Abstract
Background
Besides Mad2’s role in carcinogenesis, recent study has shown that it is essential in cell survival. Here we found that knockdown of Mad2 causes osteosarcoma cell death through apoptosis, with the apoptotic signal resulting from Rad21 cleavage.
Methods
U2OS and MG63 cells were divided into three groups: the Mad2 siRNA group, mock group and normal control group; the Mad2 siRNA group and mock group are transfected with Mad2 shRNA plasmid and mock plasmid, respectively. G418 was used to increase the transfection efficacy, which was evaluated by GFP fluorescence. Quantitative PCR and Western blotting analyses were used to detect the transcription and expression of Mad2, Rad21 and caspase-3, respectively. Flow cytometry assay using PE-labeled Annexin-V and PI, TUNEL assay and Hoechst 33258 staining were used to evaluate cell apoptosis.
Results
We successfully achieved knockdown of Mad2 expression in cancer cells using RNA interference. We observed obvious apoptosis in the Mad2 siRNA group compared with the Mock and control group. We found that the apoptosis induced by Mad2 knockdown correlated with Rad21 cleavage.
Conclusion
These results confirmed that knockdown of Mad2 causes osteosarcoma cell death through apoptosis and provides evidence that the apoptotic signal resulted from Rad21 cleavage. This study suggested that Mad2 has potential to be a novel target for cancer therapy.
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Acknowledgments
We thank Dr. Zan Tong (Wuhan University, Wuhan, China) for her critical reading of this manuscript. This work was supported by a grant from the Natural Science Foundation of Hubei Province, China (no. 302-131702).
Conflict of interest
None of the authors of this manuscript have received any type of support, benefits or funding from any commercial party related directly or indirectly to the subject of this article.
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Yu, L., Guo, W., Zhao, S. et al. Knockdown of Mad2 induces osteosarcoma cell apoptosis-involved Rad21 cleavage. J Orthop Sci 16, 814–820 (2011). https://doi.org/10.1007/s00776-011-0156-x
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DOI: https://doi.org/10.1007/s00776-011-0156-x