Top

Published in:

01-03-2017 | Laboratory Investigation

Knockdown of NUPR1 inhibits the proliferation of glioblastoma cells via ERK1/2, p38 MAPK and caspase-3

Authors: Jun Li, Siyang Ren, Yongjian Liu, Zhigang Lian, Bin Dong, Yiqun Yao, Yinghui Xu

Published in: Journal of Neuro-Oncology | Issue 1/2017

Abstract

Nuclear protein-1 (NUPR1), located on chromosome 16p11.2, is a stress response factor that plays an important role in the growth and migration of human malignant tumor cells. However, the role of NUPR1 in glioblastoma remains poorly understood. The expression level of NUPR1 was detected by quantitative real-time PCR and immunohistochemistry (IHC). Wound healing, MTT, cell counting and BrdU assays were used to analyze the migration and proliferation of glioblastoma cells after down-regulating NUPR1 expression using a lentiviral vector. FACS analysis and a signaling antibody array kit were used to detect the mechanism by which NUPR1 modulates cell cycle and apoptosis activities in glioblastoma cells. We confirmed that NUPR1 was up-regulated in glioblastoma tissues compared to NB tissues. Down-regulation of NUPR1 suppressed cell migration and proliferation, arrested the cell cycle in the G0/G1 phase and promoted apoptosis in U251 and U87 cells in vitro. Furthermore, the expression levels of phosphorylated ERK1/2, p38 MAPK and cleaved caspase-3 were decreased upon silencing NUPR1 expression in U251 and U87 cells. In summary, NUPR1 plays an important role in the growth and migration of human glioblastoma cells. Knockdown of NUPR1 suppressed glioblastoma cell growth by arresting the cell cycle and inducing cell apoptosis via decreases in the expression of ERK1/2, p38 MAPK and caspase-3.

Available only for authorised users

Sathornsumetee S, Rich JN, Reardon DA (2007) Diagnosis and treatment of high-grade astrocytoma. Neurol Clin 25:1111–1139. doi:10.1016/j.ncl.2007.07.004 CrossRefPubMed

Lefranc F, Sadeghi N, Camby I, Metens T, Dewitte O, Kiss R (2006) Present and potential future issues in glioblastoma treatment. Expert Rev Anticancer Ther 6:719–732. doi:10.1586/14737140.6.5.719 CrossRefPubMed

Ohgaki H, Dessen P, Jourde B, Horstmann S, Nishikawa T, Di Patre PL, Burkhard C, Schuler D, Probst-Hensch NM, Maiorka PC, Baeza N, Pisani P, Yonekawa Y, Yasargil MG, Lutolf UM, Kleihues P (2004) Genetic pathways to glioblastoma: a population-based study. Cancer Res 64:6892–6899. doi:10.1158/0008-5472.can-04-1337 CrossRefPubMed

Jiang T, Mao Y, Ma W, Mao Q, You Y, Yang X, Jiang C, Kang C, Li X, Chen L, Qiu X, Wang W, Li W, Yao Y, Li S, Li S, Wu A, Sai K, Bai H, Li G, Chen B, Yao K, Wei X, Liu X, Zhang Z, Dai Y, Lv S, Wang L, Lin Z, Dong J, Xu G, Ma X, Cai J, Zhang W, Wang H, Chen L, Zhang C, Yang P, Yan W, Liu Z, Hu H, Chen J, Liu Y, Yang Y, Wang Z, Wang Z, Wang Y, You G, Han L, Bao Z, Liu Y, Wang Y, Fan X, Liu S, Liu X, Wang Y, Wang Q (2016) CGCG clinical practice guidelines for the management of adult diffuse gliomas. Cancer Lett 375:263–273. doi:10.1016/j.canlet.2016.01.024 CrossRefPubMed

Mallo GV, Fiedler F, Calvo EL, Ortiz EM, Vasseur S, Keim V, Morisset J, Iovanna JL (1997) Cloning and expression of the rat p8 cDNA, a new gene activated in pancreas during the acute phase of pancreatitis, pancreatic development, and regeneration, and which promotes cellular growth. J Biol Chem 272:32360–32369CrossRefPubMed

Cano CE, Hamidi T, Sandi MJ, Iovanna JL (2011) Nupr1: the Swiss-knife of cancer. J Cell Physiol 226:1439–1443. doi:10.1002/jcp.22324 CrossRefPubMed

Encinar JA, Mallo GV, Mizyrycki C, Giono L, Gonzalez-Ros JM, Rico M, Canepa E, Moreno S, Neira JL, Iovanna JL (2001) Human p8 is a HMG-I/Y-like protein with DNA binding activity enhanced by phosphorylation. J Biol Chem 276:2742–2751. doi:10.1074/jbc.M008594200 CrossRefPubMed

Garcia-Montero A, Vasseur S, Mallo GV, Soubeyran P, Dagorn JC, Iovanna JL (2001) Expression of the stress-induced p8 mRNA is transiently activated after culture medium change. Eur J Cell Biol 80:720–725. doi:10.1078/0171-9335-00209 CrossRefPubMed

Hoffmeister A, Ropolo A, Vasseur S, Mallo GV, Bodeker H, Ritz-Laser B, Dressler GR, Vaccaro MI, Dagorn JC, Moreno S, Iovanna JL (2002) The HMG-I/Y-related protein p8 binds to p300 and Pax2 trans-activation domain-interacting protein to regulate the trans-activation activity of the Pax2A and Pax2B transcription factors on the glucagon gene promoter. J Biol Chem 277:22314–22319. doi:10.1074/jbc.M201657200 CrossRefPubMed

10.

Goruppi S, Patten RD, Force T, Kyriakis JM (2007) Helix-loop-helix protein p8, a transcriptional regulator required for cardiomyocyte hypertrophy and cardiac fibroblast matrix metalloprotease induction. Mol Cell Biol 27:993–1006. doi:10.1128/mcb.00996-06 CrossRefPubMed

11.

Jung SH, Lee A, Yim SH, Hu HJ, Choe C, Chung YJ (2012) Simultaneous copy number gains of NUPR1 and ERBB2 predicting poor prognosis in early-stage breast cancer. BMC Cancer 12:382. doi:10.1186/1471-2407-12-382 CrossRefPubMedPubMedCentral

12.

Ito Y, Yoshida H, Motoo Y, Iovanna JL, Tomoda C, Uruno T, Takamura Y, Miya A, Kobayashi K, Matsuzuka F, Kuma K, Miyauchi A (2005) Expression of p8 protein in medullary thyroid carcinoma. Anticancer Res 25:3419–3423PubMed

13.

Guo X, Wang W, Hu J, Feng K, Pan Y, Zhang L, Feng Y (2012) Lentivirus-mediated RNAi knockdown of NUPR1 inhibits human nonsmall cell lung cancer growth in vitro and in vivo. Anat Rec 295:2114–2121. doi:10.1002/ar.22571 CrossRef

14.

Lee YK, Jee BA, Kwon SM, Yoon YS, Xu WG, Wang HJ, Wang XW, Thorgeirsson SS, Lee JS, Woo HG, Yoon G (2015) Identification of a mitochondrial defect gene signature reveals NUPR1 as a key regulator of liver cancer progression. Hepatology 62:1174–1189. doi:10.1002/hep.27976 CrossRefPubMed

15.

Cano CE, Hamidi T, Garcia MN, Grasso D, Loncle C, Garcia S, Calvo E, Lomberk G, Dusetti N, Bartholin L, Urrutia R, Iovanna JL (2014) Genetic inactivation of Nupr1 acts as a dominant suppressor event in a two-hit model of pancreatic carcinogenesis. Gut 63:984–995. doi:10.1136/gutjnl-2013-305221 CrossRefPubMed

16.

Jiang WG, Davies G, Martin TA, Kynaston H, Mason MD, Fodstad O (2006) Com-1/p8 acts as a putative tumour suppressor in prostate cancer. Int J Mol Med 18:981–986PubMed

17.

Veerla S, Panagopoulos I, Jin Y, Lindgren D, Hoglund M (2008) Promoter analysis of epigenetically controlled genes in bladder cancer. Genes Chromosomes Cancer 47:368–378. doi:10.1002/gcc.20542 CrossRefPubMed

18.

Pedrola N, Devis L, Llaurado M, Campoy I, Martinez-Garcia E, Garcia M, Muinelo-Romay L, Alonso-Alconada L, Abal M, Alameda F, Mancebo G, Carreras R, Castellvi J, Cabrera S, Gil-Moreno A, Matias-Guiu X, Iovanna JL, Colas E, Reventos J, Ruiz A (2015) Nidogen 1 and nuclear protein 1: novel targets of ETV5 transcription factor involved in endometrial cancer invasion. Clin Exp Metastasis 32:467–478. doi:10.1007/s10585-015-9720-7 CrossRefPubMed

19.

Di Martino MT, Guzzi PH, Caracciolo D, Agnelli L, Neri A, Walker BA, Morgan GJ, Cannataro M, Tassone P, Tagliaferri P (2015) Integrated analysis of microRNAs, transcription factors and target genes expression discloses a specific molecular architecture of hyperdiploid multiple myeloma. Oncotarget 6:19132–19147CrossRefPubMedPubMedCentral

20.

Hu W, Yang Y, Xi S, Sai K, Su D, Zhang X, Lin S, Zeng J (2015) Expression of CPEB4 in human glioma and its correlations with prognosis. Medicine 94:e979. doi:10.1097/md.0000000000000979 CrossRefPubMedPubMedCentral

21.

Liu Y, Lear T, Iannone O, Shiva S, Corey C, Rajbhandari S, Jerome J, Chen BB, Mallampalli RK (2015) The Proapoptotic F-box protein Fbxl7 regulates mitochondrial function by mediating the ubiquitylation and proteasomal degradation of survivin. J Biol Chem 290:11843–11852. doi:10.1074/jbc.M114.629931 CrossRefPubMedPubMedCentral

22.

Brat DJ, Scheithauer BW, Fuller GN, Tihan T (2007) Newly codified glial neoplasms of the 2007 WHO classification of tumours of the central nervous system: angiocentric glioma, pilomyxoid astrocytoma and pituicytoma. Brain Pathol 17:319–324. doi:10.1111/j.1750-3639.2007.00082.x CrossRefPubMed

23.

Clark DW, Mitra A, Fillmore RA, Jiang WG, Samant RS, Fodstad O, Shevde LA (2008) NUPR1 interacts with p53, transcriptionally regulates p21 and rescues breast epithelial cells from doxorubicin-induced genotoxic stress. Curr Cancer Drug Targets 8:421–430CrossRefPubMed

24.

Carracedo A, Egia A, Guzman M, Velasco G (2006) p8 Upregulation sensitizes astrocytes to oxidative stress. FEBS Lett 580: 1571–1575. doi:10.1016/j.febslet.2006.01.084 CrossRefPubMed

25.

Lee YK, Woo HG, Yoon G (2015) Mitochondrial defect-responsive gene signature in liver-cancer progression. BMB Rep 48:597–598CrossRefPubMedPubMedCentral

26.

Huang H, Zhang Y, Wang X, Zhou X, Li S, Wang M, Ren J (2015) Nuclear protein 1 knockdown inhibits proliferation and migration of HepG2 cells. Chin J Cell Mol Immunol 31:782–786

27.

Goruppi S, Iovanna JL (2010) Stress-inducible protein p8 is involved in several physiological and pathological processes. J Biol Chem 285:1577–1581. doi:10.1074/jbc.R109.080887 CrossRefPubMed

28.

Lorente M, Carracedo A, Torres S, Natali F, Egia A, Hernandez-Tiedra S, Salazar M, Blazquez C, Guzman M, Velasco G (2009) Amphiregulin is a factor for resistance of glioma cells to cannabinoid-induced apoptosis. Glia 57:1374–1385. doi:10.1002/glia.20856 CrossRefPubMed

29.

Ito Y, Yoshida H, Motoo Y, Iovanna JL, Nakamura Y, Kakudo K, Uruno T, Takamura Y, Miya A, Noguchi S, Kuma K, Miyauchi A (2005) Expression of p8 protein in breast carcinoma; an inverse relationship with apoptosis. Anticancer Res 25:833–837PubMed

30.

Su SB, Motoo Y, Iovanna JL, Berthezene P, Xie MJ, Mouri H, Ohtsubo K, Matsubara F, Sawabu N (2001) Overexpression of p8 is inversely correlated with apoptosis in pancreatic cancer. Clin Cancer Res 7:1320–1324PubMed

31.

Malicet C, Giroux V, Vasseur S, Dagorn JC, Neira JL, Iovanna JL (2006) Regulation of apoptosis by the p8/prothymosin alpha complex. Proc Natl Acad Sci USA 103:2671–2676. doi:10.1073/pnas.0508955103 CrossRefPubMedPubMedCentral

32.

Bak Y, Shin HJ, Bak IS, Yoon DY, Yu DY (2015) Hepatitis B virus X promotes hepatocellular carcinoma development via nuclear protein 1 pathway. Biochem Biophys Res Commun 466:676–681. doi:10.1016/j.bbrc.2015.09.082 CrossRefPubMed

33.

Cui Y, Lu P, Song G, Liu Q, Zhu D, Liu X (2016) Involvement of PI3K/Akt, ERK and p38 signaling pathways in emodin-mediated extrinsic and intrinsic human hepatoblastoma cell apoptosis. Food Chem Toxicol 92:26–37. doi:10.1016/j.fct.2016.03.013 CrossRefPubMed

34.

Xu L, Wang L, Wen Z, Wu L, Jiang Y, Yang L, Xiao L, Xie Y, Ma M, Zhu W, Ye R, Liu X (2016) Caveolin-1 is a checkpoint regulator in hypoxia-induced astrocyte apoptosis via Ras/Raf/ERK pathway. Am J Physiol Cell Physiol. doi:10.1152/ajpcell.00309.2015

35.

Zeng Z, Leng T, Feng X, Sun H, Inoue K, Zhu L, Xiong ZG (2015) Silencing TRPM7 in mouse cortical astrocytes impairs cell proliferation and migration via ERK and JNK signaling pathways. PLoS One 10:e0119912. doi:10.1371/journal.pone.0119912 CrossRefPubMedPubMedCentral

36.

Liu Y, Wang F, Liu Y, Yao Y, Lv X, Dong B, Li J, Ren S, Yao Y, Xu Y (2016) RNF135, RING finger protein, promotes the proliferation of human glioblastoma cells in vivo and in vitro via the ERK pathway. Sci Rep 6:20642. doi:10.1038/srep20642 CrossRefPubMedPubMedCentral

37.

Dolka I, Krol M, Sapierzynski R (2016) Evaluation of apoptosis-associated protein (Bcl-2, Bax, cleaved caspase-3 and p53) expression in canine mammary tumors: an immunohistochemical and prognostic study. Res Vet Sci 105:124–133. doi:10.1016/j.rvsc.2016.02.004 CrossRefPubMed

38.

Zhang L, Liang P, Zhang R (2015) Impact of mitochondria-mediated apoptosis in U251 cell cycle arrest in G1 stage and caspase activation. Med Sci Monit 21:3629–3633CrossRefPubMedPubMedCentral

39.

Liu XR, Li YQ, Hua C, Li SJ, Zhao G, Song HM, Yu MX, Huang Q (2015) Oxidative stress inhibits growth and induces apoptotic cell death in human U251 glioma cells via the caspase-3-dependent pathway. Eur Rev Med Pharmacol Sci 19:4068–4075PubMed

Title: Knockdown of NUPR1 inhibits the proliferation of glioblastoma cells via ERK1/2, p38 MAPK and caspase-3
Authors: Jun Li
Siyang Ren
Yongjian Liu
Zhigang Lian
Bin Dong
Yiqun Yao
Yinghui Xu
Publication date: 01-03-2017
Publisher: Springer US
Published in: Journal of Neuro-Oncology / Issue 1/2017
Print ISSN: 0167-594X
Electronic ISSN: 1573-7373
DOI: https://doi.org/10.1007/s11060-016-2337-0

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Knockdown of NUPR1 inhibits the proliferation of glioblastoma cells via ERK1/2, p38 MAPK and caspase-3

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2017

Patterns of care and treatment outcomes of patients with Craniopharyngioma in the national cancer database

Hypofractionated stereotactic radiosurgery for pituitary metastases

Effects of the functional HOTAIR rs920778 and rs12826786 genetic variants in glioma susceptibility and patient prognosis

Development of the pediatric quality of life inventory neurofibromatosis type 1 module items for children, adolescents and young adults: qualitative methods

Survival in pediatric medulloblastoma: a population-based observational study to improve prognostication

Development of a questionnaire measuring instrumental activities of daily living (IADL) in patients with brain tumors: a pilot study