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Effect of bone sialoprotien on PI3K-AKT signaling pathway of breast cancer MDA-MB-231 cells
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Abstract:
Objective:To investigate the effect of bone sialoprotein (BSP) on the PI3K-AKT signaling pathway in breast cancer MDA-MB-231 cells. Methods: BSP-silenced MDA-MB-231 cells were treated with recombinant human BSP (rhBSP) and the PI3K-AKT specific inhibitor LY294002. Western blotting analysis was used to detect the phosphorylation of AKT, qPCR was conducted to evaluate caspase-3, cyclin D1 mRNA expressions, and the proliferation of cells was analyzed by MTT assay. Results: Compared with the 231BO-Scrambled cells in control group, BSP protein expression in BSP-silenced 231BO-BSP27 cells was significantly lower (74.32±2.18)% (P<0.01), and expression level of AKT phosphorylation was also significantly lower (33.30±2.61) % (P<0.01), resulting in up-regulation of caspase-3 mRNA level (1.000±0.000 vs 1.733±0.039, P<0.01) , down-regulation of cyclin D1 mRNA (1.000±0.000 vs 0370±0.012, P<0. 01),and the inhibition of 231BO-BSP27 cells growth (P<0.05). After treatment with exogenous rhBSP, the phosphorylation of AKT was increased in both 231BO-Scrambled and 231BO-BSP27 cells \[(17.86±2.27)%, (33.78±1.51)%, P<0.01\]. rhBSP treatment decreased caspase-3 mRNA (1.000±0.039 vs 0.541±0.091, P<0.01) , and increased cyclin D1 mRNA (1.000±0.000 vs 2.921±0.032, P<0.01) expression in 231BO-BSP27 cells, and stimulated the proliferation of 231BO-Scrambled and 231BO-BSP27 cells (P<0.01). Furthermore, rhBSP-induced activation of AKT was reversed by LY294002 in 231BO-Scrambled and 231BO-BSP27 cells (P<0.05), with an increase in caspase-3 mRNA and decrease in cyclin D1 mRNA expression in 231BO-BSP27 cells (all P<0.01), causing proliferation inhibition in 231BO-Scrambled and 231BO-BSP27 cells (P<0.01). Conclusion: BSP can regulate the mRNA expressions of caspase-3 and cyclin D1, and affect the proliferation of breast cancer MDA-MB-231 cells through the PI3K-AKT signaling pathway.
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Project Supported:
Project supported by Natural Science Foundation of Guangdong Province (No.06104396)
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