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Updated: Aug 10, 2026
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
Microarray profiling shows distinct differences between primary tumors and commonly used preclinical models in
Weining Wang1, N Gopalakrishna Iyer2,3, Hsien Ts'ung Tay4
1Cellular and Molecular Research, National Cancer Centre, Singapore, 169610, Singapore. wang.weining@u.duke.nus.edu.
Background:
Despite advances in therapeutics, outcomes for hepatocellular carcinoma (HCC) remain poor and there is an urgent need for efficacious systemic therapy. Unfortunately, drugs that are successful in preclinical studies often fail in the clinical setting, and we hypothesize that this is due to functional differences between primary tumors and commonly used preclinical models. In this study, we attempt to answer this question by comparing tumor morphology and gene expression profiles between primary tumors, xenografts and HCC cell lines.
Methods:
Hep G2 cell lines and tumor cells from patient tumor explants were subcutaneously (ectopically) injected into the flank and orthotopically into liver parenchyma of Mus Musculus SCID mice. The mice were euthanized after two weeks. RNA was extracted from the tumors, and gene expression profiling was performed using the Gene Chip Human Genome U133 Plus 2.0. Principal component analyses (PCA) and construction of dendrograms were conducted using Partek genomics suite.
Results:
PCA showed that the commonly used HepG2 cell line model and its xenograft counterparts were vastly different from all fresh primary tumors. Expression profiles of primary tumors were also significantly divergent from their counterpart patient-derived xenograft (PDX) models, regardless of the site of implantation. Xenografts from the same primary tumors were more likely to cluster together regardless of site of implantation, although heat maps showed distinct differences in gene expression profiles between orthotopic and ectopic models.
Conclusions:
The data presented here challenges the utility of routinely used preclinical models. Models using HepG2 were vastly different from primary tumors and PDXs, suggesting that this is not clinically representative. Surprisingly, site of implantation (orthotopic versus ectopic) resulted in limited impact on gene expression profiles, and in both scenarios xenografts differed significantly from the original primary tumors, challenging the long-held notion that orthotopic PDX model is the gold standard preclinical model for HCC.
Insights
Hepatocellular carcinoma (HCC) preclinical models, including HepG2 and patient-derived xenografts (PDXs), do not accurately represent primary tumors. Gene expression profiles reveal significant differences, challenging their clinical utility for developing new HCC therapies.
Area of Science:
- Oncology
- Translational Research
- Genomics
Background:
- Hepatocellular carcinoma (HCC) outcomes remain poor despite therapeutic advances.
- Effective systemic therapies are urgently needed for HCC.
- Preclinical models often fail to predict clinical efficacy due to functional differences from primary tumors.
Purpose of the Study:
- To compare gene expression profiles and tumor morphology between primary HCC tumors, xenografts, and cell lines.
- To assess the clinical relevance of commonly used HCC preclinical models.
Main Methods:
- HepG2 cell lines and patient tumor explants were implanted subcutaneously and orthotopically in mice.
- Gene expression profiling was performed on extracted RNA.
- Principal component analyses (PCA) and dendrograms were used for data analysis.
Main Results:
- HepG2 models and their xenografts significantly differed from primary HCC tumors.
- Patient-derived xenografts (PDXs) showed divergent expression profiles from their original primary tumors.
- Gene expression profiles of xenografts showed limited impact from implantation site (orthotopic vs. ectopic).
Conclusions:
- Commonly used HCC preclinical models, including HepG2 and PDXs, lack clinical representativeness.
- The site of implantation had minimal effect on xenograft gene expression.
- These findings challenge the utility of current preclinical models and the notion of orthotopic PDXs as the gold standard for HCC research.
