Research Paper Volume 12, Issue 22 pp 23114—23128

Intracellular and extracellular S100A9 trigger epithelial-mesenchymal transition and promote the invasive phenotype of pituitary adenoma through activation of AKT1

Figure 3. Intracellular S100A9 regulates phosphorylation of AKT1Thr308 and induces EMT in PA. (A) The mRNA levels of S100A9 in control group (group 1), NC1 group (group 2), LV-S100A9 group (group 3) were detected by Real-Time PCR(n=9, P***<0.001 versus group 1). (B) Western blot was used to observe S100A9 protein levels in groups 1, 2, and 3(n=5, P***<0.001 versus group 1). (C) The expression of phospho-AKT1Thr308, Vimentin, ICAM-1 and E-cadherin was explored using western blot analysis in groups 1, 2, 3 and a LV-S100A9 + addition of A-674563 group (group 4)(n=5, P***<0.001 vs. group 1. P##<0.01; P###<0.001 vs. group 4). (D, E) Real-time PCR and western blot analysis were used to investigate S100A9 mRNA and protein expression in the control group (group a), NC2 group (group b) and S100A9-shRNA-LV group (group c), respectively(n=5, P***<0.001, compared with group a). (F) Western blot demonstrated that downregulation of S100A9 reduced phospho-AKT1Thr308, Vimentin, ICAM-1, coupled with increase in E-cadherin. The S100A9-shRNA-LV + SC79(AKT1 agonist) group (group d) displayed that the reduced phospho-AKT1Thr308, Vimentin, ICAM-1 and elevated E-cadherin were reversed partially by SC97(n=5. P***<0.001, compared with group a. P##<0.01; P###<0.001, compared with group d).