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.

Department of Molecular Biology Faculty of Natural Sciences Comenius University Bratislava Slovakia

Department of Physiology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Urology with Kidney Transplant Center University Hospital Faculty of Medicine Bratislava Slovakia

Institute of Clinical and Translational Research Biomedical Research Center SAS Bratislava Slovakia

Zobrazit více v PubMed

Ciocci M, Iorio E, Carotenuto F, Khashoggi HA, Nanni F, Melino S. H2S-releasing nanoemulsions: a new formulation to inhibit tumor cells proliferation and improve tissue repair. Oncotarget. 2016;7:84338–84358. doi: 10.18632/oncotarget.12609. PubMed DOI PMC

Hudecova S, Markova J, Simko V, Csaderova L, Stracina T, Sirova M, et al. Sulforaphane-induced apoptosis involves the type 1 IP3 receptor. Oncotarget. 2016;7:61403–61418. doi: 10.18632/oncotarget.8968. PubMed DOI PMC

Wei Z, Shan Y, Tao L, Liu Y, Zhu Z, Liu Z, et al. Diallyl trisulfides, a natural histone deacetylase inhibitor, attenuate HIF-1α synthesis and decreases breast cancer metastasis. Mol Carcinog. 2017; 10.1002/mc.22686. PubMed

Wu D, Si W, Wang M, Lv S, Ji A, Li Y. Hydrogen sulfide in cancer: friend or foe? Nitric Oxide. 2015;50:38–45. doi: 10.1016/j.niox.2015.08.004. PubMed DOI

Untereiner AA, Oláh G, Módis K, Hellmich MR, Szabo C. H2S-induced S-sulfhydration of lactate dehydrogenase a (LDHA) stimulates cellular bioenergetics in HCT116 colon cancer cells. Biochem Pharmacol. 2017;136:86–98. doi: 10.1016/j.bcp.2017.03.025. PubMed DOI PMC

Bhattacharyya S, Saha S, Giri K, Lanza IR, Nair KS, Jennings NB, et al. Cystathionine betasynthase (CBS) contributes to advanced ovarian cancer progression and drug resistance. PLoS One. 2013;8:e79167. doi: 10.1371/journal.pone.0079167. PubMed DOI PMC

Sen S, Kawahara B, Gupta D, Tsai R, Khachatryan M, Roy Chowdhuri S, et al. Role of cystathionine β-synthase in human breast cancer. Free Radic Biol Med. 2015;86:228–238. doi: 10.1016/j.freeradbiomed.2015.05.024. PubMed DOI

Kim J, Hong SJ, Park JH, Park SY, Kim SW, Cho EY, et al. Expression of cystathionine beta-synthase is downregulated in hepatocellular carcinoma and associated with poor prognosis. Oncol Rep. 2009;21:1449–1454. PubMed

Zhao H, Li Q, Wang J, Su X, Ng KM, Qiu T, et al. Frequent epigenetic silencing of the folate-metabolising gene cystathionine-beta-synthase in gastrointestinal cancer. PLoS One. 2012;7:e49683. doi: 10.1371/journal.pone.0049683. PubMed DOI PMC

Guo H, Gai JW, Wang Y, Jin HF, Du JB, Jin J. Characterization of hydrogen sulfide and its synthases, cystathionine β-synthase and cystathionine γ-lyase, in human prostatic tissue and cells. Urology. 2012;79(483):e1–e5. PubMed

Liu M, Wu L, Montaut S, Yang G. Hydrogen sulfide signaling Axis as a target for prostate Cancer therapeutics. Prostate Cancer. 2016;2016:8108549. doi: 10.1155/2016/8108549. PubMed DOI PMC

Zhao K, Li S, Wu L, Lai C, Yang G. Hydrogen sulfide represses androgen receptor transactivation by targeting at the second zinc finger module. J Biol Chem. 2014;289:20824–20835. doi: 10.1074/jbc.M114.559518. PubMed DOI PMC

Sekiguchi F, Sekimoto T, Ogura A, Kawabata A. Endogenous hydrogen sulfide enhances cell proliferation of human gastric Cancer AGS cells. Biol Pharm Bull. 2016;39:887–890. doi: 10.1248/bpb.b15-01015. PubMed DOI

Panza E, De Cicco P, Armogida C, Scognamiglio G, Gigantino V, Botti G, et al. Role of the cystathionine γ lyase/hydrogen sulfide pathway in human melanoma progression. Pigment Cell Melanoma Res. 2015;28:61–72. doi: 10.1111/pcmr.12312. PubMed DOI

Cao X, Bian JS. The role of hydrogen sulfide in renal system. Front Pharmacol. 2016;7:385. doi: 10.3389/fphar.2016.00385. PubMed DOI PMC

Kovacs G, Akhtar M, Beckwith BJ, Bugert P, Cooper CS, Delahunt B, et al. The Heidelberg classification of renal cell tumours. J Pathol. 1997;183:131–133. doi: 10.1002/(SICI)1096-9896(199710)183:2<131::AID-PATH931>3.0.CO;2-G. PubMed DOI

Hu X, Li T, Bi S, Jin Z, Zhou G, Bai C, et al. Possible role of hydrogen sulfide on the preservation of donor rat hearts. Transplant Proc. 2007;39:3024–3029. doi: 10.1016/j.transproceed.2007.05.086. PubMed DOI

Taniguchi S, Kang L, Kimura T, Niki I. Hydrogen sulphide protects mouse pancreatic beta-cells from cell death induced by oxidative stress, but not by endoplasmic reticulum stress. Br J Pharmacol. 2011;162:1171–1178. doi: 10.1111/j.1476-5381.2010.01119.x. PubMed DOI PMC

Shackelford RE, Abdulsattar J, Wei EX, Shackelford RE, Abdulsattar J, Wei EX, et al. Increased nicotinamide Phosphoribosyltransferase and cystathionine-β-synthase in renal Oncocytomas, renal urothelial carcinoma, and renal clear cell carcinoma. Anticancer Res. 2017;37:3423–3427. PubMed PMC

Soltysova A, Breza J, Takacova M, Feruszova J, Hudecova S, Novotna B, et al. Deregulation of energetic metabolism in the clear cell carcinoma: a multiple pathway analysis based on microarray profiling. Int J Oncol. 2015;47:287–295. doi: 10.3892/ijo.2015.3014. PubMed DOI

Asimakopoulou A, Panapoulos P, Chasapis CT, Coletta C, Zhou Z, Cirino G, et al. Selectivity of commonly used pharmacological inhibitors for cystathionine β synthase (CBS) and cystathionine γ lyase (CSE) Br J Pharmacol. 2013;169:922–932. doi: 10.1111/bph.12171. PubMed DOI PMC

Takano Y, Echizen H, Hanaoka K. Fluorescent probes and selective inhibitors for biological studies of hydrogen sulfide- and polysulfide-mediated signaling. Antioxid Redox Signal. 2017;27:669–683. doi: 10.1089/ars.2017.7070. PubMed DOI PMC

Chao C, Zatarain JR, Ding Y, Coletta C, Mrazek AA, Druzhyna N, et al. Cystathionine-beta-synthase inhibition for colon cancer: enhancement of the efficacy of aminooxyacetic acid via the prodrug approach. Mol Med. 2016;22 10.2119/molmed. PubMed PMC

Takano N, Sarfraz Y, Gilkes DM, Chaturvedi P, Xiang L, Suematsu M, et al. Decreased expression of cystathionine β-synthase promotes glioma tumorigenesis. Mol Cancer Res. 2014;12:1398–1406. doi: 10.1158/1541-7786.MCR-14-0184. PubMed DOI PMC

Han SJ, Kim JI, Park JW, Park KM. Hydrogen sulfide accelerates the recovery of kidney tubules after renal ischemia/reperfusion injury. Nephrol Dial Transplant. 2015;30:1497–1506. doi: 10.1093/ndt/gfv226. PubMed DOI

De Cicco P, Panza E, Ercolano G, Armogida C, Sessa G, Pirozzi G, et al. ATB-346, a novel hydrogen sulfide-releasing anti-inflammatory drug, induces apoptosis of human melanoma cells and inhibits melanoma development in vivo. Pharmacol Res. 2016;114:67–73. doi: 10.1016/j.phrs.2016.10.019. PubMed DOI

Lencesova L, Vlcek M, Krizanova O, Hudecova S. Hypoxic conditions increases H2S-induced ER stress in A2870 cells. Mol Cell Biochem. 2016;414:67–76. doi: 10.1007/s11010-016-2659-4. PubMed DOI

Lu S, Chen L, Huang Q, Yang L, Du C, Li X, et al. Decomposition of ammonia and hydrogen sulfide sludge drying waste gas by a novel non-thermal plasma. Chemosphere. 2014;117:781–785. doi: 10.1016/j.chemosphere.2014.10.036. PubMed DOI

Markova J, Hudecova S, Soltysova A, Sirova M, Csaderova L, Lencesova L, et al. Sodium/calcium exchanger is upregulated by sulfide signaling, forms complex with the beta1 and beta3 but not beta2 adrenergic receptors, and induces apoptosis. Pflugers Arch. 2014;466:1329–1342. doi: 10.1007/s00424-013-1366-1. PubMed DOI

Misak A, Grman M, Bacova Z, Rezuchova I, Hudecova S, Ondriasova E, Krizanova O, Brezova V, Chovanec M, Ondrias K. Polysulfides and products of H2S/S-nitrosoglutathione in comparison to H2S, glutathione and antioxidant Trolox are potent scavengers of superoxide anion radical and produce hydroxyl radical by decomposition of H2O2. Nitric Oxide. 2018;76:136–151. doi: 10.1016/j.niox.2017.09.006. PubMed DOI

Cystathionine β-synthase affects organization of cytoskeleton and modulates carcinogenesis in colorectal carcinoma cells