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《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:

期数:

2021年05期

页码:

559-563

栏目:

论著

出版日期:

2021-10-20

文章信息/Info

Title:

Study on the mechanism of miR-515-5p enhanced the cisplatin sensitivity of NSCLC A549/DDP cells via targeting RING1

作者:

许 锐¹; 钱 勇²; 张海燕³

230022 安徽,安徽医科大学第一附属医院呼吸与重症医学科¹、肿瘤内科²、肿瘤日间病房³

Author(s):

Xu Rui¹;  Qian Yong²;  Zhang Haiyan³.

¹Department of Respiratory and critical medicine, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China; ²Department of Oncology, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China; ³Daytime Ward of Oncology, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China

关键词:

微小RNA-515-5p;  无名指蛋白1;  非小细胞肺癌;  顺铂敏感性

Keywords:

MicroRNA-515-5p;  Ring finger protein 1;  Non-small cell lung cancer;  Cisplatin sensitivity

分类号:

R734.2

DOI:

10.3877/cma.j.issn.1674-6902.2021.05.003

摘要:

目的 探讨微小RNA-515-5p(miR-515-5p)对非小细胞肺癌(non-small cell lung cancer, NSCLC)细胞顺铂敏感性的影响及机制。方法 实时荧光定量PCR(RT-qPCR)检测BEAS-2B、A549及A549/DDP细胞中miR-515-5p的表达情况。A549/DDP细胞分为NC组、miR-515-5p组、si-RING1组及miR-515-5p inh+si-RING1组,CCK-8法检测A549/DDP细胞增殖活性,用WB检测RING1和凋亡相关蛋白的表达情况。用双荧光素酶报告基因系统验证miR-515-5p与RING1的靶向关系。结果 miR-515-5p在BEAS-2B、A549及A549/DDP中的表达水平分别为(1.00±0.03)、(0.61±0.03)、(0.31±0.04); A549/DDP细胞转染miR-515-5p mimics后,NC组和miR-515-5p组细胞在24、48、72和96 h时的OD₄₅₀值分别为(0.61±0.02)、(0.52±0.02),(0.84±0.02)、(0.68±0.04),(1.03±0.03)、(0.79±0.04),(1.08±0.03)、(0.86±0.04); 以上指标组间差异均有统计学意义(P<0.05)。miR-515-5p组cleaved caspase-3、cleaved PARP和Bax表达水平显著高于NC组,Bcl-2的表达水平显著低于NC组。A549/DDP细胞转染si-RING1、miR-515-5p inhibitor后,NC组,si-RING1组和miR-515-5p inh+si-RING1组细胞在24、48、72和96 h时的OD₄₅₀值分别为(0.63±0.02)、(0.49±0.04)、(0.58±0.03)、(0.83±0.03)、(0.62±0.05)、(0.80±0.02)、(1.05±0.04)、(0.76±0.03)、(1.02±0.04)、(1.15±0.05)、(0.86±0.03)、(1.12±0.06); 以上指标组间差异均有统计学意义(P<0.05)。si-RING1组cleaved caspase-3、cleaved PARP和Bax表达水平显著高于NC组和miR-515-5p inh+si-RING1组,Bcl-2的表达水平显著低于NC组和miR-515-5p inh+si-RING1组。结论 miR-515-5p通过靶向下调RING1的表达增强A549/DDP细胞顺铂敏感性。

Abstract:

Objective To investigate the effect and mechanism of microRNA-515-5p(miR-515-5p)on the cisplatin sensitivity of non-small cell lung cancer(NSCLC). Methods The expression of miR-515-5p in BEAS-2B, A549 and A549/DDP cells was detected by real-time fluorescent quantitative PCR(RT-qPCR). A549/DDP cells were divided into NC group, miR-515-5p group, si-RING1 group and miR-515-5p inh+si-RING1 group. CCK-8 assay was used to assess the proliferation of A549/DDP cells. WB was performed to measure the expression of RING1 and apoptosis related proteins. Dual luciferase reporter gene system was used to verify the targeting relationship between miR-515-5p and RING1. Results The expression of miR-515-5p in BEAS-2B, A549 and A549/DDP cells were(1.00±0.03),(0.61±0.03),(0.31±0.04), respectively. A549/DDP cells were transfected with miR-515-5p mimics, the OD₄₅₀ values of cells in NC group and miR-515-5p group at 24, 48, 72 and 96 h were(0.61±0.02),(0.52±0.02),(0.84±0.02),(0.68±0.04),(1.03±0.03),(0.79±0.04),(1.08±0.03),(0.86±0.04), respectively. The differences of those items were all statistically significant(all P<0.05). The expression of cleaved caspase-3, cleaved PARP and Bax in miR-515-5p group were significantly higher than those in NC group, and the expression of Bcl-2 were significantly lower than those in NC group. A549/DDP cells were transfected with si-RING1 and miR-515-5p inhibitor, the OD₄₅₀ values of cells in NC group, si-RING1 group and miR-515-5p inh+si-RING1 group at 24, 48, 72 and 96 h were(0.63±0.02),(0.49±0.04),(0.58±0.03),(0.83±0.03),(0.62±0.05),(0.80±0.02),(1.05±0.04),(0.76±0.03),(1.02±0.04),(1.15±0.05),(0.86±0.03),(1.12±0.06), respectively. The differences of those items were all statistically significant(all P<0.05). The expression of cleaved caspase-3, cleaved PARP and Bax in si-RING1 group were significantly higher than those in NC group and miR-515-5p inh+siRING1 group, and the expression of Bcl-2 were significantly lower than those in NC group and miR-515-5p inh+siRING1 group. Conclusion miR-515-5p enhanced the cisplatin sensitivity of A549/DDP cells through targeting downregulate the expression of RING1.

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备注/Memo

备注/Memo:

基金项目: 安徽省自然科学基金项目(2008085MA13)

常用功能

更新日期/Last Update: 2021-10-20