Targeting TROY-mediated P85a/AKT/TBX3 signaling attenuates tumor stemness and elevates treatment response in hepatocellular carcinoma

• Published in: Journal of experimental & clinical cancer research 2022-05, Vol.41 (1), p.182-182, Article 182
• Main Authors: Liu, Beilei, Fang, Xiaona, Kwong, Dora Lai-Wan, Zhang, Yu, Verhoeft, Krista, Gong, Lanqi, Zhang, Baifeng, Chen, Jie, Yu, Qianqian, Luo, Jie, Tang, Ying, Huang, Tuxiong, Ling, Fei, Fu, Li, Yan, Qian, Guan, Xin-Yuan
• Format: Article
• Language: English
• Subjects: Analysis; Apoptosis; Cancer; Cancer-associated fibroblast; Carcinoma, Hepatocellular - drug therapy; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - pathology; Cell growth; Cell Line, Tumor; Chemotherapy; Class Ia Phosphatidylinositol 3-Kinase - metabolism; Cloning; Cytokeratin; Drug resistance; Drug therapy; Fibroblasts; Gene expression; Health aspects; Hepatoma; Humans; K63 polyubiquitin modification; Liver; Liver cancer; Liver Neoplasms - drug therapy; Liver Neoplasms - genetics; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Metastasis; Molecular Targeted Therapy; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Oncofetal properties; Pluripotency signaling pathway; Protein Kinase Inhibitors - pharmacology; Proteins; Proto-Oncogene Proteins c-akt - metabolism; Receptors, Tumor Necrosis Factor - metabolism; Signal Transduction - drug effects; Stem cells; T-Box Domain Proteins - metabolism; Transforming growth factors; Wortmannin - pharmacology
• ISSN: 1756-9966; 0392-9078; 1756-9966
• DOI: 10.1186/s13046-022-02401-6

Previous in vitro hepatocyte differentiation model showed that TROY was specifically expressed in liver progenitor cells and a small proportion of hepatocellular carcinoma cells, suggesting that TROY may participate in hepatocellular carcinoma (HCC) stemness regulation. Here, we aim to investigate the role and mechanism of TROY in HCC pathogenesis. Bioinformatics analysis of the TCGA dataset has been used to identify the function and mechanism of TROY. Spheroid, apoptosis, and ALDH assay were performed to evaluate the stemness functions. Validation of the downstream pathway was based on Western blot, co-immunoprecipitation, and double immunofluorescence. HCC tissue microarray study found that a high frequency of TROY-positive cells was detected in 53/130 (40.8%) of HCC cases, which was significantly associated with poor prognosis and tumor metastasis. Functional studies revealed that TROY could promote self-renewal, drug resistance, tumorigenicity, and metastasis of HCC cells. Mechanism study found that TROY could interact with PI3K subunit p85α, inducing its polyubiquitylation and degradation. The degradation of p85α subsequently activate PI3K/AKT/TBX3 signaling and upregulated pluripotent genes expression including SOX2, NANOG, and OCT4, and promoted EMT in HCC cells. Interestingly, immune cell infiltration analysis found that upregulation of TROY in HCC tissues was induced by TGF-β1 secreted from CAFs. PI3K inhibitor wortmannin could effectively impair tumor stemness to sorafenib. We demonstrated that TROY is an HCC CSC marker and plays an important role in HCC stemness regulation. Targeting TROY-positive CSCs with PI3K inhibitor wortmannin combined with chemo- or targeted drugs might be a novel therapeutic strategy for HCC patients.