Detection of Morphology, Gene Expression and Protein Alterations in Mesenchymal Stem Cells from Bone Marrow after Exposure to Aflatoxin B1
Keywords:
gene mutation, molecular biomarkers, bone marrow, mesenchymal stem cell, proteome patternAbstract
Cancer is a major public-health problem worldwide. The treatments and diagnostic techniques for early-stage cancer remain limited. Understanding the mechanisms of carcinogenesis can assist cancer control. The use of human mesenchymal stem cells derived from bone marrow as a model to study environmental carcinogenesis is of interest and has never been undertaken in Thailand. Therefore, this study aimed to detect morphology, gene expression, and protein alterations, in mesenchymal stem cells from bone marrow after exposure to carcinogenic aflatoxin B1 (AFB1). In this study, concentrations of AFB1 at 25 µM, 50 µM, and 100 µM, were applied to stem cells. At 8, 16, and 24 hours post-exposure, we observed changes in morphology, number of cells, Bmi-1 gene expression relative to GAPDH by real-time PCR, and detected proteome pattern by MALDITOF/TOF mass spectrometer. The findings showed that the expression of the Bmi-1 oncogene, which controls cell division, increased 2-fold compared with normal cells (2−∆∆CT = 2.07) after exposure to AFB1 at 50 µM AFB1 within 24 hours, indicating that the mesenchymal stem cells from bone marrow after exposure to carcinogenic AFB1 for 24 hours were being gradually changed into cancer cells, but not yet completely converted to cancer cells. Therefore, alterations in morphology and number of stem cells were not detected. Proteome pattern detection showed that the protein with a molecular weight of 2423.79 daltons was upregulated such that the cells were going to become cancerous, and the protein with a molecular weight of 3174.93 daltons was downregulated, such that the cells were going to become cancerous. This study demonstrated that we can use human mesenchymal stem cells from bone marrow as a model to study the mechanisms of carcinogenesis instead of laboratory animals. In addition, diagnostic and prognostic genetic and protein biomarkers may be discovered, leading to the early detection of cancers in the future.
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