川北医学院学报2025，Vol.40Issue(10)：1225-1229,1248,6.DOI:10.3969/j.issn.1005-3697.2025.10.001
miR-139-3p靶向SPP1对肺癌A549细胞顺铂耐药的影响及其作用机制
Effect and mechanism of miR-139-3p targeting SPP1 on cisplatin resist-ance in A549 lung cancer cells
摘要
Abstract
Objective:To explore the effect and mechanism of miR-139-3p targeting secreted phosphoprotein 1(SPP1)on cis-diamminedichloroplatinum(DDP)in A549 lung cancer cells.Methods:A cisplatin(DDP)-resistant A549/DDP cell model was established via gradient concentration induction,validated by resistance index(IR=3.877)and IC50 determination.The re-sistant cells were divided into five groups:blank control,negative control,miR-139-3p mimic,SPP1 overexpression,and SPP1 knockdown.Multi-platform analyses were performed,including qPCR(miR-139-3p and SPP1 expression),Western blot(SPP1,p-AKT,and total AKT protein levels),CCK-8(proliferation and drug sensitivity),flow cytometry(apoptosis),and du-al-luciferase reporter assays(miR-139-3p/SPP1 interaction).Results:The cisplatin-resistant A549/DDP cells exhibited a signifi-cantly higher IC50 value compared to parental A549 cells(P<0.05),with a resistance index(IR)of 3.877.Quantitative PCR(qPCR)analysis revealed that miR-139-3p expression was markedly upregulated in the miR-139-3p mimic group versus the negative control group(P<0.05).Relative to the miR-139-3p mimic group,SPP1 mRNA levels were significantly elevated in the SPP1-overex-pression group and reduced in the SPP1-knockdown group(P<0.05).Western blotting confirmed corresponding changes in SPP1 pro-tein expression across these groups(P<0.05).CCK-8 assays demonstrated that miR-139-3p overexpression enhanced the proliferation inhibition rate and reduced the IC50 value compared to the negative control(P<0.05),whereas SPP1 overexpression reversed these effects,lowering the inhibition rate and increasing IC50(P<0.05).Compared with the miR-139-3p blank control,the luciferase activity of the wild-type SPP13'UTR luciferase reporter plasmid binding to miR-139-3p analog decreased(P<0.05),and the difference in the luciferase activity of the mutant SPP13'UTR luciferase reporter plasmid binding to miR-139-3p analog was not statistically significant(P>0.05).Compared with the negative control group,the protein expression level of p-AKT in miR-139-3p control group was signifi-cantly decreased(P<0.05),compared with the miR-139-3p control group and SPP1 knockdown group,the protein expression level of p-AKT in SPP1 overexpression group was significantly increased(P<0.05).Conclusion:Overexpression of miR-139-3p or knockdown of SPP1 could inhibit the proliferation of A549/DDP cells,enhance their sensitivity to cisplatin,and the mechanism may be related to the AKT signaling pathway.关键词
微小RNA-139-3p/分泌型磷蛋白1/肺癌/耐药性/细胞增殖Key words
Micro RNA-139-3p/Secretory phosphoprotein 1/Lung cancer/Drug resistance/Cell proliferation分类
医药卫生引用本文复制引用
张令坤,康世荣..miR-139-3p靶向SPP1对肺癌A549细胞顺铂耐药的影响及其作用机制[J].川北医学院学报,2025,40(10):1225-1229,1248,6.基金项目
内蒙古自治区卫生健康委医疗卫生科技计划项目(202201269) （202201269）
