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Evaluation of lactase dehydrogenase activity as a marker of carbohydrate metabolism in melanoma cells line Me45 under the influence of cisplatin

Renata M. Polaniak 1Małgorzata T. Latocha 2Rafal J. Buldak 3Ewa Romuk 1Mateusz Grajek 4Ryszard Szkilnik 5Ewa Birkner 1

1. Department of General Biochemistry, Faculty of Medicine in Zabrze, Silesian Medical University, Jordana 19, Zabrze 41-808, Poland
2. Department of Cell Biology, Medical University of Silesia (SUM), Jedności 8, Sosnowiec 41-200, Poland
3. Department of Physiology, Medical University of Silesia, H. Jordana 38, Zabrze 41-808, Poland
4. Department of Dietetics, Medical Uniwersity of Silesia, H.Jordana 19, Zabrze 41-808, Poland
5. Department of Basic Medical Sciences, Medical Universityof Silesia, Piekarska18, Bytom 41-902, Poland


Cisplatin - CPL (cis-diaminodichloroplatin), a platinum derivative, belongs to the group of alkylating medicines. The mechanism of action of alkylating agents is based on damaging the biological activity of the deoxyribonucleic acid, DNA, through the formation of crosslinks between adjacent DNA strands and within the same strand. The formation of such strands prevents DNA replication and cell division. It also affects the metabolic processes and can trigger cell apoptosis. The aim of the study was to evaluate the activity of lactase dehydrogenase, LDH [EC] in cell supernatants as the survival indicator of melanoma cells line Me45 exposed to cisplatin. Lactate dehydrogenase is a marker of malignant diseases, cardiac and liver diseases. The metabolism of a cancer cell is based on the process of glycolysis. LDH is an enzyme that catalyzes the last step of glycolysis, i.e. the oxidation of pyruvic acid to lactic acid, so the activity of the total LDH may indicate the degree of cancer cell damage. Cultures were grown on the human melanoma Me45 cell line. The culture medium was Dulbeco's with L-glutamine, fetal bovine serum enhanced. Breeding was conducted under standard conditions of temperature and carbon dioxide-enriched atmosphere. Cells were treated with three concentrations of cisplatin: C1 = 0.125 mg/ml; C2 = 0.25 mg/ml; C3 = 0.5 mg/ml, the control group were cells with no cytostatic in the culture medium. The cell supernatant was collected after 24-hour exposure and lactate dehydrogenase activity was determined. The research found that the activity of the enzyme in the supernatants from the studied groups decreased compared to the control group. The results suggest that cisplatin, depending on the applied cytostatic dose, acts as a brake on the process of cell glycolysis, as shown by the observation of a decrease in LDH activity in cell supernatants of Me45 melanoma line treated with chemotherapy in in-vitro conditions.


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Submitted: 2014-03-15 17:55
Revised:   2014-05-02 17:33