血清细胞外纳米囊泡 miR-412-3p 靶向调控 TEAD1 促进亚厘米肺结节恶性生物学行为的机制研究。

Mechanism study of serum extracellular nano-vesicles miR-412-3p targeting regulation of TEAD1 in promoting malignant biological behavior of sub-centimeter lung nodules.

机构信息

Department of Oncology, Zhongda Hospital, Medical School, Southeast University, Nanjing, China.

Department of Cardiothoracic Surgery, Children's Hospital of Nanjing Medical University, Nanjing, China.

出版信息

Cancer Biomark. 2024;41(1):69-82. doi: 10.3233/CBM-240137.

DOI:10.3233/CBM-240137

PMID:39269825

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11495320/

Abstract

OBJECTIVE

To investigate the impact and potential mechanisms of serum extracellular nano-vesicles (sEVs) miR-412-3p released from sub-centimeter lung nodules with a diameter of ⩽ 10 mm on the malignant biological function of micro-nodular lung cancer (mnLC).

METHODS

A total of 87 participants were included and divided into a mnLC group (n= 30), a benign lung nodule (BLN) group (n= 27), and a healthy people control group (n= 30). Transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blot (WB) were used to measure the morphological characteristics and surface markers of sEVs. In vitro analysis, real-time quantitative polymerase chain reaction (RT-qPCR), CCK-8 cell proliferation assay, clone formation assay, Transwell, stem cell sphere-forming assay, and WB assay were conducted to verify the effect of miR-412-3p/TEAD1 signaling axis on the biological function of lung cancer cells through, respectively. Further validation was conducted using the serum sEVs of the participants.

RESULTS

The expression level of sEVs-miR-412-3p in the mnLC group was significantly higher than that in the BLN and healthy groups (P< 0.01). In lung cancer cell lines, miR-412-3p can negatively regulate the targeted gene TEAD1. The miR-412-3p/TEAD1 signaling axis is involved in promoting the EMT signaling pathway and regulating the malignant biological functions of lung cancer cell proliferation, migration, and stemness (P< 0.05). In addition, sEVs in the mnLC group significantly promoted lung cancer cell proliferation, migration, and stemness compared to the BLN and healthy groups, inhibited the expression of E-cadherin and TEAD1 in lung cancer cells, and promoted the expression of N-cadherin and Vimentin (P< 0.05).

CONCLUSION

sEVs-miR-412-3p could promote the biological process of EMT, and lead to the occurrence of malignant biological behavior in sub-centimeter lung nodules. This provides evidence for the miR-412-3p/TEAD1 signaling axis as a potential therapeutic target for mnLC.

摘要

目的

研究直径 ⩽ 10mm 的亚厘米肺结节中血清细胞外纳米囊泡（sEVs）miR-412-3p 对微小结节肺癌（mnLC）恶性生物学功能的影响及潜在机制。

方法

共纳入 87 例患者，分为 mnLC 组（n=30）、良性肺结节（BLN）组（n=27）和健康对照组（n=30）。采用透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和 Western blot（WB）测量 sEVs 的形态特征和表面标志物。体外分析采用实时定量聚合酶链反应（RT-qPCR）、CCK-8 细胞增殖试验、克隆形成试验、Transwell、干细胞球形成试验和 WB 试验，分别验证 miR-412-3p/TEAD1 信号轴对肺癌细胞生物学功能的影响。进一步通过参与者的血清 sEVs 进行验证。

结果

mnLC 组 sEVs-miR-412-3p 表达水平明显高于 BLN 组和健康对照组（P<0.01）。在肺癌细胞系中，miR-412-3p 可负向调控靶基因 TEAD1。miR-412-3p/TEAD1 信号轴参与促进 EMT 信号通路，并调节肺癌细胞增殖、迁移和干性的恶性生物学功能（P<0.05）。此外，mnLC 组 sEVs 明显促进肺癌细胞增殖、迁移和干性，抑制肺癌细胞 E-cadherin 和 TEAD1 表达，促进 N-cadherin 和 Vimentin 表达（P<0.05）。