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Endogenous H2S producing enzymes are involved in apoptosis induction in clear cell renal cell carcinoma

J. Breza, A. Soltysova, S. Hudecova, A. Penesova, I. Szadvari, P. Babula, B. Chovancova, L. Lencesova, O. Pos, J. Breza, K. Ondrias, O. Krizanova,

. 2018 ; 18 (1) : 591. [pub] 20180524

Language English Country England, Great Britain

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc19012652

BACKGROUND: Knowledge about the expression and thus a role of enzymes that produce endogenous H2S - cystathionine-β-synthase, cystathionine γ-lyase and mercaptopyruvate sulfurtransferase - in renal tumors is still controversial. In this study we aimed to determine the expression of these enzymes relatively to the expression in unaffected part of kidney from the same patient and to found relation of these changes to apoptosis. To evaluate patient's samples, microarray and immunohistochemistry was used. METHODS: To determine the physiological importance, we used RCC4 stable cell line derived from clear cell renal cell carcinoma, where apoptosis induction by a mixture of five chemotherapeutics with/without silencing of H2S-producing enzymes was detected. Immunofluorescence was used to determine each enzyme in the cells. RESULTS: In clear cell renal cell carcinomas, expression of H2S-producing enzymes was mostly decreased compared to a part of kidney that was distal from the tumor. To evaluate a potential role of H2S-producing enzymes in the apoptosis induction, we used RCC4 stable cell line. We have found that silencing of cystathionine-β-synthase and cystathionine γ-lyase prevented induction of apoptosis. Immunofluorescence staining clearly showed that these enzymes were upregulated during apoptosis in RCC4 cells. CONCLUSION: Based on these results we concluded that in clear cell renal cell carcinoma, reduced expression of the H2S-producing enzymes, mainly cystathionine γ-lyase, might contribute to a resistance to the induction of apoptosis. Increased production of the endogenous H2S, or donation from the external sources might be of a therapeutic importance in these tumors.

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$a BACKGROUND: Knowledge about the expression and thus a role of enzymes that produce endogenous H2S - cystathionine-β-synthase, cystathionine γ-lyase and mercaptopyruvate sulfurtransferase - in renal tumors is still controversial. In this study we aimed to determine the expression of these enzymes relatively to the expression in unaffected part of kidney from the same patient and to found relation of these changes to apoptosis. To evaluate patient's samples, microarray and immunohistochemistry was used. METHODS: To determine the physiological importance, we used RCC4 stable cell line derived from clear cell renal cell carcinoma, where apoptosis induction by a mixture of five chemotherapeutics with/without silencing of H2S-producing enzymes was detected. Immunofluorescence was used to determine each enzyme in the cells. RESULTS: In clear cell renal cell carcinomas, expression of H2S-producing enzymes was mostly decreased compared to a part of kidney that was distal from the tumor. To evaluate a potential role of H2S-producing enzymes in the apoptosis induction, we used RCC4 stable cell line. We have found that silencing of cystathionine-β-synthase and cystathionine γ-lyase prevented induction of apoptosis. Immunofluorescence staining clearly showed that these enzymes were upregulated during apoptosis in RCC4 cells. CONCLUSION: Based on these results we concluded that in clear cell renal cell carcinoma, reduced expression of the H2S-producing enzymes, mainly cystathionine γ-lyase, might contribute to a resistance to the induction of apoptosis. Increased production of the endogenous H2S, or donation from the external sources might be of a therapeutic importance in these tumors.
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$a Soltysova, Andrea $u Institute of Clinical and Translational Research, Biomedical Research Center, SAS, Bratislava, Slovakia. Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia.
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$a Hudecova, Sona $u Institute of Clinical and Translational Research, Biomedical Research Center, SAS, Bratislava, Slovakia.
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$a Szadvári, Ivan, $u Department of Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. $d 1985- $7 xx0304142
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$a Babula, Petr $u Department of Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
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$a Pos, Ondrej $u Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia.
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$a Ondrias, Karol $u Institute of Clinical and Translational Research, Biomedical Research Center, SAS, Bratislava, Slovakia.
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$a Krizanova, Olga $u Institute of Clinical and Translational Research, Biomedical Research Center, SAS, Bratislava, Slovakia. olga.krizanova@savba.sk. Department of Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. olga.krizanova@savba.sk.
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