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Effect of S-adenosyl-L-methionine (SAM), an allosteric activator of cystathionine-β-synthase (CBS) on colorectal cancer cell proliferation and bioenergetics in vitro.
Nitric Oxide. 2014 Sep 15; 41:146-56.NO

Abstract

Recent data show that colon cancer cells selectively overexpress cystathionine-β-synthase (CBS), which produces hydrogen sulfide (H2S), to maintain cellular bioenergetics, support tumor growth and stimulate angiogenesis and vasorelaxation in the tumor microenvironment. The purpose of the current study was to investigate the effect of the allosteric CBS activator S-adenosyl-L-methionine (SAM) on the proliferation and bioenergetics of the CBS-expressing colon cancer cell line HCT116. The non-transformed, non-tumorigenic colon epithelial cell line NCM356 was used as control. For assessment of cell proliferation, the xCELLigence system was used. Bioenergetic function was measured by Extracellular Flux Analysis. Experiments using human recombinant CBS or HCT116 homogenates complemented the cell-based studies. SAM markedly enhanced CBS-mediated H2S production in vitro, especially when a combination of cysteine and homocysteine was used as substrates. Addition of SAM (0.1-3 mM) to HCT116 cells induced a concentration-dependent increase H2S production. SAM exerted time- and concentration-dependent modulatory effects on cell proliferation. At 0.1-1 mM SAM increased HCT116 proliferation between 0 and 12 h, while the highest SAM concentration (3 mM) inhibited proliferation. Over a longer time period (12-24 h), only the lowest concentration of SAM used (0.1 mM) stimulated cell proliferation; higher SAM concentrations produced a concentration-dependent inhibition. The short-term stimulatory effects of SAM were attenuated by the CBS inhibitor aminooxyacetic acid (AOAA) or by stable silencing of CBS. In contrast, the inhibitory effects of SAM on cell proliferation was unaffected by CBS inhibition or CBS silencing. In contrast to HCT116 cells, the lower rate of proliferation of the low-CBS expressor NCM356 cells was unaffected by SAM. Short-term (1 h) exposure of HCT116 cells to SAM induced a concentration-dependent increase in oxygen consumption and bioenergetic function at 0.1-1 mM, while 3 mM was inhibitory. Longer-term (72 h) exposure of HCT116 cells to all concentrations of SAM tested suppressed mitochondrial oxygen consumption rate, cellular ATP content and cell viability. The stimulatory effect of SAM on bioenergetics was attenuated in cells with stable CBS silencing, while the inhibitory effects were unaffected. In NCM356 cells SAM exerted smaller effects on cellular bioenergetics than in HCT116 cells. We have also observed a downregulation of CBS in response to prolonged exposure of SAM both in HCT116 and NCM356 cells. Taken together, the results demonstrate that H2S production in HCT116 cells is stimulated by the allosteric CBS activator, SAM. At low-to intermediate levels and early time periods the resulting H2S serves as an endogenous cancer cell growth and bioenergetic factor. In contrast, the inhibition of cell proliferation and bioenergetic function by SAM does not appear to relate to adverse autocrine effects of H2S resulting from CBS over-stimulation but, rather to CBS-independent pharmacological effects.

Authors+Show Affiliations

Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA; Department of Pharmacology, University of Patras, Patras, Greece.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.Department of Surgery, University of Texas Medical Branch, Galveston, TX, USA.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA; Department of Pharmacology, University of Patras, Patras, Greece.Department of Surgery, University of Texas Medical Branch, Galveston, TX, USA.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA. Electronic address: szabocsaba@aol.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24667534

Citation

Módis, Katalin, et al. "Effect of S-adenosyl-L-methionine (SAM), an Allosteric Activator of Cystathionine-β-synthase (CBS) On Colorectal Cancer Cell Proliferation and Bioenergetics in Vitro." Nitric Oxide : Biology and Chemistry, vol. 41, 2014, pp. 146-56.
Módis K, Coletta C, Asimakopoulou A, et al. Effect of S-adenosyl-L-methionine (SAM), an allosteric activator of cystathionine-β-synthase (CBS) on colorectal cancer cell proliferation and bioenergetics in vitro. Nitric Oxide. 2014;41:146-56.
Módis, K., Coletta, C., Asimakopoulou, A., Szczesny, B., Chao, C., Papapetropoulos, A., Hellmich, M. R., & Szabo, C. (2014). Effect of S-adenosyl-L-methionine (SAM), an allosteric activator of cystathionine-β-synthase (CBS) on colorectal cancer cell proliferation and bioenergetics in vitro. Nitric Oxide : Biology and Chemistry, 41, 146-56. https://doi.org/10.1016/j.niox.2014.03.001
Módis K, et al. Effect of S-adenosyl-L-methionine (SAM), an Allosteric Activator of Cystathionine-β-synthase (CBS) On Colorectal Cancer Cell Proliferation and Bioenergetics in Vitro. Nitric Oxide. 2014 Sep 15;41:146-56. PubMed PMID: 24667534.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effect of S-adenosyl-L-methionine (SAM), an allosteric activator of cystathionine-β-synthase (CBS) on colorectal cancer cell proliferation and bioenergetics in vitro. AU - Módis,Katalin, AU - Coletta,Ciro, AU - Asimakopoulou,Antonia, AU - Szczesny,Bartosz, AU - Chao,Celia, AU - Papapetropoulos,Andreas, AU - Hellmich,Mark R, AU - Szabo,Csaba, Y1 - 2014/03/22/ PY - 2013/12/13/received PY - 2014/02/17/revised PY - 2014/03/06/accepted PY - 2014/3/27/entrez PY - 2014/3/29/pubmed PY - 2015/6/2/medline KW - Allosteric modulation KW - Bioenergetics KW - Colorectal cancer KW - Mitochondria KW - Proliferation SP - 146 EP - 56 JF - Nitric oxide : biology and chemistry JO - Nitric Oxide VL - 41 N2 - Recent data show that colon cancer cells selectively overexpress cystathionine-β-synthase (CBS), which produces hydrogen sulfide (H2S), to maintain cellular bioenergetics, support tumor growth and stimulate angiogenesis and vasorelaxation in the tumor microenvironment. The purpose of the current study was to investigate the effect of the allosteric CBS activator S-adenosyl-L-methionine (SAM) on the proliferation and bioenergetics of the CBS-expressing colon cancer cell line HCT116. The non-transformed, non-tumorigenic colon epithelial cell line NCM356 was used as control. For assessment of cell proliferation, the xCELLigence system was used. Bioenergetic function was measured by Extracellular Flux Analysis. Experiments using human recombinant CBS or HCT116 homogenates complemented the cell-based studies. SAM markedly enhanced CBS-mediated H2S production in vitro, especially when a combination of cysteine and homocysteine was used as substrates. Addition of SAM (0.1-3 mM) to HCT116 cells induced a concentration-dependent increase H2S production. SAM exerted time- and concentration-dependent modulatory effects on cell proliferation. At 0.1-1 mM SAM increased HCT116 proliferation between 0 and 12 h, while the highest SAM concentration (3 mM) inhibited proliferation. Over a longer time period (12-24 h), only the lowest concentration of SAM used (0.1 mM) stimulated cell proliferation; higher SAM concentrations produced a concentration-dependent inhibition. The short-term stimulatory effects of SAM were attenuated by the CBS inhibitor aminooxyacetic acid (AOAA) or by stable silencing of CBS. In contrast, the inhibitory effects of SAM on cell proliferation was unaffected by CBS inhibition or CBS silencing. In contrast to HCT116 cells, the lower rate of proliferation of the low-CBS expressor NCM356 cells was unaffected by SAM. Short-term (1 h) exposure of HCT116 cells to SAM induced a concentration-dependent increase in oxygen consumption and bioenergetic function at 0.1-1 mM, while 3 mM was inhibitory. Longer-term (72 h) exposure of HCT116 cells to all concentrations of SAM tested suppressed mitochondrial oxygen consumption rate, cellular ATP content and cell viability. The stimulatory effect of SAM on bioenergetics was attenuated in cells with stable CBS silencing, while the inhibitory effects were unaffected. In NCM356 cells SAM exerted smaller effects on cellular bioenergetics than in HCT116 cells. We have also observed a downregulation of CBS in response to prolonged exposure of SAM both in HCT116 and NCM356 cells. Taken together, the results demonstrate that H2S production in HCT116 cells is stimulated by the allosteric CBS activator, SAM. At low-to intermediate levels and early time periods the resulting H2S serves as an endogenous cancer cell growth and bioenergetic factor. In contrast, the inhibition of cell proliferation and bioenergetic function by SAM does not appear to relate to adverse autocrine effects of H2S resulting from CBS over-stimulation but, rather to CBS-independent pharmacological effects. SN - 1089-8611 UR - https://www.unboundmedicine.com/medline/citation/24667534/Effect_of_S_adenosyl_L_methionine__SAM__an_allosteric_activator_of_cystathionine_β_synthase__CBS__on_colorectal_cancer_cell_proliferation_and_bioenergetics_in_vitro_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1089-8603(14)00021-4 DB - PRIME DP - Unbound Medicine ER -