    DOI: 10.3969/j.issn.1006-6233.2022.11.02
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CircBIRC6调节miR-126-5p/JARID2轴对非小细胞肺癌细胞顺铂耐药性的影响
党乙, 张梁, 翟恒钰, 李文海
西安国际医学中心医院胸外科, 陕西 西安 710100
Effect of CircBIRC6 Regulation of miR-126-5p/JARID2 Axis on Cisplatin Resistance in Non-Small Cell Lung Cancer Cells
DANG Yi, ZHANG Liang, ZHAI Hengyu, et al
Xi'an International Medical Center Hospital, Shaanxi Xi'an 710100, China
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摘要 目的: 探讨环状RNA BIRC6(CircBIRC6)对非小细胞肺癌(NSCLC)细胞(A549)顺铂(DDP)耐药性的影响。方法: RT-qPCR法检测DDP耐药及敏感肺癌组织与细胞中CircBIRC6表达,体外培养人NSCLC细胞株A549及DDP耐药细胞A549/DDP,将A549/DDP细胞分为Control组、pcDNA组、pcBIRC6组、si-CircBIRC6组、si-NC组、si-BIRC6+inhibitor NC组,si-BIRC6+miR-126-5p inhibitor组。RT-qPCR检测CircBIRC6、miR-126-5p表达;CCK-8法检测细胞增殖及对DDP的IC50值;流式细胞术检测细胞凋亡;Western blot检测JARID2、Cyclin D1、P21、P-gp蛋白表达;双荧光素酶报告基因实验验证CircBIRC6、JARID2与miR-126-5p的靶向关系。结果: 与DDP敏感肺癌组织相比,CircBIRC6在DDP耐药肺癌组织中表达水平显著升高,与A549细胞相比,A549/DDP细胞中CircBIRC6表达水平及DDP对A549/DDP细胞的IC50值显著升高(P<0.05);双荧光素酶报告基因实验证实CircBIRC6、JARID2与miR-126-5p存在靶向关系。与pcDNA组相比,pcBIRC6组A549/DDP细胞增殖、CircBIRC6、JARID2、Cyclin D1与P-gp蛋白表达及IC50值显著升高,细胞凋亡率及miR-126-5p、P21蛋白表达水平显著降低(P<0.05);与si-NC组相比,si-BIRC6组细胞增殖、CircBIRC6、JARID2、Cyclin D1与P-gp蛋白表达及IC50值显著降低,细胞凋亡率及miR-126-5p、P21蛋白表达水平显著升高(P<0.05);下调miR-126-5p表达可逆转抑制CircBIRC6表达对A549/DDP细胞的增殖抑制作用和凋亡促进作用。结论: CircBIRC6在A549/DDP细胞中高表达,可通过靶向抑制miR-126-5p表达,上调JARID2表达,促进A549/DDP细胞对DDP的耐药性。
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党乙
张梁
翟恒钰
李文海
关键词 环状RNA BIRC6微小RNA-126-5p非小细胞肺癌顺 铂    
AbstractObjective: To investigate the influence of circular RNA BIRC6 (CircBIRC6) on cisplatin (DDP) resistance of non-small cell lung cancer (NSCLC) cells (A549).Methods: RT-qPCR was used to detect the expression of CircBIRC6 in DDP-resistant and sensitive lung cancer tissues and cells. Human NSCLC cell line A549 and DDP-resistant cells A549/DDP were cultured in vitro. A549/DDP cells were grouped into Control group, pcDNA group, pcBIRC6 group, si-CircBIRC6 group, si-NC group, si-BIRC6+inhibitor NC group, and si-BIRC6+miR-126-5p inhibitor group. RT-qPCR was applied to detect the expression of CircBIRC6 and miR-126-5p; CCK-8 assay was applied to detect cell proliferation and IC50 value for DDP; flow cytometry was applied to detect apoptosis; Western blot was applied to detect the protein expressions of JARID2, Cyclin D1, P21, and P-gp; dual-luciferase reporter gene assay was applied to verify the targeting relationship between CircBIRC6, JARID2 and miR-126-5p. Results: Compared with DDP-sensitive lung cancer tissues, the expression level of CircBIRC6 in DDP-resistant lung cancer tissues was greatly higher; compared with A549 cells, the expression level of CircBIRC6 in A549/DDP cells and the IC50 value of DDP on A549/DDP cells were greatly higher (P<0.05); the dual-luciferase reporter gene assay confirmed that CircBIRC6 and JARID2 had a targeting relationship with miR-126-5p. Compared with the pcDNA group, the A549/DDP cell proliferation, the protein expressions of CircBIRC6, JARID2, Cyclin D1 and P-gp, and IC50 value in the pcBIRC6 group were greatly higher, the apoptosis rate and the expression levels of miR-126-5p and P21 proteins were greatly lower (P<0.05); compared with the si-NC group, the A549/DDP cell proliferation, the protein expressions of CircBIRC6, JARID2, Cyclin D1 and P-gp, and IC50 value in the si-BIRC6 group were greatly lower, the apoptosis rate and the expression levels of miR-126-5p and P21 proteins were greatly higher (P<0.05); down-regulation of miR-126-5p expression reversed the proliferation-inhibiting and apoptosis-promoting effects of inhibiting CircBIRC6 expression on A549/DDP cells. Conclusion: CircBIRC6 is highly expressed in A549/DDP cells, which can inhibit the expression of miR-126-5p by targeting, up-regulate the expression of JARID2, and promote the resistance of A549/DDP cells to DDP.
Key wordsCircular RNA BIRC6    microRNA-126-5p    Non-small cell lung cancer    Cisplatin
    
基金资助:陕西省重点研发计划项目,(编号:S2022-YF-YBSF-0779)
通讯作者: 张梁   
引用本文:   
党乙, 张梁, 翟恒钰, 李文海. CircBIRC6调节miR-126-5p/JARID2轴对非小细胞肺癌细胞顺铂耐药性的影响[J]. 河北医学, 2022, 28(11): 1768-1773.
DANG Yi, ZHANG Liang, ZHAI Hengyu, et al. Effect of CircBIRC6 Regulation of miR-126-5p/JARID2 Axis on Cisplatin Resistance in Non-Small Cell Lung Cancer Cells. HeBei Med, 2022, 28(11): 1768-1773.
链接本文:  
http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2022.11.02     或     http://www.hbyxzzs.cn/CN/Y2022/V28/I11/1768
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