CAFs 来源的小细胞外囊泡 circN4BP2L2 通过 miR-664b-3p/HMGB3 通路促进结直肠癌细胞的增殖和转移。

CAFs-derived small extracellular vesicles circN4BP2L2 promotes proliferation and metastasis of colorectal cancer via miR-664b-3p/HMGB3 pathway.

机构信息

Department of Pathology, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.

Department of Gynecology, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.

出版信息

Cancer Biol Ther. 2022 Dec 31;23(1):404-416. doi: 10.1080/15384047.2022.2072164.

Our previous research has demonstrated that colorectal cancer (CRC) progression was promoted by circN4BP2L2. This study aimed to further explore the mechanism of circN4BP2L2 in the development of CRC from the perspective of small extracellular vesicles (sEVs). Cancer-associated fibroblasts cell (CAFs) and normal fibroblasts cell (NFs) were isolated from CRC tissues and adjacent tissues, respectively. The ultra-centrifugation was used for extraction of their related sEVs. Cell proliferation and apoptosis were analyzed using CCK-8 and flow cytometry, respectively. Transwell assay was conducted to measure cell migration. The tube formation ability was assessed by tube formation assay. The target relationships between circN4BP2L2 and miR-664b-3p, and miR-664b-3p and HMGB3 were validated by dual-luciferase reporter detection. The effect of CAFs-derived sEV (CAFs-sEVs) circN4BP2L2 on CRC was further studied in nude mice. CAFs-exo promoted cell proliferation, migration, tube formation ability, and inhibited apoptosis of CRC cells. CAFs-sEV circN4BP2L2 knockdown reversed the above results. CircN4BP2L2 directly targeted miR-664b-3p, and HMGB3 was targeted by miR-664b-3p. Moreover, subcutaneous tumorigenesis and liver metastasis of nude mice with CRC were repressed by CAFs-sEV circN4BP2L2 knockdown. CAFs-sEV circN4BP2L2 knockdown restrained CRC cell proliferation and migration by regulating miR-664b-3p/HMGB3 pathway.

我们之前的研究表明，结直肠癌（CRC）的进展是由 circN4BP2L2 促进的。本研究旨在从小细胞外囊泡（sEVs）的角度进一步探讨 circN4BP2L2 在 CRC 发展中的机制。分别从 CRC 组织和相邻组织中分离出癌相关成纤维细胞（CAFs）和正常成纤维细胞（NFs），通过超速离心提取其相关 sEV。使用 CCK-8 和流式细胞术分别分析细胞增殖和凋亡。Transwell 测定用于测定细胞迁移。通过管形成测定评估管形成能力。通过双荧光素酶报告检测验证 circN4BP2L2 与 miR-664b-3p 以及 miR-664b-3p 与 HMGB3 之间的靶标关系。进一步在裸鼠中研究了 CAFs 衍生的 sEV（CAFs-sEVs）circN4BP2L2 对 CRC 的影响。CAFs-exo 促进 CRC 细胞增殖、迁移、管形成能力，并抑制细胞凋亡。CAFs-sEV circN4BP2L2 敲低逆转了上述结果。CircN4BP2L2 直接靶向 miR-664b-3p，而 miR-664b-3p 靶向 HMGB3。此外，CAFs-sEV circN4BP2L2 敲低抑制了裸鼠 CRC 皮下肿瘤生成和肝转移。CAFs-sEV circN4BP2L2 敲低通过调节 miR-664b-3p/HMGB3 通路抑制 CRC 细胞增殖和迁移。