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Exploratory Multi-Level Analysis of the HIF Axis in Clear-Cell Renal Cell Carcinoma and Evaluation of GN44028 as an
Piotr M Wierzbicki1, Agnieszka Rybarczyk1, Mateusz Czajkowski2
1Department of Histology, Faculty of Medicine, Medical University of Gdańsk, 80-210 Gdańsk, Poland.
Abstract:
Clear-cell renal cell carcinoma (ccRCC) is characterised by constitutive activation of hypoxia-inducible factors (HIFs) following VHL loss, which contributes to tumour progression and therapeutic resistance. Given the limitations of VEGFR-targeted therapies, we investigated the biological and potential therapeutic relevance of the HIF axis in ccRCC. Nuclear and cytoplasmic HIF1A and EPAS1/HIF2A expression were assessed by immunohistochemistry in tumours from 40 patients and correlated with clinicopathological parameters and cancer-specific survival. The functional effects of HIF pathway inhibitors (GN44028, KC7F2, and FM19G11) and sunitinib were analysed in VHL-mutant 786-O and VHL-wild-type Caki-1 cell lines using SRB viability assay, cell cycle analysis, wound closure assay, and RT-qPCR of HIF-related genes, with comparison to non-malignant HK-2 cells. TCGA-ccRCC data from advanced-stage patients (III-IV, n = 185) were analysed as a complementary transcriptomic context. Nuclear, but not cytoplasmic, HIF1A and EPAS1/HIF2A expression was associated with advanced stage and shorter survival in univariable analyses. GN44028 showed the most pronounced antiproliferative effect under tested conditions and was associated with broad suppression of HIF-related transcription, whereas sunitinib was associated with increased expression of selected HIF-related genes. GN44028 did not demonstrate clear selectivity over non-malignant HK-2 cells. Overall, nuclear HIF activation is associated with aggressive ccRCC biology, and broader HIF pathway modulation warrants further experimental investigation; however, the clinical findings remain exploratory, and therapeutic selectivity and translational relevance are not yet established.
Insights
Nuclear HIF activation in clear-cell renal cell carcinoma (ccRCC) correlates with aggressive disease and poor survival. While HIF inhibitors show potential, further research is needed to establish therapeutic selectivity and clinical relevance in ccRCC treatment.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Clear-cell renal cell carcinoma (ccRCC) exhibits constitutive activation of hypoxia-inducible factors (HIFs) due to VHL loss, driving tumor progression and treatment resistance.
- Current VEGFR-targeted therapies for ccRCC have limitations, necessitating exploration of alternative therapeutic strategies targeting the HIF pathway.
Purpose of the Study:
- To investigate the biological and therapeutic relevance of the HIF axis in ccRCC.
- To assess the correlation between nuclear and cytoplasmic HIF1A and EPAS1/HIF2A expression and clinicopathological parameters, including survival.
- To evaluate the functional effects of HIF pathway inhibitors and sunitinib in ccRCC cell lines.
Main Methods:
- Immunohistochemistry was used to assess HIF1A and EPAS1/HIF2A expression in patient tumors.
- In vitro studies utilized ccRCC cell lines (VHL-mutant and VHL-wild-type) and non-malignant cells to analyze the effects of HIF inhibitors (GN44028, KC7F2, FM19G11) and sunitinib.
- Assays included SRB viability, cell cycle analysis, wound closure, and RT-qPCR of HIF-related genes.
- TCGA-ccRCC data from advanced-stage patients were analyzed for transcriptomic context.
Main Results:
- Nuclear HIF1A and EPAS1/HIF2A expression, but not cytoplasmic, was associated with advanced stage and shorter cancer-specific survival.
- GN44028 demonstrated the most significant antiproliferative effect and suppressed HIF-related transcription broadly.
- Sunitinib treatment led to increased expression of selected HIF-related genes, while GN44028 lacked clear selectivity over non-malignant cells.
Conclusions:
- Nuclear HIF activation is linked to aggressive ccRCC biology.
- Further investigation into broader HIF pathway modulation is warranted for ccRCC treatment.
- Clinical findings are exploratory, and therapeutic selectivity and translational relevance of HIF inhibitors require further establishment.
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