Chronic stress–induced ANPEP drives liver cancer progression by increasing glutathione synthesis and inhibiting ferroptosis
Yongkang Wu, Yankun Zhang, Xiaojia Shi, Mengting Wu, Min Sun, Ying Feng, Wenmeng Ma, Xiule Jiang, Dingqi Fei, Mingjian Zhao, Zhuanchang Wu, Chunyang Li, Xiaohong Liang, Lifen Gao, Chunhong Ma, Xuetian Yue
Yongkang Wu, Yankun Zhang, Xiaojia Shi, Mengting Wu, Min Sun, Ying Feng, Wenmeng Ma, Xiule Jiang, Dingqi Fei, Mingjian Zhao, Zhuanchang Wu, Chunyang Li, Xiaohong Liang, Lifen Gao, Chunhong Ma, Xuetian Yue
View: Text | PDF
Research Article Cell biology Gastroenterology Hepatology

Chronic stress–induced ANPEP drives liver cancer progression by increasing glutathione synthesis and inhibiting ferroptosis

Abstract

Emerging evidence demonstrates that chronic stress alters immunological, neurochemical, and endocrinological functions, thereby promoting tumor progression. However, the underlying metabolic mechanism of chronic stress in tumor progression is still elusive. Using multiomics analysis, we found that aminopeptidase N (ANPEP) was upregulated in tumors with chronic restraint, associating with the reprogramming of amino acid metabolism. Functional assays revealed that ANPEP promoted liver cancer growth and metastasis. Knockdown of ANPEP blocked chronic stress–induced liver cancer progression. Chronic stress–induced glucocorticoids promoted nuclear receptor subfamily 3 group C member 1 nuclear translocation to activate ANPEP transcription by directly binding to its promoter. Furthermore, ANPEP promotes glutathione synthesis, subsequently inhibiting ROS-induced ferroptosis. Mechanistically, ANPEP interacted with solute carrier family 3 member 2 (SLC3A2) to block membrane associated ring-CH-type finger 8–mediated lysosome-dependent degradation of SLC3A2, promoting intracellular l-cystine transport, thereby increasing glutathione synthesis. The combination of ANPEP silencing and sorafenib treatment showed a synergistic effect in inhibiting liver cancer progression. Finally, clinical data and mouse models demonstrated that chronic stress drove liver tumor progression via ANPEP-regulated SLC3A2. These findings reveal unanticipated communication between chronic stress and metabolic reprogramming during liver cancer progression, providing potential therapeutic implications for liver cancer.

Authors

Yongkang Wu, Yankun Zhang, Xiaojia Shi, Mengting Wu, Min Sun, Ying Feng, Wenmeng Ma, Xiule Jiang, Dingqi Fei, Mingjian Zhao, Zhuanchang Wu, Chunyang Li, Xiaohong Liang, Lifen Gao, Chunhong Ma, Xuetian Yue

×

Figure 5

ANPEP enhances GSH levels to reduce sorafenib sensitivity of liver cancer.

Options: View larger image (or click on image) Download as PowerPoint
ANPEP enhances GSH levels to reduce sorafenib sensitivity of liver cance...
(A) Targeted amino acid metabolomic profiling of Huh7 cells with or without ANPEP overexpression, followed by KEGG pathway enrichment analysis of differential metabolites. (B and C) GSH levels and GSH/GSSG ratios in liver cancer cells with ANPEP manipulation (B, n = 5; C, n = 3). (D and E) Lipid peroxide levels were labeled by C11-BODIPY in Huh7 and HepG2 cells with ANPEP manipulation (D: Huh7, n = 4; HepG2, n = 5. E: Huh7, n = 4; HepG2, n = 6). (F and G) IC50 of ANPEP-manipulated Huh7 and HepG2 cells with 24 hours of sorafenib treatment (0, 1, 2, 4, 8, 16, 32, 64 μM) (n = 3). (H and I) Lipid peroxides in liver cancer cells treated with sorafenib alone or sorafenib plus ANPEP manipulation, quantified by C11-BODIPY intensity (n = 4). (J) Cell viability of Huh7 and HepG2 cells stably expressing shNC or shANPEP after 24 hours of treatment with sorafenib (10 μM) and/or ferrostatin-1 (2 μM) (n = 4). (K and L) Xenograft tumor model established by subcutaneous injection of ANPEP-manipulated Huh7 cells into nude mice, followed by treatment with sorafenib or vehicle. Tumor growth (K) and MDA and 4-HNE levels (L) were assessed (K, n = 8; L, n = 5). Data are presented as mean ± SD. Statistical significance was determined by 2-way ANOVA with Šidák’s multiple comparisons test (K), 1-way ANOVA with Tukey’s multiple comparisons test (C, E, H–J, and L), and 2-tailed unpaired Student’s t test (B, D, F, and G). **P < 0.01; ***P < 0.001.