浙江医学2025，Vol.47Issue(12)：1240-1247,1252,9.DOI:10.12056/j.issn.1006-2785.2025.47.12.2024-1254
lncRNA NEAT1靶向miR-379-3p调控自噬在肺腺癌培美曲塞化疗耐药性中的作用研究
LncRNA NEAT1 targets miR-379-3p to regulate autophagy in pemetrexed chemotherapy resistance in lung adenocarcinoma
摘要
Abstract
Objective To investigate the mechanism of long noncoding RNA(lncRNA)nuclear paraspeckle assembly transcript 1(NEAT1)targeting miR-379-3p to regulate autophagy in pemetrexed(PEM)chemotherapy resistance in lung adenocarcinoma(LAD).Methods The expression of lncRNA NEAT1 and miR-379-3p in LAD cells was detected by quantitative real-time polymerase chain reaction.Two PEM drug-resistant LAD cell A549/PEM and NCI-H1299/PEM were established,and shRNA that interferes with lncRNA NEAT1 expression(shNEAT1)and miR-379-3p inhibitor were constructed and transfected into A549/PEM and NCI-H1299/PEM cells.Cell counting kit-8 and clonal formation assays were used to detect cell viability and proliferation.The rate of apoptosis was detected by flow cytometry.Western blot and immunofluorescence were used to detect the autophagy.Dual luciferase reporter gene assay was used to detect the targeting binding relationship between lncRNA NEAT1 and miR-379-3p.Results Compared with normal bronchial epithelial cells 16HBE,the expression of lncRNA NEAT1 was significantly elevated in the LAD cells,while the expression of miR-379-3p was significantly decreased in the LAD cells.lncRNA NEAT1 was further up-regulated in the PEM-resistant LAD cells,and the expression of miR-379-3p was further down-regulated.The addition of autophagy inhibitors to PEM-resistant LAD cells inhibited autophagy and apoptosis,and enhanced cell activity.The addition of autophagy agonists,on the other hand,promoted cellular autophagy and apoptosis,and reduced cell activity.After A549/PEM and NCI-H1299/PEM were transfected with shNEAT1,cell activity and cell proliferation were significantly inhibited,the expression of autophagy proteins microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ and Beclin-1 were significantly up-regulated,while p62 was significantly down-regulated.NEAT1 bound to miR-379-3p and suppressed its expression.Transfection of miR-379-3p inhibitor in A549/PEM and NCI-H1299/PEM cells reversed the expression of autophagy proteins induced by shNEAT1,as well as the attenuation of cell activity and proliferation.The effects of shNEAT1 on cellular autophagy as well as cell activity and proliferation were further reversed when mammalian target of rapamycin(mTOR)agonists were added.Conclusion lncRNA NEAT1 can regulate mTOR pathway by targeting miR-379-3p to inhibit autophagy of LAD cells,and thus enhance LAD cells' resistance to PEM,and this mechanism can provide a reliable theoretical basis for future targeted therapy against LAD.关键词
长链非编码RNA核旁斑组装转录本1/微小RNA-379-3p/肺腺癌/自噬/化疗耐药性Key words
LncRNA NEAT1/miR-379-3p/Lung adenocarcinoma/Autophagy/Chemotherapy resistance引用本文复制引用
何梦,孙健,陶克龙,唐任绮..lncRNA NEAT1靶向miR-379-3p调控自噬在肺腺癌培美曲塞化疗耐药性中的作用研究[J].浙江医学,2025,47(12):1240-1247,1252,9.基金项目
浙江省医药卫生科技计划面上项目(2020KY322) （2020KY322）
绍兴市卫生健康科技计划项目(2022KY006) （2022KY006）
绍兴市基础公益类计划项目(2023A14006) （2023A14006）
