Quantification of intermediates of the methionine and polyamine metabolism by liquid chromatography–tandem mass spectrometry in cultured tumor cells and liver biopsies

Stevens, Axel Peter; Dettmer, Katja; Kirovski, Georgi; Samejima, Keijiro; Hellerbrand, Claus; Bosserhoff, Anja‐Katrin; Oefner, Peter J.

doi:10.1016/j.chroma.2010.01.025

Cited by 43 publications

(38 citation statements)

References 25 publications

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“…Unlike previous analyzing methods of polyamine, the process of separation and quantitative was carried out without any derivatization and tedious sample preparation in this study, which keep the chromatography as simple as possible. Moreover, the developed method was successfully applied to the quantification of nine polyamines, four polyamine biosynthetic precursor and one catabolite in human plasma and urine, not only in the stage of cell culture medium [24], which benefit further cancer research in both laboratorial and clinical field.…”

Section: Introductionmentioning

confidence: 99%

Determination of polyamine metabolome in plasma and urine by ultrahigh performance liquid chromatography–tandem mass spectrometry method: Application to identify potential markers for human hepatic cancer

Liu

et al. 2013

Analytica Chimica Acta

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Section: Introductionmentioning

confidence: 99%

Determination of polyamine metabolome in plasma and urine by ultrahigh performance liquid chromatography–tandem mass spectrometry method: Application to identify potential markers for human hepatic cancer

Liu

et al. 2013

Analytica Chimica Acta

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“…Accordingly, trace detection by mass spectrometry had been developed for the polyamines determination. Q‐time of flight (Q‐TOF) mass spectrometry possesses high resolution and accurate detection, and which is currently a useful tool analyzing compounds in complex matrix . The coupling of high‐performance liquid chromatography (HPLC) and Q‐TOF mass spectrometry has advantages over gas chromatograph and HPLC‐coupled fluorescence detector for high‐resolution and narrow extracted‐ion windows.…”

Section: Introductionmentioning

confidence: 99%

Determination of polyamines in human plasma by high‐performance liquid chromatography coupled with Q‐TOF mass spectrometry

Liu

Jia

et al. 2012

J. Mass. Spectrom.

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A high-performance liquid chromatography coupled with Q-time of flight mass spectrometry (HPLC/Q-TOF MS) method was developed and validated for the determination of 1, 3-diaminopropane, putrescine, cadaverine, spermidine and spermine in human plasma. The plasma samples were first pretreated by 10% HClO(4) and then derived by benzoyl chloride with 1, 6-diaminohexane as internal standard. The derived polyamines were separated on a C(18) column using a gradient program. The detection was performed on a Q-TOF MS by positive ionization mode. Calibration curve for each polyamine was obtained in the concentration range of 0.4 ~ 200.0 ng • ml(-1), with limit of detection of 0.02 ~ 0.1 ng • ml(-1). The intra- and inter-day RSD for all polyamines were 2.5-14.0% and 2.9 ~ 13.4%, respectively. The method was applied to determine the polyamines in human plasma from cancer patients and healthy volunteers. Results showed that the mean levels of polyamines in the plasma of cancer patients were higher than that of healthy volunteers, which suggested that the plasma polyamines could be employed as cancer diagnostic indicators in clinical testing.

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“…Samples were prepared as described by Stevens et al 81,82 Briefly, for determination of extracellular MTA, 200 mL of cell culture medium were transferred to 2-mL Eppendorf tubes (Eppendorf) and spiked with 10 mL stable isotope labeled internal standard prior to protein precipitation with MeOH/0.1 M acetic acid (80:20, v/v). After centrifugation, the supernatant was transferred to a 1.5-mL glass vial (Fisher Scientific).…”

Section: Mta Extraction and Analysis By Lc-esi-ms/msmentioning

confidence: 99%

“…Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) was carried out according to Stevens et al 82 on an Agilent 1200 SL HPLC system (B€ oblingen, Germany) coupled to a 4000 QTRAP mass spectrometer (AB Sciex, Darmstadt, Germany). An Atlantis T3 3 mm (2.1 mm i.d.x 150 mm) reversed phase column (Waters, Eschborn, Germany) served as stationary phase, while the mobile phase contained 0.1% acetic acid and 0.025% heptafluorobutyric acid.…”

Section: Mta Extraction and Analysis By Lc-esi-ms/msmentioning

confidence: 99%

Suppressive effects of tumor cell-derived 5′-deoxy-5′-methylthioadenosine on human T cells

et al. 2016

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The immunosuppressive tumor microenvironment represents one of the main obstacles for immunotherapy of cancer. The tumor milieu is among others shaped by tumor metabolites such as 5 0 -deoxy-5 0 -methylthioadenosine (MTA). Increased intratumoral MTA levels result from a lack of the MTAcatabolizing enzyme methylthioadenosine phosphorylase (MTAP) in tumor cells and are found in various tumor entities. Here, we demonstrate that MTA suppresses proliferation, activation, differentiation, and effector function of antigen-specific T cells without eliciting cell death. Conversely, if MTA is added to highly activated T cells, MTA exerts cytotoxic effects on T cells. We identified the Akt pathway, a critical signal pathway for T cell activation, as a target of MTA, while, for example, p38 remained unaffected. Next, we provide evidence that MTA exerts its immunosuppressive effects by interfering with protein methylation in T cells. To confirm the relevance of the suppressive effects of exogenously added MTA on human T cells, we used an MTAP-deficient tumor cell-line that was stably transfected with the MTAPcoding sequence. We observed that T cells stimulated with MTAP-transfected tumor cells revealed a higher proliferative capacity compared to T cells stimulated with Mock-transfected cells. In conclusion, our findings reveal a novel immune evasion strategy of human tumor cells that could be of interest for therapeutic targeting.

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Quantification of intermediates of the methionine and polyamine metabolism by liquid chromatography–tandem mass spectrometry in cultured tumor cells and liver biopsies

Cited by 43 publications

References 25 publications

Determination of polyamine metabolome in plasma and urine by ultrahigh performance liquid chromatography–tandem mass spectrometry method: Application to identify potential markers for human hepatic cancer

Determination of polyamine metabolome in plasma and urine by ultrahigh performance liquid chromatography–tandem mass spectrometry method: Application to identify potential markers for human hepatic cancer

Determination of polyamines in human plasma by high‐performance liquid chromatography coupled with Q‐TOF mass spectrometry

Suppressive effects of tumor cell-derived 5′-deoxy-5′-methylthioadenosine on human T cells

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