首页|LncRNA MAGI2-AS3通过靶向调控miR-1269a/PTEN/AKT通路增强非小细胞肺癌对顺铂化疗的敏感性

LncRNA MAGI2-AS3通过靶向调控miR-1269a/PTEN/AKT通路增强非小细胞肺癌对顺铂化疗的敏感性

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目的 研究lncRNA MAGI2-AS3对非小细胞肺癌顺铂耐药的影响及其分子机制。方法 采用qRT-PCR检测MAGI2-AS3和miR-1269a在顺铂敏感细胞(A549，H1299)和顺铂耐药细胞(A549/DDP，H1299/DDP)中的表达差异。采用慢病毒敲低A549和H1299中的MAGI2-AS3的表达，过表达A549/DDP和H1299/DDP中的MAGI2-AS3并加入20 μmol/L顺铂(DDP)处理。A549/DDP和H1299/DDP细胞实验分组为:OE-NC组，OE-MAGI2-AS3组，OE-NC+DDP组，OE-MAGI2-AS3+DDP组，A549和H1299细胞实验分组为:sh-NC组，sh-MAGI2-AS3组，sh-NC+DDP组，sh-MAGI2-AS3+DDP组。CCK-8和细胞克隆形成实验检测细胞存活率，流式细胞术和Western blotting分别检测细胞凋亡率和蛋白表达量，划痕实验和Transwell检测细胞EMT变化。通过网站GEPIA、StarBase和miRDB预测MAGI2-AS3、miR-1269a和PTEN之间的靶向关系，并采用荧光素酶报告基因实验和RIP实验验证。采用miR-1269a mimic和pcDNA3。1-PTEN进行Rescue实验。结果 MAGI2-AS3在肺癌组织中的表达显著低于正常癌旁组织(P<0。05)且与患者不良预后相关(P<0。05)。A549/DDP和H1299/DDP中的MAGI2-AS3表达量显著低于A549和H1299(P<0。01)。干扰MAGI2-AS3可以显著促进顺铂处理前、后A549和H1299细胞的存活及EMT进程(P<0。01)，同时降低顺铂诱导的细胞凋亡(P<0。005)。过表达MAGI2-AS3可以显著抑制顺铂处理前、后A549/DDP和H1299/DDP细胞存活与EMT进程(P<0。01)，同时提升顺铂诱导的细胞凋亡(P<0。005)。MAGI2-AS3与miR-1269a结合在一起，miR-1269a与PTEN 3'UTR结合。在A549/DDP细胞内MAGI2-AS3通过靶向吸附miR-1269a促进PTEN的表达并下调AKT磷酸化从而抑制顺铂刺激下细胞的EMT进程(P<0。01)、促进顺铂诱导的A549/DDP细胞凋亡(P<0。01)。结论 LncRNA MAGI2-AS3通过靶向调控miR-1269a/PTEN/AKT信号轴增强非小细胞肺癌对顺铂化疗的敏感性。
LncRNA MAGI2-AS3 enhances cisplatin sensitivity of non-small cell lung cancer cells by regulating the miR-1269a/PTEN/AKT pathway
Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

long noncoding RNA MAGI2-AS3miR-1269anon-small cell lung cancercisplatin resistance

范喜瑞、戚之琳、邓园洁、杨子晗、孙丽、李国豪、梁娟娟、吴菲、袁力文

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皖南医学院 基础医学院生物化学与分子生物学教研室,安徽 芜湖 241002

皖南医学院 药学院,安徽 芜湖 241002

皖南医学院 临床医学院,安徽 芜湖 241002

皖南医学院 基础医学院生理学教研室,安徽 芜湖 241002

皖南医学院 检验学院,安徽 芜湖 241002

皖南医学院 麻醉学院,安徽 芜湖 241002

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LncRNA MAGI2-AS3 miR-1269a 非小细胞肺癌 顺铂耐药

安徽省高校自然科学研究重点项目安徽省高校自然科学研究重点项目安徽省高校自然科学研究重点项目安徽省高校自然科学研究重点项目皖南医学院自然科学重点科研项目皖南医学院自然科学重点科研项目皖南医学院自然科学重点科研项目大学生创新创业训练计划项目大学生创新创业训练计划项目

2022AH051212KJ2020ZD542023AH0517492022AH051243WK2021Z16WK2022Z03WK2022Z09202310368001S202310368030

2024

10.12122/j.issn.1673-4254.2024.10.22

南方医科大学学报
南方医科大学

南方医科大学学报

CSTPCD北大核心
影响因子：1.654
ISSN：1673-4254
年,卷(期)：2024.44(10)