Endogenous H 2 S producing enzymes are involved in apoptosis induction in clear cell renal cell carcinoma

Abstract

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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