Berberine Chloride and Hyperthermia Promote Osterix Expression and Suppress Cell Cycle Genes in Osteosarcoma Cells

Objective: To investigate the effects of berberine chloride and hyperthermia on the expression of Osterix (Osx), Runt-related transcription factor 2 (RUNX-2), receptor activator of nuclear factor kappa-B ligand (RANKL), cyclin-dependent kinase 2 (CDK2), CDK4, interleukin (IL)-6 and IL-11 genes. Methods: Osteosarcoma cells (MG-63 cells) were treated with three different hyperthermia conditions for 1 h: two groups with mild, two with moderate and two with severe hyperthermia (at 39, 43 and 45 °C, respectively). Berberine chloride (80 μg/ ml) was used for treating one group of mild, moderate and severe hyperthermia (combination). All treated groups were recovered at 37°C for 24 h. Another exposure to hyperthermia (1 h) and recovery at 37°C for 3 h were applied. Gene expression levels were then determined using RT2 PCR profiler arrays. Results: The expression of Osx was highly upregulated in all groups except in the groups treated with severe hyperthermia and mild hyperthermia in combination with berberine chloride. Mild hyperthermia alone induced a slight increase in the expression of RUNX2, whereas severe hyperthermia significantly suppressed RUNX2 levels. Berberine alone significantly induces the up-regulation of RANKL expression. On the other hand, CDK2 mRNA was downregulated in all groups. CDK4 was equally regulated in mild hyperthermia compared to the control group. However, the expression was significantly downregulated in other groups, especially in severe hyperthermia. IL-6 was significantly upregulated in berberine group and all groups of combinations, whereas mild and moderate hyperthermia significantly induced the expression of IL-11 mRNA. Conclusions: Hyperthermia and berberine chloride can promote osteosarcoma cells differentiation and suppresses cell cycle genes.