[Expression of sperm-specific glyceraldehyde-3-phosphate dehydrogenase in melanoma cells changes their energy metabolism].

The somatic isoform of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC1.2.1.12) is involved in such crucial for cancer cells development pathways as induction of apoptosis and glycolytic regulation. At the same time, sperm-specific isoform (GAPDHS) does not exhibit all the same functions as somatic enzyme. The expression of sperm-specific GAPDH without N-terminal domain in some melanoma cells along with somatic isoenzyme, shown in our previous work, has led to the proposal of this unusual enzyme's possible role in regulation of cancer cells glycolysis. In the presented work we have tested production of GAPDHS in 13 additional melanoma cell lines by immunoblotting. We have also gathered data on energy metabolism in 5 selected cell lines by evaluation of glucose uptake and lactate production in differing conditions. We have demonstrated that in standard cultivation media glucose uptake by MelP cells, producing substantial amounts of GAPDHS protein was higher than in MelKor cells, producing lesser amounts of GAPDHS. All other analyzed cell lines that do not produce GAPDHS (MelMS, MelSi and Malme3M) had even a lower glucose uptake rate. Somaticheskaia forma glikoliticheskogo fermenta glitseral'degid-3-fosfatdegidrogenazy (GAFD; KF 1.2.1.12) vovlechena v takie vazhnye dlia razvitiia rakovykh kletok mekhanizmy kak induktsiia apoptoza i reguliatsiia glikoliza. V to zhe vremia, spermospetsifichnaia GAFD (GAFDs) ne obladaet vsem naborom funktsiĭ somaticheskogo fermenta. Obnaruzhennyĭ nami ranee fakt ékspressii v nekotorykh melanomnykh kletochnykh liniiakh spermospetsifichnoĭ GAFD bez N-kontsevogo domena nariadu s somaticheskoĭ formoĭ pozvolil predpolozhit' vozmozhnoe uchastie étoĭ neobychnoĭ formy v reguliatsii metabolizma rakovykh kletok. V dannoĭ rabote my protestirovali produktsiiu GAFDs metodom immunoblottinga v eshche 13 kletochnykh liniiakh melanomy. My takzhe poluchili dannye ob osnovnykh osobennostiakh énergeticheskogo obmena v vybrannykh piati kletochnykh liniiakh putem otsenki skorosti pogloshcheniia gliukozy i produktsii laktata v raznykh usloviiakh. Nami pokazano, chto v standartnykh usloviiakh kul'tivatsii pogloshchenie gliukozy v kletkakh MelP, produtsiruiushchikh bol'shie kolichestva GAFDs bylo vyshe, chem v MelKor, v kotorykh produktsiia GAFDs byla nizhe; v ostal'nykh issledovannykh liniiakh, ne produtsiruiushchikh GAFDs (MelMS, MelSi i Malme3M), skorost' pogloshcheniia gliukozy byla eshche nizhe.