芬太尼通过ILF3-AS1/miR-132对卵巢癌细胞生长和转移的影响  

Effects of fentanyl on ovarian cancer cell growth and metastasis via ILF3-AS1/miR-132

作　　者：何含 刘群[1] 钟庆[1] 翁艳[1] 汪艳 黄凡 邬瑞刚 杨国仁[1] HE Han;LIU Qun;ZHONG Qing;WENG Yan;WANG Yan;HUANG Fan;WU Ruigang;YANG Guoren(Department of Anesthesiology and Pain,Jianyang People's Hospital,Jianyang,Sichuan 641400,China)

机构地区：[1]简阳市人民医院麻醉疼痛科,四川简阳641400

出　　处：《安徽医药》2022年第4期765-769,I0002,共6页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨芬太尼对卵巢癌细胞生长和转移的影响及分子机制。方法2018年8月至2019年10月,体外培养卵巢癌细胞A2780,用浓度分别为0、0.5、5、50、500 ng/mL的芬太尼处理,作为不同浓度芬太尼处理组。将过表达的白细胞介素增强结合因子3反义RNA1(ILF3-AS1)(pcDNA3.1-ILF3-AS1、对照pcDNA3.1)、抑制微小RNA(miRNA/miR)-132表达(抗-miR-132、对照anti-miR-NC)的载体分别转染A2780细胞,并以500 ng/mL芬太尼处理,记为芬太尼500+pcDNA3.1-ILF3-AS1组、芬太尼500+pcDNA3.1组、芬太尼500+anti-miR-132组、芬太尼500+anti-miR-NC组。将pcDNA3.1、pcDNA3.1-ILF3-AS1、si-NC、si-ILF3-AS1转染至A2780细胞中,记为pcDNA3.1组、pcDNA3.1-ILF3-AS1组、si-NC组、si-ILF3-AS1组。四甲基偶氮唑盐比色法(MTT)检测细胞存活率;克隆形成实验检测细胞克隆形成数;Transwell检测细胞迁移和侵袭;实时荧光定量PCR(RT-qPCR)检测ILF3-AS1和miR-132的表达水平;荧光素酶报告实验检测ILF3-AS1和miR-132的靶向关系。结果与0 ng/mL芬太尼处理组相比,5、50、500 ng/mL的芬太尼处理的A2780中miR-132表达水平显著升高[(1.39±0.13)、(1.68±0.17)、(2.34±0.23)比(1.00±0.10)],细胞存活率[(86.14±8.25)%、(71.03±7.11)%、(43.26±4.34)%比(100.01±10.01)%]、克隆形成数、迁移、侵袭细胞数显著降低,ILF3-AS1表达水平显著降低[(0.78±0.08)、(0.54±0.05)、(0.30±0.03)比(1.01±0.10)](P<0.05)。过表达ILF3-AS1和抑制miR-132表达均逆转了芬太尼对A2780增殖、迁移、侵袭的抑制作用。且ILF3-AS1靶向调控miR-132的表达。结论芬太尼浓度大于5 ng/mL时可抑制卵巢癌细胞增殖、迁移和侵袭,其机制可能与ILF3-AS1及miR-132有关。Objective To investigate the effect and molecular mechanism of fentanyl on the growth and metastasis of ovarian cancer cells.Methods From August 2018 to October 2019,ovarian cancer A2780 cells were cultured in vitro and treated with fentanyl at con⁃centrations of 0,0.5,5,50,and 500 ng/mL,as different fentanyl treatment groups.The interleukin enhancer binding factor 3 antisense RNA1(ILF3-AS1)overexpression plasmid(pcDNA3.1-ILF3-AS1,control pcDNA3.1)and microRNA-132(miR-132)siRNA(anti-miR-132,control anti-miR-NC vector)were transfected into A2780 cells and then treated with 500 ng/mL fentanyl,recorded as the fentanyl 500+pcDNA3.1-ILF3-AS1 group,fentanyl 500+pcDNA3.1 group,fentanyl 500+anti-miR-132 group and fentanyl 500+anti-miR-NC group.The pcDNA3.1,pcDNA3.1-ILF3-AS1,si-NC,si-ILF3-AS1 were transfected into A2780 cells and recorded as the pcDNA3.1 group,pcDNA3.1-ILF3-AS1 group,si-NC group and si-ILF3-AS1 group.Cell viability was detected by MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)assay;clone formation assay was used to detect the number of cell clones;Transwell was used to de⁃tect cell migration and invasion;real-time quantitative PCR(RT-qPCR)was used to detect ILF3-AS1 and miR-132 expression level;the targeting relationship between ILF3-AS1 and miR-132 was detected by luciferase reporter assay.Results Compared with the 0 ng/mL fentanyl-treatment group,the expression level of miR-132 in A2780 cells was significantly increased in the 5,50,and 500 ng/mL fentan⁃yl treatment groups[(1.39±0.13),(1.68±0.17),(2.34±0.23)vs(1.00±0.10)];the cell viability[(86.14±8.25)%,(71.03±7.11)%,(43.26±4.34)%vs(100.01±10.01)%],the number of colonies formed,the number of migrating and invasive cells was significantly decreased,and the expression level of ILF3-AS1 was significantly decreased[(0.78±0.08),(0.54±0.05),(0.30±0.03)vs(1.01±0.10)](P<0.05).Both ILF3-AS1 overexpression and miR-132 inhibition reversed the inhibitory effects of fentanyl on the proliferation,migration,and invasion of A2780 ce

关 键 词：芬太尼 白细胞介素增强结合因子3反义RNA1 微小RNA-132 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]