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Apoptosis induced by ardipusilloside III through BAD dephosphorylation and cleavage in human glioblastoma U251MG cells

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Abstract

Ardipusilloside III is a saponin newly isolated from Ardisia pusilla A.DC. Since saponins have exhibited broad anti-cancer and pro-apoptotic activity, we investigated the ability of ardipusilloside III to induce apoptosis in human glioblastoma U251MG cells, as well as the involvement of apoptotic signaling pathways. Ardipusilloside III markedly suppressed proliferation of U251MG cells in a time- and dose-dependent manner (P < 0.05, IC50 = 8.2 μg/ml), but did not affect the growth of primary cultures of human astrocytes. Ardipusilloside III-treated U251MG cells underwent typical apoptotic changes. Exposure to a low dose of ardipusilloside III provoked G2/M-phase cell cycle arrest, which preceded apoptosis characterized by the appearance of cells with sub-G1 DNA content. However, a higher dose of ardipusilloside III induced apoptosis without first causing cell cycle arrest. In addition, ardipusilloside III exposure resulted in time-dependent BAD dephosphorylation and cleavage as well as activation of caspase-8 and caspase-3. Therefore, both the intrinsic pathway of apoptosis, mediated by BAD dephosphorylation and cleavage, and the extrinisic pathway of apoptosis, mediated by caspase-8 and caspase-3 activation, were involved in ardipusilloside III-induced apoptosis. These data suggest that ardipusilloside III is a reliable candidate for chemotherapeutic treatment of human glioblastomas, and should be investigated further.

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Abbreviations

IC50 :

50% inhibitory concentration

BAD:

Bcl-xL/Bcl-2-associated death promoter homolog

DMEM:

Dulbecco’s modified Eagle’s medium

MTT:

3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

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Acknowledgments

The authors would like to thank Juan Li for her excellent technical assistance and Rui-Feng Cao for his helpful discussions. This work was funded by Chinese National Natural Science Foundation, Grant number: 20502035.

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Authors and Affiliations

  1. Department of Neurosurgery, Xijing Institute of Clinical Neuroscience, Xijing Hospital, Fourth Military Medical University, No.127 Changle Western Road, Xi’an, 710032, P.R. China

    Hong Lin, Xiang Zhang, Guang Cheng, Wei Zhang, Hai-Ning Zhen, Jin-Xiang Cheng, Bo-Lin Liu, Wei-Dong Cao, Wen-Peng Dong & Peng Wang

  2. Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Xi’an, P.R. China

    Hai-Feng Tang

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  1. Hong Lin
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Correspondence to Xiang Zhang.

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Hong Lin, Xiang Zhang, Guang Cheng, and Hai-Feng Tang contributed equally to the work.

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Lin, H., Zhang, X., Cheng, G. et al. Apoptosis induced by ardipusilloside III through BAD dephosphorylation and cleavage in human glioblastoma U251MG cells. Apoptosis 13, 247–257 (2008). https://doi.org/10.1007/s10495-007-0170-9

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