摘要
目的探讨瑞芬太尼对肾癌(RCC)细胞生物学行为的影响和机制。方法通过四甲基偶氮唑蓝(MTT)法、克隆形成实验、Transwell实验、实时荧光定量PCR(RT-qPCR)检测不同浓度的瑞芬太尼对RCC细胞786-0增殖、克隆迁移与侵袭,以迁移及基因间长链非编码RNA00462(Linc00462)、微RNA(miR)-2355-5p表达的影响。将Linc00462小干扰RNA(si-Linc00462)、Linc00462过表达质粒(pcDNA-Linc00462)分别转染786-0细胞,观察干扰Linc00462或过表达Linc00462对786-0细胞生物学行为的影响。双荧光素酶报告实验和RT-qPCR分析Linc00462和miR-2355-5p的靶向关系。结果瑞芬太尼处理显著降低786-0细胞活力、克隆形成数、迁移数和侵袭数(P<0.05),降低Linc00462表达水平(P<0.05),增加miR-2355-5p表达水平(P<0.05)。干扰Linc00462表达显著降低786-0细胞克隆形成数、迁移数和侵袭数(P<0.05),增加miR-2355-5p表达水平(P<0.05)。过表达Linc00462显著增加786-0克隆形成数、迁移数和侵袭数(P<0.05),降低miR-2355-5p表达水平(P<0.05)。过表达Linc00462显著减弱瑞芬太尼处理对786-0细胞增殖、迁移和侵袭的抑制作用(P<0.05)。双荧光素酶报告实验结果显示Linc00462能与miR-2355-5p直接结合。结论瑞芬太尼可抑制RCC细胞增殖、迁移和侵袭,其机制可能与下调Linc00462/miR-2355-5p通路表达有关。
Objective To investigate the effect and mechanism of remifentanil on the biological behavior of renal cancer(RCC)cells.Methods The effects of different concentrations of remifentanil on the proliferation,cloning,migration,invasion and long intergenic non-coding RNA(Linc00462)and microRNA(miR)-2355-5p expression of RCC cells 786-0 were detected by the methyl thiazolyl tetrazolium(MTT)method,clone formation assay,Transwell assay,real-time quantitative PCR(RT-qPCR).Linc00462 small interfering RNA(si-Linc00462),Linc00462 overexpression plasmid(pcDNA-Linc00462)were respectively transfected into 786-0 cells to observe the effect of interference with Linc00462 or Linc00462 overexpression on the biological behavior.The dual luciferase report assay and RT-qPCR were applied to analyze targeting relationship between Linc00462 and miR-2355-5p.Results Remifentanil treatment significantly reduced the viability of 786-0 cells,clonal formation number,migration number and invasion number of 786-0 cells(P<0.05),decreased the expression level of Linc00462(P<0.05),and increased the expression level of miR-2355-5p(P<0.05)in 786-0 cells.Interference with Linc00462 expression significantly reduced clonal formation number,migration and invasion number of 786-0 cells(P<0.05),and increased the expression level of miR-2355-5p(P<0.05).Overexpression level of Linc00462 significantly increased clonal formation number,migration and invasion number of 786-0(P<0.05),and decreased the expression of miR-2355-5p(P<0.05).Overexpression of Linc00462 significantly reduced the inhibitory effects of remifentanil on the proliferation,migration and invasion of 786-0 cells(P<0.05).The results of the dual luciferase report assay showed that Linc00462 could directly bind to miR-2355-5p.Conclusion Remifentanil can inhibit the proliferation,migration and invasion of RCC cells,and its mechanism may be related to the down-regulation of Linc00462/miR-2355-5p axis.
作者
文竹
李军
何洁
WEN Zhu;LI Jun;HE Jie(Department of Anesthesiology,Mianyang Central Hospital/Affiliated Mianyang Hospital,School of Medicine,University of Electronic Science and Technology of China,Mianyang,Sichuan 621000,China)
出处
《重庆医学》
CAS
2022年第7期1093-1099,共7页
Chongqing medicine
基金
四川省重点实验室研究基金2020年度项目(SYS20-01)。
作者简介
文竹(1983-),副主任医师,本科,主要从事超声在围术期中应用研究;通信作者:李军,E-mail:lj89199@163.com。
