外泌体 LncRNA LINC00659 来源于肿瘤相关成纤维细胞，通过 miR-342-3p/ANXA2 轴促进结直肠癌细胞的进展。

Exosomal LncRNA LINC00659 transferred from cancer-associated fibroblasts promotes colorectal cancer cell progression via miR-342-3p/ANXA2 axis.

机构信息

Departmemt of General Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.

Department of Geriatric Medicine, Xiangya Hospital, Central South University, No. 87, Xiangya Road, Kaifu District, Changsha, Hunan, 410008, People's Republic of China.

出版信息

J Transl Med. 2021 Jan 6;19(1):8. doi: 10.1186/s12967-020-02648-7.

DOI:10.1186/s12967-020-02648-7

PMID:33407563

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7789760/

Abstract

BACKGROUND

Cancer-associated fibroblasts (CAFs) play a pivotal role in regulating tumor progression by transferring exosomes to adjacent cells. Our aim was to clarify the role of LINC00659 encapsulated in CAFs-derived exosomes (CAFs-exo) in colorectal cancer (CRC).

METHODS

CAFs and normal fibroblasts (NFs) were isolated and cultured. CAFs-exo and NFs-derived exosomes (NFs-exo) were characterized by transmission electron microscope and Western blot. The mRNA level of LINC00659 in CAFs-exo and NFs-exo were measured. Then we analyzed cell proliferation by CCK-8 and clone formation assay, cell migration by cell scratch, and cell invasion by Transwell. Epithelial mesenchymal transformation (EMT) related markers E-cadherin, N-cadherin, Vimentin and Snail-1 expressions were assessed by Western blot. The binding of LINC00659 and miR-342-3p, miR-342-3p and ANXA2 were analyzed by dual-luciferase reporter gene assay.

RESULTS

CAFs and NFs showed a spindle-like morphology. CAFs-exo promoted CRC cell proliferation, migration, invasion and EMT progression. The expression of LINC00659 in CAF-derived exosomes was significantly increased, and fibroblasts could transfer exosomal LINC00659 to CRC cells. We further revealed that transfection of miR-342-3p mimic or sh-ANXA2 could obviously reverse the promotion effect of exosomal LINC00659 on CRC progression. Functional studies reveal that LINC00659 is transferred from CAFs to the cancer cells via exosomes, where it promotes CRC cell proliferation, invasion, migration and EMT progression in vitro. Mechanistically, LINC00659 interacts directly with miR-342-3p to increase ANXA2 expression in CRC cells.

CONCLUSION

Collected evidence supported that CAFs-derived exosomal LINC00659 promotes CRC cell proliferation, invasion and migration via miR-342-3p/ANXA2axis.

摘要

背景

癌症相关成纤维细胞（CAFs）通过将外泌体转移到邻近细胞，在调节肿瘤进展方面发挥关键作用。我们的目的是阐明包裹在 CAFs 衍生的外泌体（CAFs-exo）中的 LINC00659 在结直肠癌（CRC）中的作用。

方法

分离并培养 CAFs 和正常成纤维细胞（NFs）。通过透射电子显微镜和 Western blot 对 CAFs-exo 和 NFs 衍生的外泌体（NFs-exo）进行表征。测量 CAFs-exo 和 NFs-exo 中 LINC00659 的 mRNA 水平。然后通过 CCK-8 和克隆形成实验分析细胞增殖，通过细胞划痕分析细胞迁移，通过 Transwell 分析细胞侵袭。通过 Western blot 评估上皮间质转化（EMT）相关标志物 E-钙粘蛋白、N-钙粘蛋白、波形蛋白和 Snail-1 的表达。通过双荧光素酶报告基因检测分析 LINC00659 与 miR-342-3p、miR-342-3p 与 ANXA2 的结合。

结果

CAFs 和 NFs 呈纺锤形。CAFs-exo 促进 CRC 细胞增殖、迁移、侵袭和 EMT 进展。CAF 衍生的外泌体中 LINC00659 的表达显著增加，成纤维细胞可以将外泌体 LINC00659 转移到 CRC 细胞。我们进一步表明，转染 miR-342-3p 模拟物或 sh-ANXA2 可以明显逆转外泌体 LINC00659 对 CRC 进展的促进作用。功能研究表明，LINC00659 通过外泌体从 CAFs 转移到癌细胞，在体外促进 CRC 细胞的增殖、侵袭、迁移和 EMT 进展。机制上，LINC00659 与 miR-342-3p 直接相互作用，增加 CRC 细胞中 ANXA2 的表达。