Department of Gastrointestinal and Colorectal Surgery, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China.
Department of Oncology, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China.
Gene. 2023 Jun 20;870:147413. doi: 10.1016/j.gene.2023.147413. Epub 2023 Apr 5.
Colorectal cancer (CRC) is among the most frequent tumors of the digestive tract and the second leading cause of cancer death worldwide. Tumor-associated macrophages (TAMs) are one of the most critical immune cells in the tumor microenvironment, which closely interact with tumor cells to promote tumor incidence and progression. However, the precise mechanism of action between CRC cells and TAMs polarization is still being investigated.
Transmission electronic microscopy (TEM), NanoSight and western blotting were used to characterize exosomes (Exo) isolated from the culture medium of CRC cells. The cellular uptake and internalization of Exo were detected by confocal laser scanning microscopy. M1/ M2 phenotype markers expression were examined by ELISA and flow cytometry. Cell migration, invasion and proliferation were determined by transwell and CCK-8 assay, respectively. A xenograft tumor model was established to explore the role of circVCP in vivo. The target genes of circVCP or miR-9-5p were predicted by StarBase2.0. The target association among miR-9-5p and circVCP or NRP1 was confirmed using the luciferase assay and RNA-pull down assay.
circVCP was highly accumulated in exosomes derived from plasma of CRC patients and CRC cells. Additionally, exosomal circVCP derived from CRC cells promoted cell proliferation, migration and invasion by regulating the miR-9-5p/NRP1 axis, and induced macrophage M2 polarization and inhibited macrophage M1 polarization.
Over-expressed exosomal circVCP promoted the progression of CRC by regulating macrophage M1/M2 polarization through miR-9-5p/NRP1 axis. CircVCP may be a diagnostic biomarker and potential target for CRC therapy.
结直肠癌（CRC）是消化道最常见的肿瘤之一，也是全球癌症死亡的第二大主要原因。肿瘤相关巨噬细胞（TAMs）是肿瘤微环境中最关键的免疫细胞之一，它们与肿瘤细胞密切相互作用，促进肿瘤的发生和发展。然而，CRC 细胞与 TAMs 极化之间的确切作用机制仍在研究中。
使用透射电子显微镜（TEM）、纳米粒子跟踪分析（NanoSight）和蛋白质印迹法来表征从 CRC 细胞培养物中分离的外泌体（Exo）。通过共聚焦激光扫描显微镜检测 Exo 的细胞摄取和内化。通过酶联免疫吸附试验（ELISA）和流式细胞术检测 M1/M2 表型标志物的表达。通过 Transwell 和 CCK-8 测定分别测定细胞迁移、侵袭和增殖。建立异种移植肿瘤模型来体内研究 circVCP 的作用。通过 StarBase2.0 预测 circVCP 或 miR-9-5p 的靶基因。使用荧光素酶测定和 RNA 下拉测定证实了 miR-9-5p 和 circVCP 或 NRP1 之间的靶基因关联。
CRC 患者和 CRC 细胞血浆来源的外泌体中高度积累了 circVCP。此外，来自 CRC 细胞的外泌体 circVCP 通过调节 miR-9-5p/NRP1 轴促进细胞增殖、迁移和侵袭，并诱导巨噬细胞 M2 极化和抑制巨噬细胞 M1 极化。
过表达的外泌体 circVCP 通过 miR-9-5p/NRP1 轴调节巨噬细胞 M1/M2 极化促进 CRC 的进展。CircVCP 可能是 CRC 诊断的生物标志物和潜在的治疗靶点。
