Inhibition of cytosolic and mitochondrial creatine kinase by siRNA in HaCaT- and HeLaS3-cells affects cell viability and mitochondrial morphology
| Autor: | Franz Staeb, Horst Wenck, Melanie Schmidt, Vivienne Welge, Holger Lenz, Uwe Schlattner, Thomas Kueper, Hans-Peter Elsässer, Theo Wallimann, Klaus-Peter Wittern, Thomas Blatt |
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| Přispěvatelé: | R&D, Beiersdorf AG, Laboratoire de bioénergétique fondamentale et appliquée (LBFA), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Biology, Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)-Institute of Cell Biology, Department of Cytobiology and Cytopathology, Philipps Universität Marburg, Hamant, Sarah, Philipps Universität Marburg = Philipps University of Marburg |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
Keratinocytes
Phosphocreatine Cell Survival MESH: Mitochondria Clinical Biochemistry Creatine Kinase Mitochondrial Form Mitochondrion MESH: Phosphocreatine 03 medical and health sciences Cytosol 0302 clinical medicine MESH: Cytosol Creatine Kinase BB Form MESH: RNA Small Interfering [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology Humans MESH: Creatine Kinase Mitochondrial Form [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology RNA Messenger Viability assay RNA Small Interfering Inner mitochondrial membrane Molecular Biology 030304 developmental biology MESH: RNA Messenger 0303 health sciences MESH: Humans biology Cell growth Cell Biology General Medicine MESH: Creatine Kinase BB Form Molecular biology MESH: Gene Expression Regulation MESH: Keratinocytes Mitochondria Cell biology Isoenzymes MESH: Hela Cells HaCaT Gene Expression Regulation MESH: Cell Survival Cancer cell biology.protein MESH: Isoenzymes Creatine kinase 030217 neurology & neurosurgery HeLa Cells |
| Zdroj: | Molecular and Cellular Biochemistry Molecular and Cellular Biochemistry, Springer Verlag, 2007, 306 (1-2), pp.153-62. ⟨10.1007/s11010-007-9565-8⟩ |
| ISSN: | 0300-8177 1573-4919 |
| DOI: | 10.1007/s11010-007-9565-8⟩ |
| Popis: | International audience; The creatine kinase (CK) system is essential for cellular energetics in tissues or cells with high and fluctuating energy requirements. Creatine itself is known to protect cells from stress-induced injury. By using an siRNA approach to silence the CK isoenzymes in human keratinocyte HaCaT cells, expressing low levels of cytoplasmic CK and high levels of mitochondrial CK, as well as HeLa cancer cells, expressing high levels of cytoplasmic CK and low levels of mitochondrial CK, we successfully lowered the respective CK expression levels and studied the effects of either abolishing cytosolic brain-type BB-CK or ubiquitous mitochondrial uMi-CK in these cells. In both cell lines, targeting the dominant CK isoform by the respective siRNAs had the strongest effect on overall CK activity. However, irrespective of the expression level in both cell lines, inhibition of the mitochondrial CK isoform generally caused the strongest decline in cell viability and cell proliferation. These findings are congruent with electron microscopic data showing substantial alteration of mitochondrial morphology as well as mitochondrial membrane topology after targeting uMi-CK in both cell lines. Only for the rate of apoptosis, it was the least expressed CK present in each of the cell lines whose inhibition led to the highest proportion of apoptotic cells, i.e., downregulation of uMi-CK in case of HeLaS3 and BB-CK in case of HaCaT cells. We conclude from these data that a major phenotype is linked to reduction of mitochondrial CK alone or in combination with cytosolic CK, and that this effect is independent of the relative expression levels of Mi-CK in the cell type considered. The mitochondrial CK isoform appears to play the most crucial role in maintaining cell viability by stabilizing contact sites between inner and outer mitochondrial membranes and maintaining local metabolite channeling, thus avoiding transition pore opening which eventually results in activation of caspase cell-death pathways. |
| Databáze: | OpenAIRE |
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