Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Hashemian Nik, D.
Role of the gluconeogenesis enzyme PCK2 in anchorage-independent growth of lung cancer cells
Humanmedizin; [ Diplomarbeit ] Medizinische Universitaet Graz; 2022. pp. 62
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Blümel Gabriele Agnes
- Leithner Katharina
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- Abstract:
- The gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 2 (PCK2) recently shifted into focus of metabolomic studies, possibly opening the door to new anti-tumor strategies. Previous studies have shown promising effects of PCK2 silencing in attached non-small cell like cancer (NSCLC) cells under low glucose environments. The main objective of this study was to establish a suitable experimental model to investigate the effect of PCK2 under the special circumstance of detachment from the extracellular matrix (ECM). ECM detachment is crucial in the formation of metastasis when tumor cells break free from their surrounding tissue. In our setting, H23 NSCLC PCK2 expressing or silenced cells were cultured in ultra-low attachment plates under high and low glucose conditions, with or without serum supplementation. As methods, we used fluorescent cell scanning analysis (FACS), automated cell counting (ACC) and proliferation assay. We also compared PCK2 expression differences between attached and detached cells using Western Blot. PCK2 was expressed in H23 cells under all conditions, no significant differences were found. PCK2 silencing by two pools of siRNAs significantly reduced viable cell number in high glucose, serum containing media (p = 0.0031 and p = 0.041) and low glucose, serum free media (p = 0.0045). Overall, the absence of serum led to a clear reduction in viable cell number and proliferation in non-adherent cells. An increased expression of fatty acid synthase under serum-free conditions suggests that the cells might lack serum lipids under these conditions. In conclusion, we found a suitable model to test PCK2 effects on cell survival in anchorage-independent growth conditions. The results suggest that PCK2 might play a role in growth of cancer cells under these conditions, however further studies are needed to clarify whether inhibition of PCK2 in tumors could potentially interfere with metastasis formation.