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

Objectives: The aim of the current study was to investigate the effects of berberine chloride and various heat conditions on the gene expression of Osterix, RUNX-2, RANKL, CDK2, CDK4, IL-6 and IL-11. Methods:  Six groups of cells were treated with hyperthermia for 1 h: Two groups at mild, two at moderate and two at severe hyperthermia (39, 43 and 45°C respectively). Berberine chloride (80 µg/mL) was selected for treating one group of mild, moderate and severe hyperthermia (combination). All treated groups were recovered at 37°C for 24 h. Another exposure for hyperthermia (1 h) and recovery for 3 h at 37°C were applied. Results: The expression of Osterix was highly upregulated in all groups except in the severe hyperthermia and mild hyperthermia with berberine groups. Only the mild hyperthermia without berberine induced a slight increase in the expression of RUNX2, whereas severe hyperthermia alone suppressed the levels on a significant manner. Berberine alone was more effective in significantly up-regulating RANKL expression. On the other hand, CDK2 mRNA was downregulated in all groups. CDK4 showed a similar regulation in the mild hyperthermia group with control, but the expression was downregulated in the other groups, especially in severe hyperthermia the expression was significantly downregulated (p<0.5). IL-6 was upregulated highly and significantly in the group of berberine and all groups of combinations, whereas mild and moderate hyperthermia stimulated significant expression of IL-11 mRNA. Conclusion:  These results suggest that hyperthermia and berberine chloride can promote osteosarcoma cells differentiation and arrest cell-cycle

Mild hyperthermia (39°C) attenuates berberine chloride cytotoxicity against osteosarcoma cells

Many studies reported that antioxidants and dietary supplements increase tumour progress and reduce survival. Hyperthermia is an adjuvant treatment to sensitise cancer cells for radio- or chemotherapy. The current study was carried out to determine the effects of berberine chloride and hyperthermia on bone cancer cells. MG-63 osteosarcoma cells were treated with hyperthermia (39, 43 and 45°C, respectively) for 1 h. Then, the cells were treated with a low toxic dose of berberine chloride (80 μg/mL). After that, all treated groups were recovered at 37°C for 24 h. Finally, all groups were treated with hyperthermia (39, 43 and 45°C) for the second time (1 h) and were recovered for 3 h at 37°C. Cells exposed to hyperthermia without treatment of berberine chloride were used as hyperthermia control. Cells treated with 80 μg/mL of berberine at 37°C served as berberine control and cells incubated at 37°C were used as an untreated control. All treated groups showed significant apoptosis compared to the control group (p<0.05) except 39°C. On the other hand, mild hyperthermia treatment (39°C) resulted in a reduction of berberine-induced apoptosis (p<0.001). Severe and moderate hyperthermia did not show a significant increase in the rate of apoptosis compared to berberine treated cells. Mild hyperthermia treatment can effectively reduce berberine cytotoxicity and implying negative effects on cancer therapy