The impact of hypoxia on the cellular phenotype and proteome of KGN cells

Human KGN cells are a widely studied cellular model for human granulosa cell tumors (GCTs), as well as for non-tumorous human granulosa cells. Such studies are typically performed at oxygen (O 2 ) levels, which are equivalent to the atmospheric concentration, for practical reasons. However, in most tissues of the human body, as well as in tumors, much lower O 2 levels exist. Hypoxic conditions can regulate the phenotype and the behavior of cells in a cell-type-specific way. However, how they may impact KGN cells, was not well known. The aim of this study was therefore to evaluate the consequences of a defined hypoxic condition (1% O 2 ; 10 days) in KGN cells. Compared to cells cultured under atmospheric conditions, hypoxia reduced the numbers and increased the size of KGN cells, but did not alter cell velocity in cell migration studies. Hypoxia affected the cellular proteome of KGN cells (57 more and 75 less abundant proteins). The associated pathways indicate, among others, metabolic adaptations and mitochondrial changes evoked by hypoxia. The KGN responses may be related, in part, to the increased nuclear expression of hypoxia inducible transcription factor 1α (HIF1α). The results provide a detailed picture of hypoxia-induced changes in KGNs and are a resource for future studies. If transferable to the in situ -situation, they shed light on how variable O 2 levels within the GCT microenvironment may regulate tumor cells and thereby may contribute to tumor cell heterogeneity. Lay summary Human cells can be isolated from the body and can be kept alive in incubators, in which, mainly for practical reasons, the oxygen (O 2 ) levels typically correspond to the surrounding atmosphere. This does, however, not reflect the condition within the human body, where much lower O 2 levels exist. This study shows the importance of low O 2 levels on the cellular make-up and behavior of KGN cells, which are accepted models for a group of ovarian tumors (granulosa cell tumors), as well as normal non-tumorous granulosa cells (from which they originate). We found that when KGN cells were exposed to low O 2 levels, cell numbers and cell size, as well as cellular proteins and RNA changed. The experiments, revealing adaptations associated with hypoxia, may contribute to a better understanding of the normal ovary and ovarian tumors.