肥胖小鼠脂肪细胞来源的细胞外囊泡运输 miR-99a-5p 以减轻非小细胞肺癌细胞的增殖和迁移。

Obese Mouse Fat Cell-derived Extracellular Vesicles Transport miR-99a-5p to Mitigate the Proliferation and Migration of Non-small Cell Lung Cancer Cells.

机构信息

Department of General Practice, Yantai Yuhuangding Hospital Affiliated to Qingdao University, Yantai, 264000, China.

Department of Pulmonary and Critical Care Medicine, Yantai Yuhuangding Hospital Affiliated to Qingdao University, Yantai, 264000, China.

出版信息

Comb Chem High Throughput Screen. 2024;27(2):214-226. doi: 10.2174/1386207326666230316103604.

DOI:10.2174/1386207326666230316103604

PMID:36927435

Abstract

OBJECTIVE

Fat cells-derived extracellular vesicles (FC-EVs) play a role in regulating the tumor microenvironment in cancers by transporting RNAs. MicroRNAs (miRNAs) are vital regulators of cancer development. This study was conducted to explore the role of FC-EVs in the proliferation and migration of non-small cell lung cancer (NSCLC) cells, providing targets for NSCLC treatment.

METHODS

The obese mouse model was established high-fat diet (HFD), followed by separation and characterization of FC-EVs (HFD-EVs). The levels of miR-99a-5p, precursor-miR-99a-5p, and heparan sulfate-glucosamine 3-sulfotransferase 3B1 (HS3ST3B1) were measured by RT-qPCR or Western blot assay. Cell proliferation and migration were evaluated by 3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide and wound healing assays. The expression of Cy3-labeled miR-99a-5p in A549 cells (one NSCLC cell line) was observed confocal microscopy. The binding of miR-99a-5p to HS3ST3B1 was analyzed by the dual luciferase assay. Rescue experiments were performed to confirm the role of HS3ST3B1 in NSCLC cells.

RESULTS

miR-99a-5p was upregulated in adipose tissues, FCs, and HFD-EVs. HFD-EVs mitigated the proliferation and migration of NSCLC cells. HFD-EVs transported miR-99a-5p into A549 cells, which upregulated miR-99a-5p expression and inhibited HS3ST3B1 expression in A549 cells. HS3ST3B1 overexpression reversed the inhibition of HFD-EVs on the proliferation and migration of NSCLC cells.

CONCLUSION

HFD-EVs transported miR-99a-5p into NSCLC cells and inhibited HS3ST3B1, thereby inhibiting proliferation and migration of NSCLC cells.

摘要

目的

脂肪细胞衍生的细胞外囊泡（FC-EVs）通过转运 RNA 在癌症中发挥调节肿瘤微环境的作用。微小 RNA（miRNA）是癌症发展的重要调节因子。本研究旨在探讨 FC-EVs 在非小细胞肺癌（NSCLC）细胞增殖和迁移中的作用，为 NSCLC 的治疗提供靶点。

方法

建立肥胖小鼠模型（高脂肪饮食，HFD），分离和表征 FC-EVs（HFD-EVs）。通过 RT-qPCR 或 Western blot 测定 miR-99a-5p、前体 miR-99a-5p 和硫酸乙酰肝素葡糖胺 3-O-磺基转移酶 3B1（HS3ST3B1）的水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐和划痕愈合实验评估细胞增殖和迁移。通过共聚焦显微镜观察 Cy3 标记的 miR-99a-5p 在 A549 细胞（一种 NSCLC 细胞系）中的表达。通过双荧光素酶报告基因实验分析 miR-99a-5p 与 HS3ST3B1 的结合。进行挽救实验以确认 HS3ST3B1 在 NSCLC 细胞中的作用。

结果

miR-99a-5p 在脂肪组织、FCs 和 HFD-EVs 中上调。HFD-EVs 减轻了 NSCLC 细胞的增殖和迁移。HFD-EVs 将 miR-99a-5p 转运到 A549 细胞中，上调 miR-99a-5p 的表达并抑制 A549 细胞中的 HS3ST3B1 表达。HS3ST3B1 过表达逆转了 HFD-EVs 对 NSCLC 细胞增殖和迁移的抑制作用。