2023 年第 9 期 第 18 卷

硒元素通过靶向长链非编码RNA核富集转录本1/微小RNA-149-5p信号轴来调节非小细胞肺癌细胞的顺铂耐药

Selenium affecting the cisplatin resistance phenotype of non-small cell lung cancer cells by regulating long non-coding RNA nuclear-enriched abundant transcript 1/microRNA-149-5p signal axis

作者：陈雯辉1郭晓彤2

英文作者：Chen Wenhui1 Guo Xiaotong2

单位：1首都医科大学附属北京天坛医院胸外科,北京100070；2中国医学科学院肿瘤医院深圳医院胸外科，深圳518116

英文单位：1Department of Thoracic Surgery Beijing Tiantan Hospital Capital Medical University Beijing 100070 China; 2Department of Thoracic Surgery Cancer Hospital Chinese Academy of Medical Sciences Shenzhen Center Shenzhen 518116 China

关键词：非小细胞肺癌；长链非编码RNA；微小RNA；顺铂耐药；硒供体

英文关键词：Non-smallcelllungcancer;Longnon-codingRNA;MicroRNA;Cisplatinresistance;Seleniumdonor

• 摘要：

• 目的  探讨硒元素调节非小细胞肺癌（NSCLC）细胞顺铂耐药的分子机制。方法 利用硒元素供体甲基亚硒酸（MSeA）以不同浓度联合顺铂处理商品化NSCLC细胞系A549及其顺铂耐药表型细胞系A549/DDP，并测定MSeA对细胞顺铂耐药表型和分子表达的影响。利用Lipofectamine2000转染试剂对细胞进行过表达质粒及干扰质粒的转染；利用实时荧光定量聚合酶链反应技术检测细胞中长链非编码RNA核富集转录本1（NEAT1）和微小RNA（miR）-149-5p的表达；检测细胞的增殖、凋亡及半胱氨酸天冬氨酸蛋白酶3（Caspase-3）活性，从而确定相关处理对肿瘤细胞的调节作用。结果 MSeA与顺铂共处理A549/DDP细胞（联合处理组）的细胞活力相对定量低于顺铂处理组，细胞凋亡率高于顺铂处理组［（30±13）%比（88±16）%、（23.6±8.5）%比（6.8±1.4）%］，差异均有统计学意义（均P<0.01）。MSeA共处理可部分恢复顺铂对A549的NEAT1表达的影响、联合处理组NEAT1相对定量低于顺铂处理组（P<0.001）；药物联合并NEAT1过表达质粒转染组A549和A549/DDP细胞 miR-149-5p相对定量高于联合处理组（P<0.01或P<0.05）。药物联合并NEAT1过表达质粒转染组、药物联合并miR-149-5p inhibitor转染组细胞活力相对定量高于联合处理组，细胞凋亡率、Caspase-3活性低于联合处理组（P<0.05或P<0.01），药物联合并miR-149-5p mimics转染组以上指标与联合处理组差异均无统计学意义（均P>0.05）。结论 硒元素可通过靶向长链非编码RNA NEAT1/微小RNA-149-5p信号轴来调节NSCLC细胞的顺铂耐药，MSeA具有抗顺铂耐药肿瘤的临床应用潜力。

• Objective To investigate the function of selenium on regulating non-small cell lung cancer(NSCLC) cells cisplatin resistance. Methods Commercialized NSCLC cell lines A549 and its cisplatin resistant pattern A549/DDP was used in this study. The regulatory effect of methylseleninic acid(MSeA), a selenium donor, on cisplatin resistance and molecular expression was determined by co-treatment of MSeA and cisplatin. Cells were transfected with plasmid using Lipofectamine2000 transfection reagent; the expressions of long non-coding RNA nuclear-enriched abundant transcript 1(NEAT1) and microRNA(miR)-149-5p were detected by quantitative real-time-polymerase chain reaction assay; cell proliferation, apoptosis and Caspase-3 activity were detected to determine the regulatory effect of relevant treatment on tumor cells. Results The relative quantification of cell viability of A549/DDP cells co-treated with MSeA and cisplatin(combined treatment group) was lower than that of cisplatin treatment group, and the apoptosis rate was higher than that of cisplatin treatment group［(30±13)% vs (88±16)%, (23.6±8.5)% vs (6.8±1.4)%］(both P<0.01). MSeA co-treatment can partially restore the effect of cisplatin on NEAT1 expression in A549, and the relative quantification of NEAT1 in the combined treatment group was lower than that in the cisplatin treatment group(P<0.001); the relative quantification of miR-149-5p in A549 and A549/DDP cells transfected with combined and NEAT1 overexpression plasmids transfection group was higher than that in the combined treatment group(P<0.01 or P<0.05). The cell viability of the combined and NEAT1 overexpression plasmid transfection group and combined and miR-149-5p inhibitor transfection group was relatively quantitatively higher than that of the combined treatment group, while the cell apoptosis rate and Caspase-3 activity were lower than those of the combined treatment group(P<0.05 or P<0.01); there was no statistically significant difference in the above indicators between the combined and miR-149-5p mimics transfection groups and the combined treatment group(all P>0.05). Conclusions  sSelenium can affect the cisplatin resistance phenotype of NSCLC cells by regulating long non-coding RNA NEAT1/miR-149-5p signal axis, MSeA has potential for clinical application against cisplatin resistant tumors.