Reversed phase and cation exchange liquid chromatography with spectrophotometric and elemental/molecular mass spectrometric detection for S-adenosyl methionine/S-adenosyl homocysteine ratios as methylation index in cell cultures of ovarian cancer

González, Teresita Montero; Cinti, M.; Montes-Bayón, Marı́a; Campa, M.R. Fernández de la; Blanco-González, Elisa

doi:10.1016/j.chroma.2015.03.028

Cited by 10 publications

(6 citation statements)

References 32 publications

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“…SAM is converted by donation of a methyl group to S-adenosylhomocysteine (SAH) (Figure 5F), a methyltransferase inhibitor. Intracellular ratios of SAM to SAH have been shown to reflect cellular methylation potential, with low ratios predicting hypomethylation (Chen et al, 2010;Iglesias Gonzá lez et al, 2015). We observed a significantly reduced SAM to SAH ratio in E8 hPSCs, consistent with partial DNA hypomethylation (Figure 5G).…”

Section: The Epigenetic State Of E8 Hpscs Reflects Adaptations Toward Increased Lipid Biogenesissupporting

confidence: 51%

Lipid Deprivation Induces a Stable, Naive-to-Primed Intermediate State of Pluripotency in Human PSCs

et al. 2019

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Current challenges in capturing naive human pluripotent stem cells (hPSCs) suggest that the factors regulating human naive versus primed pluripotency remain incompletely defined. Here we demonstrate that the widely used Essential 8 minimal medium (E8) captures hPSCs at a naive-to-primed intermediate state of pluripotency expressing several naive-like developmental, bioenergetic, and epigenomic features despite providing primed-statesustaining growth factor conditions. Transcriptionally, E8 hPSCs are marked by activated lipid biosynthesis and suppressed MAPK/TGF-b gene expression, resulting in endogenous ERK inhibition. These features are dependent on lipidfree culture conditions and are lost upon lipid exposure, whereas short-term pharmacological ERK inhibition restores naive-to-primed intermediate traits even in the presence of lipids. Finally, we identify de novo lipogenesis as a common transcriptional signature of E8 hPSCs and the pre-implantation human epiblast in vivo. These findings implicate exogenous lipid availability in regulating human pluripotency and define E8 hPSCs as a stable, naive-to-primed intermediate (NPI) pluripotent state.

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Section: The Epigenetic State Of E8 Hpscs Reflects Adaptations Toward Increased Lipid Biogenesissupporting

confidence: 51%

Lipid Deprivation Induces a Stable, Naive-to-Primed Intermediate State of Pluripotency in Human PSCs

et al. 2019

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“…An improved method using a teicoplanin aglycone column separated the enantiomers using an ion-pairing reversed-phase mode and a low column temperature [71]. An HPLC-ESI-MS/MS method for the determination of SAM and SAH in cultures of ovarian cancer cells was developed [13]. LODs of approximately 0.5 ng/mL for both targeted analytes allowed the designed strategy to evaluate the effect of cisplatin on changes in the methylation index between epithelial ovarian cell lines sensitive to (A2780) and resistant to (A2780CIS) to this drug after exposure to cisplatin.…”

Section: Electrochemical Detectionmentioning

confidence: 99%

“…SAM and SAH therefore regulate the normal level of methylation in DNA, and deregulation of the methionine cycle has serious cellular consequences, resulting in disease. The ratio of SAM/SAH, called the "methylation index," may be a useful indicator of the methylation capacity of the cell [13].…”

Section: Introductionmentioning

confidence: 99%

HPLC Analysis of Homocysteine and Related Compounds

Wada¹,

Nakamura²,

Nakashima³

2018

Non-Proteinogenic Amino Acids

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Homocysteine (Hcy), a sulfur-containing amino acid, is a representative intermediate metabolite of methionine (Met) to cysteine (Cys) via several intermediates. An elevated level of Hcy in plasma plays an important role in diseases such as neural tube defects and Down syndrome. Homocystinuria is the most common inborn error of sulfur metabolism and is caused by mutations in the metabolic enzymes of Hcy. These errors can be caused by abnormal levels of Met metabolites and classified on the basis of plasma Met levels. Additionally, Hcy and related compounds such as glutathione play an important role in maintaining homeostasis. Therefore, the simultaneous determination of Hcy and/or related compounds is required for appropriate clinical management of several diseases. The sulfur-containing amino acids and their derivatives in biological samples are quantified sensitively using high-performance liquid chromatography methods coupled with various detection methods such as UV/Vis, fluorescence, chemiluminescence, electrochemical, mass spectrometry, and tandem mass spectrometry. In this chapter, we review recent advances in these analytical methods and their applications.

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“…Methionine (Met), an essential amino acid, was substituted for by its precursor homocysteine (Hcy) in the culture medium. , “Methionine dependency”, one of the metabolic characters of cancer cells, occurs in various human cancer cells, including breast, colon, lung, kidney, bladder, neuroglia, and melanin cells. , As Breillout and Kreis pointed out, the higher the metastatic potential of the cell line, the higher the concentration of Met required to maintain its proliferation. , Herein, a novel Met-conjugated fluorescent probe (Cy-Met) based on NIR fluorophore (IR-780) for tumor imaging was designed and synthesized. It exhibited a strong emission at 790 nm, which was qualified for penetration through the deep tissues.…”

Section: Introductionmentioning

confidence: 99%

“…23,24 "Methionine dependency", one of the metabolic characters of cancer cells, occurs in various human cancer cells, including breast, colon, lung, kidney, bladder, neuroglia, and melanin cells. 25,26 As Breillout and Kreis pointed out, the higher the metastatic potential of the cell line, the higher the concentration of Met required to maintain its proliferation. 27,28 Herein, a novel Met-conjugated fluorescent probe (Cy-Met) based on NIR fluorophore (IR-780) for tumor imaging was designed and synthesized.…”

Section: ■ Introductionmentioning

confidence: 99%

Methionine-Decorated Near Infrared Fluorescent Probe for Prolonged Tumor Imaging

et al. 2018

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Methionine (Met) is one of the essential amino acids of which the transport system L is overexpressed in various tumor cells. In this study, a near-infrared fluorescent dye (IR-780) and methionine were conjugated through a piperazin-polyamines linker to form Cy-Met. The successful synthesis of Cy-Met was validated by optical characterization, NMR, and MS spectra. The absorption peak of Cy-Met was at 680 nm, and the fluorescence peak was at 790 nm. The cytotoxicity assay and cell imaging studies indicated that Cy-Met had good biocompatibility and specific affinity to tumor cells. The dynamic distribution and clearance investigations showed that Cy-Met was eliminated by the liver-intestine pathway. Notably, Cy-Met displayed tumor-targeting ability in U87, H22, and EAC tumor-bearing mice with an evident long circulation time. The results implied that Cy-Met could act as a promising fluorescence probe specialized for long-term tumor monitoring.

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Reversed phase and cation exchange liquid chromatography with spectrophotometric and elemental/molecular mass spectrometric detection for S-adenosyl methionine/S-adenosyl homocysteine ratios as methylation index in cell cultures of ovarian cancer

Cited by 10 publications

References 32 publications

Lipid Deprivation Induces a Stable, Naive-to-Primed Intermediate State of Pluripotency in Human PSCs

Lipid Deprivation Induces a Stable, Naive-to-Primed Intermediate State of Pluripotency in Human PSCs

HPLC Analysis of Homocysteine and Related Compounds

Methionine-Decorated Near Infrared Fluorescent Probe for Prolonged Tumor Imaging

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