酰基辅酶 A 通过 lncBCL2L11-THOC5-JNK 轴促进胆囊癌转移。

Acylcarnitines promote gallbladder cancer metastasis through lncBCL2L11-THOC5-JNK axis.

机构信息

Department of Biliary-Pancreatic Surgery, Renji Hospital Affliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China.

Department of General Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200092, China.

出版信息

J Transl Med. 2024 Mar 22;22(1):299. doi: 10.1186/s12967-024-05091-0.

DOI:10.1186/s12967-024-05091-0

PMID:38519939

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10958842/

Abstract

BACKGROUND

The progression of gallbladder cancer (GBC) is accompanied by abnormal fatty acid β-oxidation (FAO) metabolism. Different types of lipids perform various biological functions. This study aimed to determine the role of acyl carnitines in the molecular mechanisms of GBC progression.

METHODS

Distribution of lipids in GBC was described by LC-MS-based lipidomics. Cellular localization, expression level and full-length of lncBCL2L11 were detected using fluorescence in situ hybridization (FISH) assays, subcellular fractionation assay and 5' and 3' rapid amplification of the cDNA ends (RACE), respectively. In vitro and in vivo experiments were used to verify the biological function of lncBCL2L11 in GBC cells. Methylated RNA Immunoprecipitation (MeRIP) was performed to detect the methylation levels of lncBCL2L11. RNA pull-down assay and RNA immunoprecipitation (RIP) assay were used to identify lncBCL2L11 interacting proteins. Co-Immunoprecipitation (Co-IP) and Western blot assay were performed to validate the regulatory mechanism of lncBCL2L11 and THO complex.

RESULTS

Acylcarnitines were significantly up-regulated in GBC tissues. High serum triglycerides correlated to decreased survival in GBC patients and promoted tumor migration. LncBCL2L11 was identified in the joint analysis of highly metastatic cells and RNA sequencing data. LncBCl2L11 prevented the binding of THOC6 and THOC5 and causes the degradation of THOC5, thus promoting the accumulation of acylcarnitines in GBC cells, leading to the malignant progression of cancer cells. In addition, highly expressed acylcarnitines stabilized the expression of lncBCL2L11 through N-methyladenosine methylation (m6A), forming a positive feedback regulation in tumor dissemination.

CONCLUSIONS

LncBCL2L11 is involved in gallbladder cancer metastasis through FAO metabolism. High lipid intake is associated with poor prognosis of GBC. Therefore, targeting lncBCL2L11 and its pathway-related proteins or reducing lipid intake may be significant for the treatment of GBC patients.

摘要

背景

胆囊癌（GBC）的进展伴随着异常脂肪酸β-氧化（FAO）代谢。不同类型的脂质发挥着不同的生物学功能。本研究旨在确定酰基辅酶 A 在 GBC 进展分子机制中的作用。

方法

通过基于 LC-MS 的脂质组学描述 GBC 中的脂质分布。使用荧光原位杂交（FISH）检测、亚细胞分级测定和 5' 和 3' 快速扩增的 cDNA 末端（RACE）分别检测 lncBCL2L11 的细胞定位、表达水平和全长。在体内和体外实验中验证 lncBCL2L11 在 GBC 细胞中的生物学功能。进行甲基化 RNA 免疫沉淀（MeRIP）以检测 lncBCL2L11 的甲基化水平。使用 RNA 下拉测定和 RNA 免疫沉淀（RIP）测定鉴定 lncBCL2L11 相互作用蛋白。共免疫沉淀（Co-IP）和 Western blot 测定用于验证 lncBCL2L11 和 THO 复合物的调控机制。

结果

酰基辅酶 A 在 GBC 组织中明显上调。高血清甘油三酯与 GBC 患者的生存率降低相关，并促进肿瘤迁移。在高转移性细胞的联合分析和 RNA 测序数据中鉴定出 lncBCL2L11。LncBCl2L11 阻止 THOC6 和 THOC5 的结合并导致 THOC5 的降解，从而促进 GBC 细胞中酰基辅酶 A 的积累，导致癌细胞的恶性进展。此外，高表达的酰基辅酶 A 通过 N-甲基腺苷甲基化（m6A）稳定 lncBCL2L11 的表达，在肿瘤扩散中形成正反馈调节。