miR-515-5p靶向RING1增强NSCLC A549/DDP细胞顺铂敏感性分析  

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] Xu Rui;Qian Yong;Zhang Haiyan(Department of Respiratory and critical medicine, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China;Department of Oncology, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China;Daytime Ward of Oncology, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China)

机构地区：[1]安徽医科大学第一附属医院呼吸与重症医学科,安徽230022 [2]安徽医科大学第一附属医院肿瘤内科,安徽230022 [3]安徽医科大学第一附属医院肿瘤日间病房,安徽230022

出　　处：《中华肺部疾病杂志（电子版）》2021年第5期559-563,共5页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：安徽省自然科学基金项目(2008085MA13)。

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。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 c

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 

分 类 号：R734.2[医药卫生—肿瘤]