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Updated: Aug 9, 2025
A Biomimetic Model for Liver Cancer to Study Tumor-Stroma Interactions in a 3D Environment with Tunable Bio-Physical Properties
Published on: August 7, 2020
Network Analysis for the Discovery of Common Oncogenic Biomarkers in Liver Cancer Experimental Models
Loraine Kay D Cabral1,2, Pablo J Giraudi1, Gianluigi Giannelli3
1Fondazione Italiana Fegato ONLUS, AREA Science Park, Campus Basovizza, 34149 Trieste, Italy.
Abstract:
Hepatocellular carcinoma (HCC) is a malignancy marked by heterogeneity. This study aimed to discover target molecules for potential therapeutic efficacy that may encompass HCC heterogeneity. In silico analysis using published datasets identified 16 proto-oncogenes as potential pharmacological targets. We used an immortalized hepatocyte (IHH) and five HCC cell lines under two subtypes: S1/TGFβ-Wnt-activated (HLE, HLF, and JHH6) and the S2/progenitor subtype (HepG2 and Huh7). Three treatment modalities, 5 µM 5-Azacytidine, 50 µM Sorafenib, and 20 nM PD-L1 gene silencing, were evaluated in vitro. The effect of treatments on the proto-oncogene targets was assessed by gene expression and Western blot analysis. Our results showed that 10/16 targets were upregulated in HCC cells, where cells belonging to the S2/progenitor subtype had more upregulated targets compared to the S1/TGFβ-Wnt-activated subtype (81% vs. 62%, respectively). Among the targets, FGR was consistently down-regulated in the cell lines following the three different treatments. Sorafenib was effective to down-regulate targets in S2/progenitor subtype while PD-L1 silencing was able to decrease targets in all HCC subtypes, suggesting that this treatment strategy may comprise cellular heterogeneity. This study strengthens the relevance of liver cancer cellular heterogeneity in response to cancer therapies.
Insights
This study identified proto-oncogenes as therapeutic targets for hepatocellular carcinoma (HCC), finding PD-L1 gene silencing effective across HCC subtypes, addressing tumor heterogeneity.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Hepatocellular carcinoma (HCC) exhibits significant cellular heterogeneity.
- Understanding this heterogeneity is crucial for developing effective therapeutic strategies.
- Proto-oncogenes represent potential targets for novel HCC treatments.
Purpose of the Study:
- To identify proto-oncogenes as potential therapeutic targets in HCC.
- To investigate the differential expression of these targets across HCC subtypes.
- To evaluate the efficacy of different treatment modalities in modulating target gene expression.
Main Methods:
- In silico analysis of published datasets to identify proto-oncogene targets.
- In vitro experiments using immortalized hepatocytes and HCC cell lines (S1 and S2 subtypes).
- Treatment with 5-Azacytidine, Sorafenib, and PD-L1 gene silencing, followed by gene expression and Western blot analysis.
Main Results:
- Ten out of sixteen identified proto-oncogenes were upregulated in HCC cells.
- The S2/progenitor subtype showed higher target upregulation (81%) compared to the S1/TGFβ-Wnt-activated subtype (62%).
- FGR was consistently downregulated across all treatments; PD-L1 silencing effectively reduced targets in all HCC subtypes.
Conclusions:
- Proto-oncogene dysregulation contributes to HCC heterogeneity.
- PD-L1 gene silencing demonstrates potential for broad efficacy across diverse HCC subtypes.
- Targeting proto-oncogenes and considering cellular heterogeneity are vital for advancing HCC therapies.
