23 November 2016 : Animal Research
Med Sci Monit 2016; 22:4516-4522
BACKGROUND: Previous studies indicated that calreticulin (CRT) regulated various biological processes. This study was aimed to investigate the function of CRT in Schwann cells (SCs).
MATERIAL AND METHODS: SCs were separated from sciatic nerves of mice and were transfected with pcDNA3.1-CRT (pc-CRT), small interfering RNA targets CRT (siCRT), or their corresponding negative controls. The expression of CRT was determined by quantitative reverse transcription PCR (qRT-PCR) and Western blot analysis. Then, cell proliferation, migration, and apoptosis were measured by 3-(4, 5-dimethylhiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, modified 2-chamber migration assay, and flow cytometry, respectively. Finally, the phosphorylation levels of key kinases in the phosphatidylinositol-3-kinase (PI3K)/AKT and the extracellular signal-regulated kinase/ribosomal S6 kinase 2 (ERK/S6) pathways were detected by Western blot analysis.
RESULTS: Overexpression of CRT remarkably increased viability (P<0.05, P<0.01 or P<0.001) and migration (P<0.001), but inhibited apoptosis (P<0.05). The CRT-knockdown showed the inverse impacts on viability (P<0.05 or P<0.001), migration (P<0.001), and apoptosis (P<0.001). Additionally, the phosphorylation levels of AKT (Thr308 and Ser473), ERK, and S6 were all up-regulated in CRT-overexpressed cells (P<0.001), and were down-regulated in CRT-knockdown cells (P<0.05, P<0.01 or P<0.001).
CONCLUSIONS: Overexpression of CRT in SCs promoted cell proliferation and migration but suppressed cell apoptosis. The PI3K/AKT and ERK/S6 pathways might be involved in the functional effects of CRT on SCs.
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