Theranostics 2019; 9(12):3639-3652. doi:10.7150/thno.31693

Research Paper

Integration of metabolomics and expression of enolase-phosphatase 1 links to hepatocellular carcinoma progression

Hao Zhuang1,2*, Zhaoyan Qiang3*, Xiaowen Shao2*, Huan Wang4, Yamei Dang2, Zun Wang2, Fei Wu2, Wen Wei4✉, Yongmei Li2✉

1. Department of Hepatic Biliary Pancreatic Surgery, Cancer Hospital Affiliated to Zhengzhou University, Zhengzhou, Henan Province 450000, China
2. Department of Pathogen Biology, School of Basic Medical Sciences, Key Lab of Immune Microenvironment and Disease (Ministry of Education) Tianjin Medical University, Tianjin 300070, China
3. Department of Pharmacology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China
4. School of Life Sciences, Chongqing University, Chongqing, 400044, China
*These authors contribute equally to this manuscript.

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license ( See for full terms and conditions.
Zhuang H, Qiang Z, Shao X, Wang H, Dang Y, Wang Z, Wu F, Wei W, Li Y. Integration of metabolomics and expression of enolase-phosphatase 1 links to hepatocellular carcinoma progression. Theranostics 2019; 9(12):3639-3652. doi:10.7150/thno.31693. Available from

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Reprogramming of cellular metabolism is one of the hallmarks for cancer, in which tumor cells rewire their metabolic fluxes to generate sufficient energy and biosynthetic intermediates. Therefore, elucidating the correlation between cellular metabolism and hepatocellular carcinoma (HCC) progression may provide insights into novel approaches to cancer therapy.

Methods: We assembled an integrated pathway-level metabolic profiling by mining metabolomic, transcriptomic and proteomic data of three HCC cell lines with increasing metastatic potentials. Immunohistochemical staining was performed in a tissue microarray from 185 HCC clinical specimens. Kaplan-Meier survival and Cox regression analyses were applied to test the association between gene expression and survival outcome. In vitro assays were conducted to investigate the functional role of enolase-phosphatase 1 (ENOPH1) in HCC malignant behaviors. Reversed genetics analysis was performed to determine the function of ENOPH1 in HCC metastasis. An intrahepatic mouse model further confirmed the role of ENOPH1 in metastasis.

Results: We have determined that HCC cell metastasis is associated with alterations in metabolite levels and expressions of metabolic enzymes in the cysteine/methionine metabolism pathway, and show that one of metabolic enzymes, enolase-phosphatase 1 (ENOPH1), is persistently upregulated with an increase in metastatic potential. The upregulation of ENOPH1 expression was observed as an independent prognostic factor for HCC patients. ENOPH1 overexpression promoted cell migration and invasion, whereas ENOPH1 downregulation inhibited cell migration and invasion. Furthermore, an enhanced phosphorylation of AKT with ENOPH1 upregulation was observed. ENOPH1-mediated malignant capacity in HCC cells can be rescued by an AKT inhibitor.

Conclusion: Taken together, our findings illustrate that ENOPH1 promotes HCC progression and could serve as a novel biomarker and therapeutic target for HCC.

Keywords: hepatocellular carcinoma, enolase-phosphatase 1, AKT, omics technologies