Skip to main content

Advertisement

SpringerLink
Log in

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

  • Laboratory Investigation
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. 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

    Article  PubMed  Google Scholar 

  2. 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

    Article  CAS  PubMed  Google Scholar 

  3. 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

    Article  CAS  PubMed  Google Scholar 

  4. 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

    Article  CAS  PubMed  Google Scholar 

  5. 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 272:32360–32369

    Article  CAS  PubMed  Google Scholar 

  6. 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

    Article  CAS  PubMed  Google Scholar 

  7. 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

    Article  CAS  PubMed  Google Scholar 

  8. 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

    Article  CAS  PubMed  Google Scholar 

  9. 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

    Article  CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 25:3419–3423

    CAS  PubMed  Google Scholar 

  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

    Article  CAS  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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 18:981–986

    CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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 6:19132–19147

    Article  PubMed  PubMed Central  Google Scholar 

  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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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

    Article  PubMed  Google Scholar 

  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 8:421–430

    Article  CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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

    CAS  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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

    Article  PubMed  Google Scholar 

  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 25:833–837

    CAS  PubMed  Google Scholar 

  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 7:1320–1324

    CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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

    Google Scholar 

  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

    Article  PubMed  PubMed Central  Google Scholar 

  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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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

    Article  CAS  PubMed  Google Scholar 

  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 21:3629–3633

    Article  PubMed  PubMed Central  Google Scholar 

  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 19:4068–4075

    PubMed  Google Scholar 

Download references

Acknowledgements

The study was supported by the Natural Science Foundation of China (NSFC) (No. 81172180).

Author contributions

XYH, LJ and YYQ conceived and designed the experiments. LJ, RSY and LYJ performed all of the experiments. LJ and YYQ analysed the data. LZG and DB provided material and collected the clinical data. LJ was involved in writing the paper. All of the authors gave final approval for the manuscript to be submitted for publication.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, First Affiliated Hospital of Dalian Medical University, 222 Zhong Shan Road, Dalian, 116011, People’s Republic of China

    Jun Li, Siyang Ren, Zhigang Lian, Bin Dong, Yiqun Yao & Yinghui Xu

  2. Department of Interventional Therapy, First Affiliated Hospital of Dalian Medical University, Dalian, People’s Republic of China

    Yongjian Liu

Authors
  1. Jun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Siyang Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yongjian Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhigang Lian
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bin Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yiqun Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yinghui Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yiqun Yao or Yinghui Xu.

Ethics declarations

Conflict of interest

The authors have no financial or personal conflict of interest to declare.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 19 KB)

Supplementary material 2 (XLS 82 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, J., Ren, S., Liu, Y. et al. Knockdown of NUPR1 inhibits the proliferation of glioblastoma cells via ERK1/2, p38 MAPK and caspase-3. J Neurooncol 132, 15–26 (2017). https://doi.org/10.1007/s11060-016-2337-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-016-2337-0

Keywords

Search

Navigation