Oncogenic WIP1 phosphatase attenuates the DNA damage response and sensitizes p53 mutant Jurkat cells to apoptosis

Eren,Mehtap Kilic; Kartal,Nur Betül; Pi̇levneli̇,Hatice

doi:10.3892/ol.2021.12740

Oncogenic WIP1 phosphatase attenuates the DNA damage response and sensitizes p53 mutant Jurkat cells to apoptosis

Authors:
- Mehtap Kilic Eren
- Nur Betül Kartal
- Hatice Pi̇levneli̇
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Affiliations: Department of Medical Biology, Faculty of Medicine, University of Aydin Adnan Menderes, 09010 Aydin, Turkey, Department of Medical Biochemistry, Institute of Health Sciences, University of Aydin Adnan Menderes, 09010 Aydin, Turkey, Department of Medical Biology, Institute of Health Sciences, University of Aydin Adnan Menderes, 09010 Aydin, Turkey
Published online on: April 19, 2021 https://doi.org/10.3892/ol.2021.12740
Article Number: 479
Copyright: © Eren et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Wild‑type (wt) p53‑induced phosphatase 1 (Wip1), encoded by the protein phosphatase, Mg2+/Mn2+ dependent 1D (PPM1D) gene, is a serine/threonine phosphatase induced upon genotoxic stress in a p53‑dependent manner. Wip1/PPM1D is frequently overexpressed, amplified and mutated in human solid tumors harboring wt p53 and is thus currently recognized as an oncogene. Oncogenic Wip1 dampens cellular stress responses, such as cell cycle checkpoints, apoptosis and senescence, and consequently increases resistance to anticancer therapeutics. Targeting Wip1 has emerged as a therapeutic strategy for tumors harboring wt p53. However, little is known about the efficacy of Wip1‑targeted therapies in tumors lacking p53. The present study aimed to investigate the potential role of oncogenic Wip1 in p53 mutant (mt) Jurkat cells. In the present study, it was demonstrated that p53 mt Jurkat cells exhibited PPM1D/Wip1 gene amplification and expressed relatively high levels of Wip1, as confirmed by gene copy number and RNA expression analysis. In addition, Jurkat cells underwent G2 cell cycle arrest, apoptotic cell death and senescence in response to etoposide and doxorubicin, although the phosphorylation levels of DNA damage response (DDR) elements, including ataxia‑telangiectasia mutated, ataxia‑telangiestasia and Rad3‑related, checkpoint kinase (Chk)1 and Chk2 were significantly low. Accordingly, the targeting of Wip1 phosphatase by RNA interference increased the phosphorylation of DDR elements, but decreased the rate of apoptosis in response to etoposide or doxorubicin in Jurkat cells. The induction of senescence or cell cycle arrest was not affected by the knockdown of Wip1. The results suggest that increased Wip1 expression enhances the apoptotic sensitivity of Jurkat cells in response to chemotherapeutic agents by attenuating DDR signaling. The present study highlights the possible pro‑apoptotic role of Wip1 in a p53 mt T‑cell acute lymphoblastic leukemia cell line. The data suggest the careful consideration of future treatment strategies aiming to manipulate or target Wip1 in human cancers lacking p53.

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1	Bartek J, Bartkova J and Lukas J: DNA damage signalling guards against activated oncogenes and tumour progression. Oncogene. 26:7773–7779. 2007. View Article : Google Scholar : PubMed/NCBI
2	Halazonetis TD, Gorgoulis VG and Bartek J: An oncogene-induced DNA damage model for cancer development. Science. 319:1352–1355. 2008. View Article : Google Scholar : PubMed/NCBI
3	Jackson SP and Bartek J: The DNA-damage response in human biology and disease. Nature. 461:1071–1078. 2009. View Article : Google Scholar : PubMed/NCBI
4	Shimada M and Nakanishi M: Response to DNA damage: Why do we need to focus on protein phosphatases? Front Oncol. 3:82013. View Article : Google Scholar : PubMed/NCBI
5	Wang ZP, Tian Y and Lin J: Role of wild-type p53-induced phosphatase 1 in cancer. Oncol Lett. 14:3893–3898. 2107. View Article : Google Scholar : PubMed/NCBI
6	Fiscella M, Zhang HL, Fan S, Sakaguchi K, Shen S, Mercer EW, Vande Woude GF, O'Connor MP and Appella E: Wip1, a novel human protein phosphatase that is induced in response to ionizing radiation in a p53-dependent manner. Proc Natl Acad Sci USA. 94:6048–6053. 1997. View Article : Google Scholar : PubMed/NCBI
7	Chew J, Biswas S, Shreeram S, Humaidi M, Wong ET, Dhillion MK, Teo H, Hazra A, Fang CC, López-Collazo E, et al: WIP1 phosphatase is a negative regulator of NF-kappaB signalling. Nat Cell Biol. 11:659–666. 2009. View Article : Google Scholar : PubMed/NCBI
8	Le Guezennec X and Bulavin DV: WIP1 phosphatase at the crossroads of cancer and aging. Trends Biochem Sci. 35:109–114. 2010. View Article : Google Scholar : PubMed/NCBI
9	Lowe J, Cha H, Lee MO, Mazur SJ, Appella E and Fornace AJ Jr: Regulation of the Wip1 phosphatase and its effects on the stress response. Front Biosci (Landmark Ed). 17:1480–1498. 2012. View Article : Google Scholar : PubMed/NCBI
10	Zhao M, Zhang H, Zhu G, Liang J, Chen N, Yang Y, Liang X, Cai H and Liu W: Association between overexpression of Wip1 and prognosis of patients with non-small cell lung cancer. Oncol Lett. 11:2365–2370. 2016. View Article : Google Scholar : PubMed/NCBI
11	Goloudina AR, Kochetkova EY, Pospelova TV and Demidov ON: Wip1 phosphatase: Between p53 and MAPK kinases pathways. Oncotarget. 7:31563–31571. 2016. View Article : Google Scholar : PubMed/NCBI
12	Yin S, Wang P, Yang L, Liu Y, Wang Y, Liu M, Qi Z, Meng J, Shi TY, Yang G and Zang R: Wip1 suppresses ovarian cancer metastasis through the ATM/AKT/Snail mediated signaling. Oncotarget. 7:29359–29370. 2016. View Article : Google Scholar : PubMed/NCBI
13	Pechackova S, Burdova K, Benada J, Kleiblova P, Jenikova G and Macurek L: Inhibition of WIP1 phosphatase sensitizes breast cancer cells to genotoxic stress and to MDM2 antagonist nutlin-3. Oncotarget. 7:14458–14475. 2016. View Article : Google Scholar : PubMed/NCBI
14	Macurek L, Benada J, Müllers E, Halim VA, Krejčíková K, Burdová K, Pecháčková S, Hodný Z, Lindqvist A, Medema RH and Bartek J: Downregulation of Wip1 phosphatase modulates the cellular threshold of DNA damage signaling in mitosis. Cell Cycle. 12:251–262. 2013. View Article : Google Scholar : PubMed/NCBI
15	Pecháčková S, Burdová K and Macurek L: WIP1 phosphatase as pharmacological target in cancer therapy. J Mol Med (Berl). 95:589–599. 2017. View Article : Google Scholar
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Eren MK and Tabor V: The role of hypoxia inducible factor-1 alpha in bypassing oncogene-induced senescence. PLoS One. 9:e1010642014. View Article : Google Scholar : PubMed/NCBI
18	Drexler HG, Fombonne S, Matsuo Y, Hu ZB, Hamaguchi H and Uphoff CC: p53 alterations in human leukemia-lymphoma cell lines: In vitro artifact or prerequisite for cell immortalization? Leukemia. 14:198–206. 2000. View Article : Google Scholar : PubMed/NCBI
19	Bulavin DV, Demidov ON, Saito S, Kauraniemi P, Phillips C, Amundson SA, Ambrosino C, Sauter G, Nebreda AR, Anderson CW, et al: Amplification of PPM1D in human tumors abrogates p53 tumor-suppressor activity. Nat Genet. 31:210–215. 2002. View Article : Google Scholar : PubMed/NCBI
20	Martin AC, Facchiano AM, Cuff AL, Hernandez-Boussard T, Olivier M, Hainaut P and Thornton JM: Integrating mutation data and structural analysis of the TP53 tumor-suppressor protein. Human Mutat. 19:149–164. 2002. View Article : Google Scholar : PubMed/NCBI
21	Bond GL, Hu W and Levine AJ: MDM2 is a central node in the p53 pathway: 12 years and counting. Curr Cancer Drug Targets. 5:3–8. 2005. View Article : Google Scholar : PubMed/NCBI
22	Lu X, Ma O, Nguyen TA, Jones SN, Oren M and Donehower LA: The Wip1 Phosphatase acts as a gatekeeper in the p53-Mdm2 autoregulatory loop. Cancer Cell. 12:342–54. 2007. View Article : Google Scholar : PubMed/NCBI
23	Goloudina AR, Tanoue K, Hammann A, Fourmaux E, Le Guezennec X, Bulavin DV, Mazur SJ, Appella E, Garrido C and Demidov ON: Wip1 promotes RUNX2-dependent apoptosis in p53-negative tumors and protects normal tissues during treatment with anticancer agents. Proc Natl Acad Sci USA. 109:E68–E75. 2012. View Article : Google Scholar : PubMed/NCBI
24	Zhu YH, Zhang CW, Lu L, Demidov ON, Sun L, Yang L, Bulavin DV and Xiao ZC: Wip1 regulates the generation of new neural cells in the adult olfactory bulb through p53-dependent cell cycle control. Stem Cells. 27:1433–1442. 2009. View Article : Google Scholar : PubMed/NCBI
25	Xia ZS, Wu D, Zhong W, Lu XJ, Yu T and Chen QK: Wip1 gene silencing enhances the chemosensitivity of human colon cancer cells. Oncol Lett. 14:1875–1883. 2017. View Article : Google Scholar : PubMed/NCBI
26	Burma S, Chen BP, Murphy M, Kurimasa A and Chen DJ: ATM phosphorylates histone H2AX in response to DNA double-strand breaks. J Biol Chem. 276:42462–42467. 2001. View Article : Google Scholar : PubMed/NCBI
27	Mukherjee B, Kessinger C, Kobayashi J, Chen BP, Chen DJ, Chatterjee A and Burma S: DNA-PK phosphorylates histone H2AX during apoptotic DNA fragmentation in mammalian cells. DNA Repair. 5:575–590. 2006. View Article : Google Scholar : PubMed/NCBI
28	Wang H, Wang M, Wang H, Böcker W and Iliakis G: Complex H2AX phosphorylation patterns by multiple kinases including ATM and DNA-PK in human cells exposed to ionizing radiation and treated with kinase inhibitors. J Cell Physiol. 202:492–502. 2005. View Article : Google Scholar : PubMed/NCBI
29	Pilié PG, Tang C, Mills GB and Yap TA: State-of-the-art strategies for targeting the DNA damage response in cancer. Nat Rev Clin Oncol. 16:81–104. 2019. View Article : Google Scholar
30	Kilic M and Schmitt CA: Exploiting drug induced senescence in transgenic mouse models. Beyond Apoptosis: Cellular Outcomes of Cancer Therapy. Roninson IB, Brown JM and Bredesen DE: Informa Health Care Publication; pp. 273–295. 2008, View Article : Google Scholar
31	Crescenzi E, Raia Z, Pacifico F, Mellone S, Moscato F, Palumbo G and Leonardi A: Down-regulation of wild-type p53-induced phosphatase 1 (Wip1) plays a critical role in regulating several p53-dependent functions in premature senescent tumor cells. J Biol Chem. 288:16212–24. 2013. View Article : Google Scholar : PubMed/NCBI