Proteome-wide Identification of New Molecular Targets Affected by Methotrexate in Acute Promyelocytic Leukaemia Cell Line

摘要: Methotrexate (MTX) was first introduced as a cytotoxic agent that inhibits nucleotide biosynthesis in various cancer disorders. Accumulating evidences suggest MTX inhibits the proliferation of malignant cells by inhibiting 5-aminoimidazole-4-carbox-amide ribonucleotide transformylase, isoprenylcysteine carboxyl methyltransferase and NF-kappaB transcription factor. These observations indicate could have additional molecular targets are therefore unappreciated. To get insights into the complex mechanisms MTX induced apoptosis acute promyelocytic leukemia (HL 60), we conducted an investigation incorporating cysteine labeled differential gel electrophoresis combined with mass spectrometry. Initial experimental analysis revealed 24 proteins were differentially expressed (p > 0.05) HL 60 cell proteome after addition 2.5 µM for 72 hours. The majority of MTX induced proteins ascribed to endoplasmic reticulum (ER) chaperones, glycolytic enzymes mitochondrial transmembrane electron transport system (MTETS). In particular, we noted three structural alpha4, alpha5, alpha7; a non-catalytically beta3 two 26S regulatory proteasome subunits were significantly down regulated treated cell. We further showed induces ER chaperones, suppresses NF-kappaB subunit p65, disturbs potential (Aom) generates reactive oxygen species (ROS) a time dependent manner. All together, our findings revealed alter level variety involved in associated formation, ER stress, MTETS. Their identification as may provide new impulse understanding apoptotic activities leukaemia cell line.