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Roles of N-methyl-d-aspartate receptors and d-amino acids in cancer cell viability

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Abstract

N-methyl-d-aspartate (NMDA) receptors, which are widely present in the central nervous system, have also been found to be up-regulated in a variety of cancer cells and tumors and they can play active roles in cancer cell growth regulation. NMDA receptor antagonists have been found to affect cancer cell viability and interfere with tumor growth. Moreover, cancer cells also have been shown to have elevated levels of some d-amino acids. Two human skin cell lines: Hs 895.T skin cancer and Hs 895.Sk skin normal cells were investigated. They were derived from the same patient to provide tumor and normal counterparts for comparative studies. The expression of specific NMDA receptors was confirmed for the first time in both skin cell lines. Dizocilpine (MK-801) and memantine, NMDA receptor channel blockers, were found to inhibit the growth of human skin cells by reducing or stopping NMDA receptor activity. Addition of d-Ser, d-Ala, or d-Asp, however, significantly reversed the antiproliferative effect on the human skin cells triggered by MK-801 or memantine. Even more interesting was the finding that the specific intracellular composition of a few relatively uncommon amino acids was selectively elevated in skin cancer cells when exposed to MK-801. It appears that a few specific and upregulated d-amino acids can reverse the drug-induced antiproliferative effect in skin cancer cells via the reactivation of NMDA receptors. This study provides a possible innovative anticancer therapy by acting on the d-amino acid pathway in cancer cells either blocking or activating their regulatory enzymes.

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Data availability

The datasets used for the current study are available from the corresponding author on reasonable request.

Abbreviations

NMDA receptors:

N-methyl-d-aspartate receptors

MK-801:

Dizocilpine

CNS:

Central nervous system

DAAO:

d-amino acid oxidase

AD:

Alzheimer’s disease

AQC:

6-Aminoquinolyl-N-hydroxysuccinimide carbamate

HPLC–MS:

High performance liquid chromatography-mass spectrometry

BCA:

Bicinchoninic acid

DMEM:

Dulbecco’s Modified Eagle Medium

qRT-PCR:

Quantitative real-time polymerase chain reaction

PVDF:

Polyvinylidene fluoride

FBS:

Fetal bovine serum

ERK:

Extracellular signal-regulated kinase

SEM:

Standard error of mean

Akt:

Protein kinase B

PCNA:

Proliferating cell nuclear antigen

Aurora B:

Aurora B kinase

P450:

Cytochrome P450

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Acknowledgements

AZYP, LLC, is acknowledged for assistance and technical support for HPLC chiral column technology. We would also like to thank the Shimadzu Center for Advanced Analytical Chemistry for the use of the Shimadzu instrument (LCMS-8040).

Funding

This work was supported by the Robert A. Welch Foundation (Y0026) for DWA and (Y-1933–20170325) for FMM.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX, 76019, USA

    Siqi Du, Yu-Sheng Sung, Michael Wey, Yadi Wang, Nagham Alatrash, Frederick M. MacDonnell & Daniel W. Armstrong

  2. Institut des Sciences Analytiques, CNRS, Université de Lyon 1, 5 rue de la Doua, 69100, Villeurbanne, France

    Alain Berthod

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Contributions

DWA devised this project. SD, YS, MW, YW, and DWA designed the experiments, analyzed the data, and performed most of the experiments. NA and MW contributed to the cell culture and cell viability study. SD, MW, AB, and DWA contributed to the writing of the manuscript. All authors discussed the results and reviewed the manuscript.

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Correspondence to Daniel W. Armstrong.

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11033_2020_5733_MOESM1_ESM.pdf

Supplementary file3 (PDF 74 kb). Fig. S1. HPLC-MS/MS chromatograms of the separation of D- and L-amino acids. Peak identities for the separation of Leu, Ile, and Hyp: 1. D-allo-Ile, 2. D-Leu, 3. trans-L-Hyp, 4.cis- L-Hyp, 5.cis- D-Hyp, 6. L-leu, 7. L-Ile, 8. L-allo-Ile. Peak identities for the separation of Val and Nva: 1. D-Nva, 2. D-Val, 3. L-Nva, 4. L-Val

11033_2020_5733_MOESM2_ESM.tif

Supplementary file1 (TIF 205 kb). Fig. S2. The effect of D-amino acids addition (700 µM) on (A) Hs 895.T skin cancer, and (B) Hs 895.Sk skin normal cell growth

Supplementary file2 (PDF 88 kb)

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Du, S., Sung, YS., Wey, M. et al. Roles of N-methyl-d-aspartate receptors and d-amino acids in cancer cell viability. Mol Biol Rep 47, 6749–6758 (2020). https://doi.org/10.1007/s11033-020-05733-8

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