AIM To explore the effect of branched chain amino acid transaminase 1 (BCAT1) on fibrosis related gene expression and phenotype during the transformation of fibroblasts into myofibroblasts.
METHODS NIH-3T3 mouse embryonic fibroblast cell line was manipulated to knock down or over-express BCAT1 using adenovirus transfection, followed by stimulation with TGFβ1 to induce NIH-3T3 cells transition into myofibroblasts. The expression of fibrosis-related genes was assessed through RT-qPCR, Western blot and immunofluorescence, while fibrosis-related phenotypic changes were evaluated using cell scratch tests, Transwell assays and gel contraction tests.
RESULTS The results showed a significant increase in both BCAT1 mRNA and protein levels upon TGFβ1 stimulation in NIH-3T3 cells (both P<0.01). Knockdown of BCAT1 using shRNA significantly inhibited the expression of TGFβ1-induced fibrosis-related genes Acta2 and Postn mRNA (both P<0.01) as well as fibrosis-related migration and contraction phenotypes (all P<0.01). Conversely, over-expression of BCAT1 increased the expression of these genes and phenotypes (all P<0.01).
CONCLUSION BCAT1 plays a crucial role in regulating the expressions of fibrosis-related genes and phenotypes during the transition of NIH-3T3 fibroblasts towards myofibroblasts, suggesting its potential as a target for fibrosis treatment.
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