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Proteasome inhibitor MG132 induces thyroid cancer cell apoptosis by modulating the activity of transcription factor FOXO3a

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Abstract

Proteasome inhibitors are promising antitumor drugs with preferable cytotoxicity in malignant cells and have exhibited clinical efficiency in several hematologic malignancies. P53-dependent apoptosis has been reported to be a major mechanism underlying. However, apoptosis can also be found in cancer cells with mutant-type p53, suggesting the involvement of p53-independent mechanism. Tumor suppressor forkhead Box O3 is another substrate of proteasomal degradation, which also functions partially through inducing apoptosis. The aim of this study was to explore the effect of proteasome inhibition on the expression and activity of forkhead Box O3 in thyroid cancer cells. Using flow cytometry, western blot, immunofluorescence staining and quantitative RT-PCR assays, we assessed proteasome inhibitor MG132-induced apoptosis in thyroid cancer cells and its effect on the expression and activity of forkhead Box O3. The resulted showed that MG132 induced significant apoptosis, and caused the accumulation of p53 protein in both p53 wild-type and mutant-type thyroid cancer cell lines, whereas the proapoptotic targets of p53 were transcriptionally upregulated only in the p53 wild-type cells. Strikingly, upon MG132 administration, the accumulation and nuclear translocation of transcription factor forkhead Box O3 as well as transcriptional upregulation of its proapoptotic target genes were found in thyroid cancer cells regardless of p53 status. Cell apoptosis was enhanced by ectopic overexpression while attenuated by silencing of forkhead Box O3. Altogether, we demonstrated that proteasome inhibitor MG132 induces thyroid cancer cell apoptosis at least partially through modulating forkhead Box O3 activity.

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Abbreviations

ATC:

Anaplastic thyroid cancer

DMSO:

Dimethylsulfoxide

FOXO3a:

Forkhead Box O3

GADD45a:

Growth Arrest And DNA-Damage-Inducible 45 Alpha

MAPK:

RAS/RAF/MEK/ERK

MDM2:

Mouse Double Minute 2

PBS:

Phosphate-buffered saline

PDTC:

Poorly-differentiated thyroid cancer

PI3K/Akt:

phosphatidylinositol-3-kinase/Akt

qRT-PCR:

Quantitative RT-PCR

WDTC:

Well-differentiated thyroid cancer

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 81372217 and 81572627), Science and Technology Project of Shaanxi Province (No. 2014K11-01-01-09), the Fundamental Research Funds for the Central Universities (No. xjj2015022) and Fund of First Affiliated Hospital of Xi’an Jiaotong University (No. 2014YK2).

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Authors and Affiliations

  1. Department of Endocrinology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, The People’s Republic of China

    Wei Qiang, Fang Sui, Jingjing Ma, Xinru Li, Xiaojuan Ren, Yuan Shao, Jiazhe Liu, Bingyin Shi & Peng Hou

  2. Department of Endocrinology and Metabolism, The First Affiliated Hospital of China Medical University, Shenyang, 110001, The People’s Republic of China

    Haixia Guan

  3. Key Laboratory for Tumor Precision Medicine of Shaanxi Province, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, The People’s Republic of China

    Bingyin Shi & Peng Hou

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  1. Wei Qiang
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Correspondence to Peng Hou.

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Qiang, W., Sui, F., Ma, J. et al. Proteasome inhibitor MG132 induces thyroid cancer cell apoptosis by modulating the activity of transcription factor FOXO3a. Endocrine 56, 98–108 (2017). https://doi.org/10.1007/s12020-017-1256-y

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