Distinct von Hippel-Lindau gene and hypoxia-regulated alterations in gene and protein expression patterns of renal cell carcinoma and their effects on metabolism

Abstract

During the last decade the knowledge about the molecular mechanisms of the cellular adaption to hypoxia and the function of the “von Hippel Lindau” (VHL) protein in renal cell carcinoma (RCC) has increased, but there exists little information about the overlap and differences in gene/protein expression of both processes. Therefore the aim of this study was to dissect VHL- and hypoxia-regulated alterations in the metabolism of human RCC using ome-based strategies. The effect of the VHL- and hypoxia-regulated altered gene/protein expression pattern on the cellular metabolism was analyzed by determination of glucose uptake, lactate secretion, extracellular pH, lactate dehydrogenase activity, amino acid content and ATP levels. By employing VHL−/VHL+ RCC cells cultured under normoxic and hypoxic conditions, VHL-dependent, HIF-dependent as well as VHL-/HIF-independent alterations in the gene and protein expression patterns were identified and further validated in other RCC cell lines. The genes/proteins differentially expressed under these distinct conditions were mainly involved in the cellular metabolism, which was accompanied by an altered metabolism as well as changes in the abundance of amino acids in VHL-deficient cells. In conclusion, the study reveals similarities, but also differences in the genes and proteins controlled by VHL functionality and hypoxia thereby demonstrating differences in the metabolic switch of RCC under these conditions.

Capture‐C reveals preformed chromatin interactions between HIF ‐binding sites and distant promoters

“…Finally, we examined whether long‐term HIF stabilization resulting from VHL inactivation might alter cis‐ interactions with these HIF‐binding sites by comparing Capture‐C signals in VHL‐reconstituted (786‐O+VHL) and defective (786‐O‐VHL) 786‐O cells (Appendix Fig S4). For the large part, interaction frequencies were comparable in VHL‐reconstituted and VHL‐deficient cells, with the exception of reduced interaction with the promoter of TSC22D2 in VHL‐reconstituted cells versus VHL‐defective cells, which correlates with a lack of hypoxic inducibility of this VHL‐regulated gene in these cells . Thus, neither short‐term activation of HIF by hypoxia, nor its long‐term stabilization by VHL inactivation greatly alters chromatin interactions with HIF‐binding sites.…”

Systemic immune-inflammation index predicts the clinical outcome in patients with metastatic renal cell cancer treated with sunitinib

“…A predominant role in kidney cancer is played by inactivation of Von Hippen Lindau (VHL) tumor suppressor gene with consequent increased cellular amount of Hypoxia-Inducible Factor-1 alpha (HIF-1a) that cause abnormal cellular growth and angiogenesis [2–4]. Therefore inhibition of angiogenesis represents the mainstay of treatment of metastatic RCC [5, 6].…”

Deletion of Von Hippel–Lindau Interferes with Hyper Osmolality Induced Gene Expression and Induces an Unfavorable Gene Expression Pattern

“…HEK293T cells were obtained from the DSMZ-German Collection of Microorganisms and Cell Cultures and cultivated in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% serum (fetal calf serum (FCS)) and 1% penicillin/streptavidin. The 786-0 and ectopic VHL expressing 786-0-VHL cells were a kind gift of Prof. Barbara Seliger [47]. These cells were maintained in DMEM supplemented with 10% fetal calf serum (FCS), 2 mM glutamine, 1 mM pyruvate, and 1% penicillin/streptomycin.…”