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Medical Oncology

, 37:6 | Cite as

Association and clinicopathologic significance of p38MAPK-ERK-JNK-CDC25C with polyploid giant cancer cell formation

  • Kai Liu
  • Rui Lu
  • Qi Zhao
  • Jiaxing Du
  • Yuwei Li
  • Minying Zheng
  • Shiwu ZhangEmail author
Original Paper

Abstract

Background

We previously showed that cobalt chloride (CoCl2) induction of polyploid giant cancer cells (PGCCs) was characterized by abnormal cell cycle-related protein expression and G2/M arrest. The role of the p38MAPK-ERK-JNK signaling pathway in cell cycle regulation has been reported, but the mechanism by which p38MAPK-ERK-JNK regulates PGCCs formation remains unclear. This study examined p38MAPK-ERK-JNK-CDC25C expression in PGCCs and their daughter and control cells and assessed the clinicopathological significance of p38MAPK, ERK, JNK, and CDC25C expression in human ovarian and breast cancers.

Methods

CoCl2 was used to induce the formation of PGCCs in HEY and BT-549 cells. Western blotting and immunocytochemical staining were used to compare the expression and subcellular localization of p38MAPK, ERK, JNK, and CDC25C in the control group and CDC25C knockdown before and after CoCl2 treatment. The specific combination of p38MAPK and ERK with pCDC25C-Ser216 was detected by immunoprecipitation. In addition, p38MAPK, ERK, JNK, and CDC25C immunohistochemical staining were performed to compare the clinicopathologic significances in 81 cases of ovarian cancer tissue, including 20 cases of primary breast cancer with lymph node metastasis (group I), and their corresponding metastatic lymph nodes (group II), 31 cases of primary breast cancer without metastasis (group III), and 10 cases of benign breast tumors (group IV). Breast tumor tissue from 229 was divided into two groups: 167 cases of primary invasive breast cancer (group 1) and 62 cases of lymph node metastatic breast cancer (group 2).

Results

Compared to the control cells, p38MAPK and JNK expression were higher and CDC25C expression was lower in CoCl2-treated cells. Moreover, ERK displayed a trend of increased expression in HEY PGCCs and decreased expression in BT-549 PGCCs. p38MAPK and ERK regulated CDC25C by phosphorylating the CDC25C-Ser216 site and participated in the G2/M phase transition. Immunohistochemical (IHC) analysis of the ovarian tumor tissues showed significant positive staining rates of p38MAPK (P = 0.001), ERK (P = 0.002), JNK (P = 0.000), and CDC25C (P = 0.000) among the four groups. In breast tumor tissues, the overall expression in p38MAPK (P = 0.029), ERK (P = 0.002), JNK (P = 0.013), and CDC25C (P = 0.001) also differed significantly between the two groups.

Conclusion

The p38MAPK-ERK-JNK signaling pathway was involved in cell cycle progression and the formation of PGCCs by regulation of CDC25C.

Keywords

Polyploid giant cancer cells Cell cycle regulation G2/M arrest CDC25C MAPK 

Abbreviations

PGCCs

Polyploid giant cancer cells

CoCl2

Cobalt chloride

EMT

Epithelial-mesenchymal transition

CSC

Cancer stem cells

FBS

Fetal bovine serum

PBS

Phosphate-buffered saline

ICC

Immunocytochemistry

WB

Western blot

IHC

Immunohistochemistry

ATCC

American Type Culture Collection

PMSF

Phenylmethanesulfonylfluoride fluoride

PI

Protease inhibitor cocktail

CDC25

Cell division cycle 25

MAPK

Mitogen-activated protein kinase

JNK

Mitogen-activated protein kinase 8

Ser

Serine

Notes

Author Contributions

SZ designed the study and contributed to manuscript writing; KL and QZ conducted the experiments and drafted the manuscript. RL participated in the sample collection, statistical analysis and contributed to manuscript writing. DJ helped in the finished of the study. YL and MZ participated in design and coordination and helped to draft the manuscript. All authors have read and approved the final manuscript.

Funding

This work was supported in part by grants from the National Natural Science Foundation of China (81672426), and the foundation of committee on science and technology of Tianjin (17ZXMFSY00120 and 17YFZCSY00700).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

12032_2019_1330_MOESM1_ESM.docx (13 kb)
Supplementary material 1 (DOCX 13 kb)

References

  1. 1.
    Zhang S, Mercado-Uribe I, Xing Z, Sun B, Kuang J, Liu J. Generation of cancer stem-like cells through the formation of polyploid giant cancer cells. Oncogene. 2014;33:116–28.  https://doi.org/10.1038/onc.2013.96.CrossRefPubMedGoogle Scholar
  2. 2.
    Fei F, Li C, Wang X, Du J, Liu K, Li B, Yao P, Li Y, Zhang S. Syncytin 1, CD9, and CD47 regulating cell fusion to form PGCCs associated with cAMP/PKA and JNK signaling pathway. Cancer Med. 2019;8:3047–58.  https://doi.org/10.1002/cam4.2173.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Zhang S, Zhang D, Yang Z, Zhang X. Tumor budding, micropapillary pattern, and polyploidy giant cancer cells in colorectal cancer: current status and future prospects. Stem Cells Int. 2016;2016:4810734.  https://doi.org/10.1155/2016/4810734.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Fei F, Zhang D, Yang Z, Wang S, Wang X, Wu Z, Wu Q, Zhang S. The number of polyploid giant cancer cells and epithelial-mesenchymal transition-related proteins are associated with invasion and metastasis in human breast cancer. J Exp Clin Cancer Res. 2015;34:158.  https://doi.org/10.1186/s13046-015-0277-8.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Fei F, Qu J, Liu K, Li C, Wang X, Li Y, Zhang S. The subcellular location of cyclin B1 and CDC25 associated with the formation of polyploid giant cancer cells and their clinicopathological significance. Lab Invest. 2019;99:483–98.  https://doi.org/10.1038/s41374-018-0157-x.CrossRefPubMedGoogle Scholar
  6. 6.
    Wenzel ES, Singh ATK. Cell-cycle checkpoints and aneuploidy on the path to cancer. In vivo (Athens, Greece). 2018;32:1–5.  https://doi.org/10.21873/invivo.11197.CrossRefGoogle Scholar
  7. 7.
    Chen J. The cell-cycle arrest and apoptotic functions of p53 in tumor initiation and progression. Cold Spring Harbor Perspect Med. 2016;6:a026104.  https://doi.org/10.1101/cshperspect.a026104.CrossRefGoogle Scholar
  8. 8.
    Boutros R, Lobjois V, Ducommun B. CDC25 phosphatases in cancer cells: key players? Good targets? Nat Rev Cancer. 2007;7:495–507.  https://doi.org/10.1038/nrc2169.CrossRefPubMedGoogle Scholar
  9. 9.
    Roux PP, Blenis J. ERK and p38 MAPK-activated protein kinases: a family of protein kinases with diverse biological functions. Microbiol Mol Biol Rev: MMBR. 2004;68:320–44.  https://doi.org/10.1128/mmbr.68.2.320-344.2004.CrossRefPubMedGoogle Scholar
  10. 10.
    Gkouveris I, Nikitakis NG. Role of JNK signaling in oral cancer: a mini review. Tumour Biol. 2017;39:1010428317711659.  https://doi.org/10.1177/1010428317711659.CrossRefPubMedGoogle Scholar
  11. 11.
    Eymin B, Claverie P, Salon C, Brambilla C, Brambilla E, Gazzeri S. p14ARF triggers G2 arrest through ERK-mediated Cdc25C phosphorylation, ubiquitination and proteasomal degradation. Cell Cycle. 2006;5:759–65.  https://doi.org/10.4161/cc.5.7.2625.CrossRefPubMedGoogle Scholar
  12. 12.
    Johnson MD, Reeder JE, O’Connell M. p38MAPK activation and DUSP10 expression in meningiomas. J Clin Neurosci. 2016;30:110–4.  https://doi.org/10.1016/j.jocn.2015.12.031.CrossRefPubMedGoogle Scholar
  13. 13.
    Wang Y, Liu J, Cui J, Xing L, Wang J, Yan X, Zhang X. ERK and p38 MAPK signaling pathways are involved in ochratoxin A-induced G2 phase arrest in human gastric epithelium cells. Toxicol Lett. 2012;209:186–92.  https://doi.org/10.1016/j.toxlet.2011.12.011.CrossRefPubMedGoogle Scholar
  14. 14.
    Lv H, Shi Y, Zhang L, Zhang D, Liu G, Yang Z, Li Y, Fei F, Zhang S. Polyploid giant cancer cells with budding and the expression of cyclin E, S-phase kinase-associated protein 2, stathmin associated with the grading and metastasis in serous ovarian tumor. BMC Cancer. 2014;14:576.  https://doi.org/10.1186/1471-2407-14-576.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Niu N, Zhang J, Zhang N, Mercado-Uribe I, Tao F, Han Z, Pathak S, Multani AS, Kuang J, Yao J, et al. Linking genomic reorganization to tumor initiation via the giant cell cycle. Oncogenesis. 2016;5:e281.  https://doi.org/10.1038/oncsis.2016.75.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Duncan AW, Taylor MH, Hickey RD, Hanlon Newell AE, Lenzi ML, Olson SB, Finegold MJ, Grompe M. The ploidy conveyor of mature hepatocytes as a source of genetic variation. Nature. 2010;467:707–10.  https://doi.org/10.1038/nature09414.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Orr-Weaver TL. When bigger is better: the role of polyploidy in organogenesis. Trends Genet: TIG. 2015;31:307–15.  https://doi.org/10.1016/j.tig.2015.03.011.CrossRefPubMedGoogle Scholar
  18. 18.
    Geigl JB, Obenauf AC, Schwarzbraun T, Speicher MR. Defining ‘chromosomal instability’. Trends Genet: TIG. 2008;24:64–9.  https://doi.org/10.1016/j.tig.2007.11.006.CrossRefPubMedGoogle Scholar
  19. 19.
    Holland AJ, Cleveland DW. Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis. Nat Rev Mol Cell Biol. 2009;10:478–87.  https://doi.org/10.1038/nrm2718.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Malumbres M, Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009;9:153–66.  https://doi.org/10.1038/nrc2602.CrossRefPubMedGoogle Scholar
  21. 21.
    Nair JJ, van Staden J. Cell cycle modulatory effects of Amaryllidaceae alkaloids. Life Sci. 2018;213:94–101.  https://doi.org/10.1016/j.lfs.2018.08.073.CrossRefPubMedGoogle Scholar
  22. 22.
    Smith J, Tho LM, Xu N, Gillespie DA. The ATM-Chk2 and ATR-Chk1 pathways in DNA damage signaling and cancer. Adv Cancer Res. 2010;108:73–112.  https://doi.org/10.1016/b978-0-12-380888-2.00003-0.CrossRefPubMedGoogle Scholar
  23. 23.
    Sancar A, Lindsey-Boltz LA, Unsal-Kacmaz K, Linn S. Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem. 2004;73:39–85.  https://doi.org/10.1146/annurev.biochem.73.011303.073723.CrossRefPubMedGoogle Scholar
  24. 24.
    Sun Y, Liu WZ, Liu T, Feng X, Yang N, Zhou HF. Signaling pathway of MAPK/ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct Res. 2015;35:600–4.  https://doi.org/10.3109/10799893.2015.1030412.CrossRefPubMedGoogle Scholar
  25. 25.
    Brenner AK, Reikvam H, Lavecchia A, Bruserud O. Therapeutic targeting the cell division cycle 25 (CDC25) phosphatases in human acute myeloid leukemia–the possibility to target several kinases through inhibition of the various CDC25 isoforms. Molecules (Basel, Switzerland). 2014;19:18414–47.  https://doi.org/10.3390/molecules191118414.CrossRefGoogle Scholar
  26. 26.
    Boutros R, Dozier C, Ducommun B. The when and wheres of CDC25 phosphatases. Curr Opin Cell Biol. 2006;18:185–91.  https://doi.org/10.1016/j.ceb.2006.02.003.CrossRefPubMedGoogle Scholar
  27. 27.
    Sur S, Agrawal DK. Phosphatases and kinases regulating CDC25 activity in the cell cycle: clinical implications of CDC25 overexpression and potential treatment strategies. Mol Cell Biochem. 2016;416:33–46.  https://doi.org/10.1007/s11010-016-2693-2.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Palanivel K, Kanimozhi V, Kadalmani B. Verrucarin A alters cell-cycle regulatory proteins and induces apoptosis through reactive oxygen species-dependent p38MAPK activation in the human breast cancer cell line MCF-7. Tumour Biol. 2014;35:10159–67.  https://doi.org/10.1007/s13277-014-2286-1.CrossRefPubMedGoogle Scholar
  29. 29.
    Zeke A, Misheva M, Remenyi A, Bogoyevitch MA. JNK signaling: regulation and functions based on complex protein-protein partnerships. Microbiol Mol Biol Rev: MMBR. 2016;80:793–835.  https://doi.org/10.1128/mmbr.00043-14.CrossRefPubMedGoogle Scholar
  30. 30.
    Zhou YY, Li Y, Jiang WQ, Zhou LF. MAPK/JNK signalling: a potential autophagy regulation pathway. Biosci Rep. 2015.  https://doi.org/10.1042/bsr20140141.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Bubici C, Papa S. JNK signalling in cancer: in need of new, smarter therapeutic targets. Br J Pharmacol. 2014;171:24–37.  https://doi.org/10.1111/bph.12432.CrossRefPubMedGoogle Scholar
  32. 32.
    Yang Z, Cheng B, Song J, Wan Y, Wang Q, Cheng B, Chen X. Estrogen accelerates G1 to S phase transition and induces a G2/M phase-predominant apoptosis in synthetic vascular smooth muscle cells. Int J Cardiol. 2007;118:381–8.  https://doi.org/10.1016/j.ijcard.2006.07.049.CrossRefPubMedGoogle Scholar
  33. 33.
    Wagner EF, Nebreda AR. Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 2009;9:537–49.  https://doi.org/10.1038/nrc2694.CrossRefPubMedGoogle Scholar
  34. 34.
    Lee CF, Qiao M, Schroder K, Zhao Q, Asmis R. Nox4 is a novel inducible source of reactive oxygen species in monocytes and macrophages and mediates oxidized low density lipoprotein-induced macrophage death. Circ Res. 2010;106:1489–97.  https://doi.org/10.1161/circresaha.109.215392.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Demiroglu-Zergeroglu A, Candemir G, Turhanlar E, Sagir F, Ayvali N. EGFR-dependent signalling reduced and p38 dependent apoptosis required by Gallic acid in malignant mesothelioma cells. Biomed Pharmacother Biomed Pharmacother. 2016;84:2000–7.  https://doi.org/10.1016/j.biopha.2016.11.005.CrossRefPubMedGoogle Scholar
  36. 36.
    Li H, Tian Z, Qu Y, Yang Q, Guan H, Shi B, Ji M, Hou P. SIRT7 promotes thyroid tumorigenesis through phosphorylation and activation of Akt and p70S6K1 via DBC1/SIRT1 axis. Oncogene. 2019;38:345–59.  https://doi.org/10.1038/s41388-018-0434-6.CrossRefPubMedGoogle Scholar
  37. 37.
    Reinhardt HC, Aslanian AS, Lees JA, Yaffe MB. p53-deficient cells rely on ATM- and ATR-mediated checkpoint signaling through the p38MAPK/MK2 pathway for survival after DNA damage. Cancer Cell. 2007;11:175–89.  https://doi.org/10.1016/j.ccr.2006.11.024.CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Freund A, Patil CK, Campisi J. p38MAPK is a novel DNA damage response-independent regulator of the senescence-associated secretory phenotype. EMBO J. 2011;30:1536–48.  https://doi.org/10.1038/emboj.2011.69.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Garcia-Cano J, Roche O, Cimas FJ, Pascual-Serra R, Ortega-Muelas M, Fernandez-Aroca DM, Sanchez-Prieto R. p38MAPK and chemotherapy: we always need to hear both sides of the story. Front Cell Dev Biol. 2016;4:69.  https://doi.org/10.3389/fcell.2016.00069.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Xue Q, Wang X, Wang P, Zhang K, Liu Q. Role of p38MAPK in apoptosis and autophagy responses to photodynamic therapy with Chlorin e6. Photodiagn Photodyn Ther. 2015;12:84–91.  https://doi.org/10.1016/j.pdpdt.2014.12.001.CrossRefGoogle Scholar
  41. 41.
    Ma, M.M.; Zhu, X.L.; Wang, L.; Hu, X.F.; Wang, Z.; Zhao, J.; Ma, Y.T.; Yang, Y.N.; Chen, B.D.; Liu, F. beta3-adrenoceptor impacts apoptosis in cultured cardiomyocytes via activation of PI3 K/Akt and p38MAPK. Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban 2016, 36, 1–7,  https://doi.org/10.1007/s11596-016-1533-7.CrossRefGoogle Scholar
  42. 42.
    Sun WJ, Huang H, He B, Hu DH, Li PH, Yu YJ, Zhou XH, Lv Z, Zhou L, Hu TY, et al. Romidepsin induces G2/M phase arrest via Erk/cdc25C/cdc2/cyclinB pathway and apoptosis induction through JNK/c-Jun/caspase3 pathway in hepatocellular carcinoma cells. Biochem Pharmacol. 2017;127:90–100.  https://doi.org/10.1016/j.bcp.2016.12.008.CrossRefPubMedGoogle Scholar
  43. 43.
    Kumar A, Singh UK, Kini SG, Garg V, Agrawal S, Tomar PK, Pathak P, Chaudhary A, Gupta P, Malik A. JNK pathway signaling: a novel and smarter therapeutic targets for various biological diseases. Future Med Chem. 2015;7:2065–86.  https://doi.org/10.4155/fmc.15.132.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Graduate SchoolTianjin Medical UniversityTianjinPeople’s Republic of China
  2. 2.Graduate SchoolTianjin University of Traditional Chinese MedicineTianjinPeople’s Republic of China
  3. 3.Departments of Colorectal SurgeryTianjin Union Medical CenterTianjinPeople’s Republic of China
  4. 4.Department of PathologyTianjin Union Medical CenterTianjinPeople’s Republic of China
  5. 5.Department of PathologyTianjin Nankai HospitalTianjinPeople’s Republic of China

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