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

Abstract

BackgroundKnowledge 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.MethodsTo 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.ResultsIn 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.ConclusionBased 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.

Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells

“…An early study, focusing on the expression of 3-MST in cancer cells was performed by Wrobel's group and was found to detect significant 3-MST expression and enzymatic activity in several different human neoplastic cell lines (including U373 astrocytoma, SH-SY5Y neuroblastoma, and the melanoma lines A375 and WM35) [6,21]. There are many additional cancer cell lines where the presence of 3-MST has been demonstrated; these include the glioblastoma-astrocytoma cell line U-87 [6], the hepatoma lines Hepa1c17 and HepG2 [22,23], various human colon cancer cell lines (HCT116, HT-29, LoVo,) [24][25][26], the human lung adenocarcinoma line A549 [27,28], the RCC4 renal cell carcinoma line [29], several different urothelial cancer cell lines [30,31] and several melanoma cell lines (PES 43, A375, Sk-Mel-5, Sk-Mel-28) [32]. The results of the current study add the murine colon cancer cell line CT26 to the list of tumor cells that exhibit high 3-MST expression.…”

Release of reactive selenium species from phthalic selenoanhydride in the presence of hydrogen sulfide and glutathione with implications for cancer research

“…[38][39][40][41][42] It has both pro-and anticancer effects depending on the cell type, concentration and interaction with other cellular molecules. [43][44][45] Glutathione (GSH) is another intracellular natural antioxidant, which has many biological roles including modulation of cellular redox homeostasis and protection against reactive oxygen and nitrogen species. 46,47 As has been determined in previous work, sodium selenite (Na 2 SeO 3 ) and selenium tetrachloride (SeCl 4 ) have the ability to interact separately with GSH and H 2 S. This fact could be behind their biological effects.…”

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

“…CBS was shown to be localized mainly in cytosol and to a lesser extend in mitochondria [18,19]. However, few years ago we have shown a speci c CBS pattern by immuno uorescent staining [8], similar to staining of cytoskeleton. Thus, we built up a hypothesis that CBS might bind to cytoskeletal proteins.…”

“…This enzyme was increased also in thyroid malignancies [10]. However, decrease in CBS expression was determined in clear cell renal cell carcinoma that is spontaneously hypoxic, compared to matched controls and this decrease was dependent on the grade of tumor [8]. Question remains, whether higher CBS levels in tumors could be due to lower amount of H 2 S in tumor, as it was suggested by Dongsoo et al [11].…”

“…Expression of the CBS was studied in several types of tumors. Increased levels of CBS were detected in colon/colorectal cancer [6,7], papillary carcinoma [8], breast and also in prostate cancer [9]. This enzyme was increased also in thyroid malignancies [10].…”