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Differential proteomic analysis of HeLa cells treated with Honokiol using a quantitative proteomic strategy

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Summary.

Honokiol (HNK) is an active component purified from Magnolia officinalis. HNK exhibits antitumor effects by inducing apoptosis and inhibiting the growth of many cancer cell lines, while proteins involved in antitumor effects in proteomic level are still unclear. In our study, HNK could inhibit HeLa cell proliferation and induce apoptosis in a concentration- and time-dependent manner. We utilized a quantitative proteomic technique termed SILAC (Stable isotope labeling with amino acids in cell culture)-MS (mass spectrometry) to study the differential proteomic profiling of HeLa cells treated by HNK. A total of 85 proteins were changed after HeLa cells were treated with 12 µg/ml HNK for 8 h, and 8 proteins showed up-regulation while 77 proteins down-regulated. The changed proteins were classified into 9 different categories, which covered a broad variety of cellular functions. In conclusion, HNK performs cytotoxicity to HeLa cells through co-operating of many proteins and different pathways.

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Abbreviations

ACN:

acetonitrile

DMSO:

dimethyl sulfoxide

2-D PAGE:

2-dimensional polyacrylamide gel electrophoresis

FCM:

Flow cytometry

HNK:

honokiol

LC-MS/MS:

liquid chromatography followed by tandem mass spectrometry

Leu-d0:

normal leucine

Leu-d3:

deuterated leucine

MALDI-TOF:

matrix assisted laser desorption ion-time of flight

MS:

mass spectrometry

MTT:

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

PBS:

phosphate-buffered-saline

PBST:

phosphate-buffered-saline with 1% Tween

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

14-3-3 sigma:

14-3-3σ

SILAC:

Stable isotope labeling with amino acids in cell culture

TFA:

trifluoroacetic acid

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Authors and Affiliations

  1. West China Second Hospital, and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China

    B. Ling, S.-F. Liang, Y.-H. Xu, X.-Y. Zhao, M.-H. Tang, X.-Y. Liu, X. Zhao, C.-H. Huang, L.-J. Chen & Y.-Q. Wei

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  1. B. Ling
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  2. S.-F. Liang
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  3. Y.-H. Xu
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  4. X.-Y. Zhao
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  5. M.-H. Tang
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  6. X.-Y. Liu
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  7. X. Zhao
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  8. C.-H. Huang
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  9. L.-J. Chen
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  10. Y.-Q. Wei
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Additional information

The first two authors contribute equally to the paper. This article contains electronic supplementary material (see the online version of this paper under www.springerlink.com).

Authors’ address: Xia Zhao, Shufang Liang, West China Second Hospital, and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, 1# Keyuan Road 4, Gaopeng Street, Chengdu 610041, China

Electronic supplementary material

Table 1

. All changed proteins quantified by SILAC-MS

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Ling, B., Liang, SF., Xu, YH. et al. Differential proteomic analysis of HeLa cells treated with Honokiol using a quantitative proteomic strategy. Amino Acids 35, 115–122 (2008). https://doi.org/10.1007/s00726-007-0615-z

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  • DOI: https://doi.org/10.1007/s00726-007-0615-z

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