Abstract
Background
Hypoxia contributes to cancer progression through various molecular mechanisms and hepatocellular carcinoma (HCC) is one of the most hypoxic malignancies. Hypoxia-inducible gene domain protein-1a (HIGD1A) is typically induced via epigenetic regulation and promotes tumor cell survival during hypoxia. However, the role of HIGD1A in HCC remains unknown.
Methods
HIGD1A expression was determined in 24 pairs of human HCC samples and para-tumorous tissues. Loss-of-function experiments were conducted both in vivo and in vitro to explore the role of HIGD1A in HCC proliferation and metastasis.
Results
Increased HIGD1A expression was found in HCC tissues and cell lines, which was induced by hypoxia or low-glucose condition. Moreover, HIGD1A knockdown in HCC cells arrested the cell cycle at the G2/M phase and promoted hypoxia-induced cell apoptosis, resulting in great inhibition of cell proliferation, migration, and invasion, as well as tumor xenograft formation. Interestingly, these anti-tumor effects were not observed in normal hepatocyte cell line L02. Further, HIGD1A knockdown suppressed the expression of ornithine decarboxylase 1 (ODC1), a rate-limiting enzyme of polyamine metabolism under c-Myc regulation. HIGD1A was found to bind with the c-Myc promoter region, and its knockdown decreased the levels of polyamine metabolites. Consistently, the inhibitory effect on HCC phenotype by HIGD1A silencing could be reversed by overexpression of c-Myc or supplementation of polyamines.
Conclusions
Our results demonstrated that HIGD1A activated c-Myc–ODC1 nexus to regulate polyamine synthesis and to promote HCC survival and malignant phenotype, implying that HIGD1A might represent a novel therapeutic target for HCC.
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Background
Hepatocellular carcinoma (HCC), a major type of liver cancer, ranks fourth in global incidence and third in mortality among malignant tumors [1]. In the past decade, despite notable advancements in novel drugs and therapies within the field of liver cancer treatment, along with progress in concepts such as early screening, integrated prevention, and collaboration treatment, the overall prognosis for liver cancer remains dismal [2]. The 5-year overall survival rate is only 12.1%, with merely 30% of patients having the opportunity for surgical resection [3]. Previous studies have indicated that tumor recurrence, metastasis, and drug resistance were the major causes of poor prognosis in HCC patients [4]. Therefore, further exploring the molecular mechanisms that participate in HCC development and progression would be useful for identifying novel drug targets.
HCC is among the most hypoxic malignancies, particularly within areas of tumor necrosis [5]. Moreover, treatments for HCC, such as transarterial chemoembolization that restricts blood supply to inhibit tumor growth, or tyrosine kinase inhibitors that reduce angiogenesis by inhibiting various kinase targets, can further aggravate intratumoral hypoxia [6,7,8]. Many studies revealed that within HCC tissues, the expression levels of hypoxia-inducible factors (such as HIF-1α, HIF-2α) or their target genes (GLUT1, LDHA, CA9, SLC7A1) are significantly elevated in comparison to non-tumorous liver tissue. Moreover, this elevated expression was linked to tumor recurrence, metastasis, and reduced patient survival rates [9, 10]. Functional research demonstrated that the signaling cascade of HIF molecules was involved in HCC cell proliferation, angiogenesis, invasion, and metastasis [11, 34]. Certainly, further experimental validation is needed to confirm this hypothesis.
Polyamines were essential for cell proliferation and survival, and both the growth of tumor cells and tumor progression require elevated levels of polyamines [35]. Inhibiting polyamine metabolic processes or reducing polyamine metabolic products can exert anti-tumor effects [36]. ODC1 is a rate-limiting enzyme in the downstream polyamine biosynthetic pathway [37]. In liver cancer, the expression and activity of ODC1 were elevated compared to normal tissues [29]. Difluoromethylornithine is a highly effective and specific inhibitor of ODC1 [38] and its treatment resulted in intracellular polyamine depletion, cell cycle arrest [39], thereby inhibiting tumor cell growth. Considering the significant decrease in the expression of c-Myc and ODC1 following HIGD1A silencing in HCC cells, it suggested that the intracellular polyamine metabolic pathway was inhibited, leading to a reduction in polyamine products. This inhibition consequently suppressed HCC cell proliferation and other phenotypes. Based on the above observation, we speculated that the polyamine metabolic products might serve as the effector molecules through which HIGD1A regulated the phenotypes of HCC cells. Therefore, our results further confirmed the proposed strategy that targets c-Myc and ODC1 in HCC treatment.
Conclusions
Current anti-tumor therapies for HCC remain unsatisfactory, predominantly due to their limited efficacy in blocking the high rates of recurrence and metastasis. Our study has revealed that targeted modulation of HIGD1A yields a notable inhibition to the proliferative, migratory, and invasive capacities of HCC cells. This effect is associated with polyamine levels regulated by the c-Myc–ODC1 interplay. Importantly, our study reveals the discrepancies in HIGD1A expressions and functionalities between HCC cells and their normal counterparts. This discernment helps us to identify the underlying mechanisms driving HCC tumorigenesis. Moreover, it implies a promising target for HCC therapeutic intervention, suggesting that the development of HIGD1A inhibitors could be potentially used for the treatment of liver cancer.
Availability of data and materials
All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the corresponding authors.
Abbreviations
- HCC:
-
Hepatocellular carcinoma
- HIGD1A:
-
Hypoxia-inducible gene domain protein-1a
- ODC1:
-
Ornithine decarboxylase 1
- HIFs:
-
Hypoxia-inducible factors
- CHIP:
-
Chromatin immunoprecipitation
- HPLC:
-
High-Performance Liquid Chromatography
- DNMT1:
-
DNA methyl transferase 1
- CHIP:
-
Chromatin immunoprecipitation
- APH:
-
Aphidicolin
- OCR:
-
Oxygen Consumption Rate
- ECAR:
-
Extracellular Acidification Rate
- EMT:
-
Epithelial-to-mesenchymal transition
- GSEA:
-
Gene set enrichment analysis
- DEGs:
-
Differentially expressed genes
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Acknowledgments
We would like to thank Prof. Chuanjiang Li (Division of Hepatobiliopancreatic Surgery, Department of General Surgery, Nanfang Hospital, Southern Medical University) for providing the clinical samples.
Funding
This study was supported by grants from the National Key Research and Development Program of China (No.2022YFC2303603), the National Natural Science Foundation of China (82172257), and the Health Care Major Project of Guangzhou (202206080001).
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HYL, XYZ, and JLH conceived and designed the study. HXZ, XRL, ZYL, ZML, KYH, YRW, YC, LYL, LYW, and ZLX performed the experiments, data collection, and statistical analysis. HXZ, XRL, and ZYL analyzed the data and wrote the manuscript. XYZ and HYL contributed to data interpretation and critical revision of the manuscript. All authors reviewed and approved the final version of the manuscript.
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This study was approved by the Southern Medical University of Nanfang Hospital (Approval No. 2019-029) and informed consent was obtained from all patients. All animal protocols were approved by the Animal Ethics Committee of the Nanfang Hospital of Southern Medical University (Permit No. IACUC-LAC-20221009-006).
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Zhang, H., Li, X., Liu, Z. et al. Elevated expression of HIGD1A drives hepatocellular carcinoma progression by regulating polyamine metabolism through c-Myc–ODC1 nexus. Cancer Metab 12, 7 (2024). https://doi.org/10.1186/s40170-024-00334-6
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DOI: https://doi.org/10.1186/s40170-024-00334-6