Abstract
Purpose
In this study, we investigated the correlation between p53 and bcl-2 in gambogic acid (GA)-induced apoptosis.
Method
MTT assay was employed to evaluate MCF-7 cell viability after GA treatment. Cell morphological changes were observed follow-up under the inverted microscope after GA withdrawal. To observe the cell apoptosis, DAPI staining was used. Meanwhile, p53 small interfering RNA (si-RNA) was adopted to knock p53 down. All expressions of p53 and bcl-2 were evaluated by Western blot analysis.
Results
MTT assay showed that GA inhibited MCF-7 cell growth effectively in a time-dependent manner. With 0.5 h GA treatment, p53 was significantly increased, whereas bcl-2 was reduced potently with 6 h GA treatment. After GA withdrawal, p53 expressions were maintained in high levels. Furthermore, bcl-2 decreasing was attenuated after co-treatment with PFT alpha, a p53 transcription blocker. Same result was observed after p53 knock-down by p53 si-RNA.
Conclusions
Gambogic acid induced human breast cancer cells MCF-7 apoptosis by reducing bcl-2 expression via p53.
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Acknowledgments
We thank Professor Zhijie Chang for helpful comments and technical suggestions and Dr. Feng Feng for gambogic acid purification. This work was supported by the Natural Science Foundation of China (No. 30472044), the Natural Science Foundation of Jiangsu Province, China (No. BK2005096), Service Center for Pharmacodynamics Research and Evaluation of Jiangsu Province.
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432_2009_624_MOESM1_ESM.ppt
S1. GA with 4μM is sufficient to induce p53. MCF-7 cancer cells were treated with GA with the concentration of 2, 4, and 8 μM for 6 h and Western blot assay was performed as described in methods and materials. (PPT 60 kb)
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Gu, H., Rao, S., Zhao, J. et al. Gambogic acid reduced bcl-2 expression via p53 in human breast MCF-7 cancer cells. J Cancer Res Clin Oncol 135, 1777–1782 (2009). https://doi.org/10.1007/s00432-009-0624-2
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DOI: https://doi.org/10.1007/s00432-009-0624-2