Fermented wheat germ extract induces apoptosis and downregulation of major histocompatibility complex class I proteins in tumor T and B cell lines.
The fermented wheat germ extract (code name: MSC, trade name: Avemar), with standardized benzoquinone content has been shown to inhibit tumor propagation and metastases formation in vivo. The aim of this study was to understand the molecular and cellular mechanisms of the anti-tumor effect of MSC. Therefore, we have designed in vitro model experiments using T and B tumor lymphocytic cell lines. Tyrosine phosphorylation of intracellular proteins and elevation of the intracellular Ca2+ concentration were examined using immunoblotting with anti-phosphotyrosine antibody and cytofluorimetry by means of Ca2+ sensitive fluorescence dyes, Fluo-3AM and FuraRed-AM, respectively. Apoptosis was measured with cytofluorimetry by staining the DNA with propidium iodide and detecting the cell population. The level of the cell surface MHC class I molecules was analysed with indirect immunofluorescence on cytofluorimeter using a monoclonal antibody to the non-polymorphic region of the human MHC class I. MSC stimulated tyrosine phosphorylation of intracellular proteins and the influx of extracellular Ca2+ resulted in elevation of intracellular Ca2+ concentration. Prominent apoptosis of 20-40% was detected upon 24 h of MSC treatment of the cell lines. As a result of the MSC treatment, the amount of the cell surface MHC class I proteins was downregulated by 70-85% compared to the non-stimulated control. MSC did not induce a similar degree of apoptosis in healthy peripheral blood mononuclear cells. Inhibition of the cellular tyrosine phosphatase activity or Ca2+ influx resulted in the opposite effect increasing or diminishing the Avemar induced apoptosis as well as the MHC class I downregulation, respectively. A benzoquinone component (2,6-dimethoxi-p-benzoquinone) in MSC induced similar apoptosis and downregulation of the MHC class I molecules in the tumor T and B cell lines to that of MSC. These results suggest that MSC acts on lymphoid tumor cells by reducing MHC class I expression and selectively promoting apoptosis of tumor cells on a tyrosine phosphorylation and Ca2+ influx dependent way. One of the components in MSC, 2,6-dimethoxi-p-benzoquinone was shown to be an important factor in MSC mediated cell response.
Fajka-Boja
Roberta
R
Lymphocyte Signal Transduction Laboratory, Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary.
Hidvégi
Maté
M
Shoenfeld
Yehuda
Y
Ion
Gabriela
G
Demydenko
Dmytro
D
Tömösközi-Farkas
Rita
R
Vizler
Csaba
C
Telekes
András
A
Resetar
Akos
A
Monostori
Eva
E
D017209:Apoptosis; D001402:B-Lymphocytes / Q000166:cytology*; D016227:Benzoquinones / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D019084:Fluorescent Antibody Technique, Indirect; D005805:Genes, MHC Class I / Q000235:genetics*; D006801:Humans; D019169:Jurkat Cells; D037102:Lectins / Q000494:pharmacology*; D017493:Leukocyte Common Antigens / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D010641:Phenotype; D010766:Phosphorylation; D010936:Plant Extracts / Q000494:pharmacology*; D037121:Plant Lectins; D013601:T-Lymphocytes / Q000166:cytology*; D013997:Time Factors; D014908:Triticum / Q000737:chemistry*; D014407:Tumor Cells, Cultured
C113915:Avemar; D016227:Benzoquinones; D037102:Lectins; D010936:Plant Extracts; D037121:Plant Lectins; C004532:quinone; D017493:Leukocyte Common Antigens; D002118:Calcium