PAMM Does not Affect Adipogenic Differentiation of Human Adipose-derived Stem Cells

Objective PAMM(peroxiredoxin-like2activated in M-CSF stimulated monocytes)is a secreted protein which shows high expression in white adipose tissues, but the roles of PAMM in many biological processes are still unknown. To providenew clues for PAMM function research ,this study is intended to investigate the possible role of PAMM in white adipogenesis as well as the downstream genes regulated by PAMM. Methods Adipogenic differentiation and adipogenic inhibition models of human ADSCs(adipose-derived stem cells)were established using adipogenic cocktail(AC)medium or AC plus IL-1 alpha,respectively.Expression of PAMM in ADSCs was suppressed or overexpressed using siRNA interference or plasmid transfection.Gene array,mRNA sequencing and quantitative RT-PCR were employed to detect the mRNA expression level.Western blot was used to evaluateprotein expression and Oil red O staining was adopted to assess lipid droplets accumulation. Results Expression of PAMM was increased following white adipogenic differentiation of ADSCs and decreased following adipogenic inhibition.However,whenPAMM was knocked down or overexpressed before adipogenic differentiation of ADSCs,the downregulation and upregulation ofPAMM expression generally did not exert evident influence on the formation of lipid droplets and the expression of adipogenesis-related genes.Similarly,PAMM knockdown in highly differentiated adipocytes had no obvious effect on cellular morphology and the accumulation of lipid droplets. Finally,a bunch of functional genes and gene sets regulated by PAMM, such asSULF1,A2Mgenesand P53stability gene set,were screened out and confirmed through siRNA interference, mRNA sequencing and qRT-PCR. Conclusion This study suggests PAMM could serve as a useful marker of white adipogenic differentiation of ADSCs,but it exertsno evident effect on white adipogenic differentiation. The unveiled downstream genes and gene sets regulated by PAMM wouldprovide new clues for the functional research of PAMM.