Lin Chao, Jiang Tao, E Changyong, Wang Lun, Chen Tong, Wang Xia, Xiang Yien
Hepatobiliary and Pancreatic Surgery, China-Japan Union Hospital of Jilin University, Changchun, China.
Gastrointestinal Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, China.
Cancer Cell Int. 2024 May 31;24(1):193. doi: 10.1186/s12935-024-03380-8.
LncRNA colorectal neoplasia differentially expressed (CRNDE) was found to be an important regulator in many cancers. This project focuses on the function of CRNDE on macrophage metabolic reprogramming and Hepatocellular carcinoma (HCC).
qRT-PCR and Immunofluorescence were used to analyze Arg-1, IL-10, CD163, CCL-18, CD206, and CRNDE expression in HCC tissues and macrophages. Western Blotting was used to analyze ERK and p-ERK expression. Edu assay, transwell assay and xenograft experiments were carried out to study cell viability, migrated and invasive capability. Immunohistochemical staining was used to evaluate Ki67 expression. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed for macrophages metabolites analysis.
Arg-1, IL-10, CD163, CD206, and CRNDE were significantly up-regulated in HCC tissues, M2 macrophage and M0 macrophage with CRNDE overexpressed (OV-CRNDE-M0), which downregulated in M0 macrophage with CRNDE knockdown (sh-CRNDE-M0). The conditioned medium (CM) of M2 cells and OV-CRNDE-M0 cells promoted cell viability, invasion, and migration of HCC cells, the effect was reversed by sh-CRNDE-M0 cells CM. OV-CRNDE-M0 cells promoted tumor growth, Ki67 and CD206 expression in xenograft model. 61 metabolites were detected, of which 18 metabolites changed significantly in OV-CRNDE-M0 group compared to M0 group, with 9 upregulated and 9 downregulated. KEGG analysis showed the enrichment pathways were biosynthesis, glyoxylate and dicarboxylate metabolism. SMPDB analysis showed the enrichment pathways were hypoacetylaspartia, canavan disease, and aspartate metabolism.
CRNDE regulated the metabolic reprogramming of M2 macrophage via ERK pathway, which thereby contributed to HCC proliferation, migration, and invasion.
长链非编码RNA结直肠癌差异表达基因（CRNDE）被发现是多种癌症中的重要调节因子。本项目聚焦于CRNDE在巨噬细胞代谢重编程和肝细胞癌（HCC）中的作用。
采用qRT-PCR和免疫荧光法分析HCC组织及巨噬细胞中精氨酸酶-1（Arg-1）、白细胞介素-10（IL-10）、CD163、CC趋化因子配体18（CCL-18）、CD206和CRNDE的表达。采用蛋白质免疫印迹法分析细胞外信号调节激酶（ERK）和磷酸化细胞外信号调节激酶（p-ERK）的表达。进行EdU检测、Transwell检测和异种移植实验以研究细胞活力、迁移和侵袭能力。采用免疫组织化学染色评估Ki67的表达。利用液相色谱-串联质谱（LC-MS/MS）对巨噬细胞代谢物进行分析。
在HCC组织、M2巨噬细胞和CRNDE过表达的M0巨噬细胞（OV-CRNDE-M0）中，Arg-1、IL-10、CD163、CD206和CRNDE显著上调，而在CRNDE敲低的M0巨噬细胞（sh-CRNDE-M0）中下调。M2细胞和OV-CRNDE-M0细胞的条件培养基（CM）促进了HCC细胞的活力、侵袭和迁移，sh-CRNDE-M0细胞的CM可逆转这种作用。在异种移植模型中，OV-CRNDE-M0细胞促进了肿瘤生长、Ki67和CD206的表达。检测到61种代谢物，其中与M0组相比，OV-CRNDE-M0组中有18种代谢物发生显著变化，9种上调，9种下调。京都基因与基因组百科全书（KEGG）分析显示富集途径为生物合成、乙醛酸和二羧酸代谢。小分子代谢途径数据库（SMPDB）分析显示富集途径为低乙酰天冬氨酸、刀豆氨酸病和天冬氨酸代谢。
CRNDE通过ERK途径调节M2巨噬细胞的代谢重编程，从而促进HCC的增殖、迁移和侵袭。
