Effects of AEBP1 knockdown on gene expression profiles in human skeletal muscle myoblasts

We recently identified adipocyte enhancer binding protein 1 (AEBP1) as a novel cancer stroma-associated gene and reported that AEBP1 activates cancer-associated fibroblasts in oral cancer. In this study, we show that AEBP1 is negatively associated with skeletal muscle cell differentiation. We found that AEBP1 is downregulated during the diffraction of human skeletal muscle myoblasts (HSMMs). Microarray analysis revealed that knockdown of AEBP1 significantly upregulated a number of genes associated with myogenesis in HSMMs. Notably, co-culture of HSMMs with oral squamous cell carcinoma cells resulted in downregulation of muscle-related genes and upregulation of AEBP1 in HSMMs. For knockdown of AEBP1, 5^10E4 cells were transfected with Silencer Select Pre-designed siRNA (100 pmol each; AEBP1 siRNA1, s1145; AEBP1 siRNA2, s1146; Thermo Fisher Scientific) or Silencer Select Negative Control No. 1 siRNA (Thermo Fisher Scientific) using TransIT-X2 Dynamic Delivery System (Mirus Bio, Madison, WI, USA). Cells were harvested 72 h after transfection. For co-culture experiments, HSMMs (5^10E4 cells/well in 6-well plates) were indirectly co-cultured with OSCC cells (1^10E5 cells in culture inserts) for 72 h.