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hPNAS-4 inhibits proliferation through S phase arrest and apoptosis: underlying action mechanism in ovarian cancer cells

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Abstract

PNAS-4, a novel pro-apoptotic gene, was activated during the early response to DNA damage. Previous studies have shown that hPNAS-4 can inhibit tumor growth when over-expressed in ovarian cancer cells. However, the underlying action mechanism remains elusive. In this work, we found that hPNAS-4 expression was significantly increased in SKOV3 cells when exposed to cisplatin, methyl methanesulfonate or mitomycin C, and that its overexpression could induce proliferation inhibition, S phase arrest and apoptosis in A2780s and SKOV3 ovarian cancer cells. The S phase arrest caused by hPNAS-4 was associated with up-regulation of p21. p21 is p53-dispensable and correlates with activation of ERK, and activation of the Cdc25A-Cdk2-Cyclin E/Cyclin A pathway, while the pro-apoptotic effects of hPNAS-4 were mediated by activation of caspase-9 and -3 other than caspase-8, and accompanied by release of AIF, Smac and cytochrome c into the cytosol. Taken together, these data suggest a new mechanism by which hPNAS-4 inhibits proliferation of ovarian cancer cells by inducing S phase arrest and apoptosis via activation of Cdc25A-Cdk2-Cyclin E/Cyclin A axis and mitochondrial dysfunction-mediated caspase-dependent and -independent apoptotic pathways. To our knowledge, we provide the first molecular evidence for the potential application of hPNAS-4 as a novel target in ovarian cancer gene therapy.

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Abbreviations

hPNAS-4:

Human PNAS-4

pc3.1:

pcDNA3.1 expression plasmid

pc3.1-hPNAS-4:

pcDNA3.1 plasmid encoding Human PNAS-4 gene

ATCC:

American Type Culture Collection

DMEM:

Dulbecco’s modified Eagle’s medium

RPMI 1640:

Roswell Park Memorial Institute 1640 medium

MMS:

Methyl methanesulfonate

MMC:

Mitomycin C

FBS:

Fetal bovine serum

EdU:

5-Ethyl-2′-deoxyuridine

CDK:

Cyclin-dependent kinase

Cyt C:

Cytochrome C

Smac:

Second mitochondria-derived activator of caspase

AIF:

Apoptosis induced factor

Cox-IV:

Cytochrome oxidase subunit IV

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Acknowledgments

The authors thank Dr. Yong-qiu Mao for her technical support. This work was supported by Natural Science Foundation of China (30900744).

Conflict of interest

No potential conflicts of interest were disclosed.

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Authors and Affiliations

  1. State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, 1# Keyuan Road 4, Gaopeng Street, High Technological Development Zone, Chengdu, 610041, China

    Lei Li, Chao Lin, Yang Wan, Xin-yu Zhao, Chun-lai Nie, Zhu Yuan & Yu-quan Wei

  2. Teaching Center of Clinical Experiment, Chengdu Medical College, Chengdu, 610083, China

    Den-bang Chen & Kang Cao

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  1. Lei Li
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  2. Den-bang Chen
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  3. Chao Lin
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  5. Yang Wan
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  7. Chun-lai Nie
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Corresponding author

Correspondence to Zhu Yuan.

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Lei Li and Den-bang Chen contributed equally to this study.

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Li, L., Chen, Db., Lin, C. et al. hPNAS-4 inhibits proliferation through S phase arrest and apoptosis: underlying action mechanism in ovarian cancer cells. Apoptosis 18, 467–479 (2013). https://doi.org/10.1007/s10495-012-0797-z

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