Article | Published:

TC2N, a novel oncogene, accelerates tumor progression by suppressing p53 signaling pathway in lung cancer

Abstract

The protein containing the C2 domain has been well documented for its essential roles in endocytosis, cellular metabolism and cancer. Tac2-N (TC2N) is a tandem C2 domain-containing protein, but its function, including its role in tumorigenesis, remains unknown. Here, we first identified TC2N as a novel oncogene in lung cancer. TC2N was preferentially upregulated in lung cancer tissues compared with adjacent normal lung tissues. High TC2N expression was significantly associated with poor outcome of lung cancer patients. Knockdown of TC2N markedly induces cell apoptosis and cell cycle arrest with repressing proliferation in vitro, and suppresses tumorigenicity in vivo, whereas overexpression of TC2N has the opposite effects both in vitro and in vivo. Using a combination of TCGA database and bioinformatics, we demonstrate that TC2N is involved in regulation of the p53 signaling pathway. Mechanistically, TC2N attenuates p53 signaling pathway through inhibiting Cdk5-induced phosphorylation of p53 via inducing Cdk5 degradation or disrupting the interaction between Cdk5 and p53. Moreover, the blockade of p53 attenuates the function of TC2N knockdown in the regulation of cell proliferation and apoptosis. In addition, downregulated TC2N is involved in the apoptosis of lung cancer cells induced by doxorubicin, leading to p53 pathway activation. Overall, these findings uncover a role for the p53 inactivator TC2N in regulating the proliferation and apoptosis of lung cancer cells. Our present study provides novel insights into the mechanism of tumorigenesis in lung cancer.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Torre LA1, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.

  2. 2.

    Yuan S, Yu Z, Liu Q, Zhang M, Xiang Y, Wu N, et al. GPC5, a novel epigenetically silenced tumor suppressor, inhibits tumor growth by suppressing Wnt/beta-catenin signaling in lung adenocarcinoma. Oncogene. 2016;35:6120–31.

  3. 3.

    Neri S, Yoshida J, Ishii G, Matsumura Y, Aokage K, Hishida T, et al. Prognostic impact of microscopic vessel invasion and visceral pleural invasion in non-small cell lung cancer: a retrospective analysis of 2657 patients. Ann Surg. 2014;260:383–8.

  4. 4.

    Zhang J, Gold KA, Lin HY, Swisher SG, Xing Y, Lee JJ, et al. Relationship between tumor size and survival in non-small-cell lung cancer (NSCLC): an analysis of the surveillance, epidemiology, and end results (SEER) registry. J Thorac Oncol. 2015;10:682–90.

  5. 5.

    Wanders R, Steevens J, Botterweck A, Dingemans AM, Reymen B, Av Baardwijk, et al. Treatment with curative intent of stage III non-small cell lung cancer patients of 75 years: a prospective population-based study. Eur J Cancer. 2011;47:2691–2197.

  6. 6.

    Fukuda M, Mikoshiba K. Tac2-N, an atypical C-type tandem C2 protein localized in the nucleus. FEBS Lett. 2001;503:217–8.

  7. 7.

    Duncan RR, Shipston MJ, Chow RH. Double C2 protein. A review. Biochimie. 2000;82:421–6.

  8. 8.

    Corbalan-Garcia S, Gómez-Fernández JC. Signaling through C2 domains: more than one lipid target. Biochim Biophys Acta. 2014;1838:1536–47.

  9. 9.

    Cho W, Stahelin RV. Membrane binding and subcellular targeting of C2 domains. Biochim Biophys Acta. 2006;1761:838–49.

  10. 10.

    Farah CA, Sossin WS. The role of C2 domains in PKC signaling. Adv Exp Med Biol. 2012;740:663–83.

  11. 11.

    Kabekkodu SP, Bhat S, Radhakrishnan R, Aithal A, Mascarenhas R, Pandey D, et al. DNA promoter methylation-dependent transcription of the double C2-like domain β (DOC2B) gene regulates tumor growth in human cervical cancer. J Biol Chem. 2014;289:10637–49.

  12. 12.

    Blomme A, Costanza B, de Tullio P, Thiry M, Van Simaeys G, Boutry S, et al. Myoferlin regulates cellular lipid metabolism and promotes metastases in triple-negative breast cancer. Oncogene. 2017;36:2116–30.

  13. 13.

    Yue X, Zhao Y, Xu Y, Zheng M, Feng Z, Hu W. Mutant p53 in cancer: Accumulation, gain-of-function and therapy. J Mol Biol. 2017;429:1595–606.

  14. 14.

    Liu G, Chen X. Regulation of the p53 transcriptional activity. J Cell Biochem. 2006;97:448–58.

  15. 15.

    Zhang J, Krishnamurthy PK, Johnson GV. Cdk5 phosphorylates p53 and regulates its activity. J Neurochem. 2002;81:307–13.

  16. 16.

    Lee JH, Kim HS, Lee SJ, Kim KT. Stabilization and activation of p53 induced by Cdk5 contributes to neuronal cell death. J Cell Sci. 2007;120:2259–71.

  17. 17.

    Ding F, Xiao H, Wang M, Xie X, Hu F. The role of the ubiquitin-proteasome pathway in cancer development and treatment. Front Biosci. 2014;19:886–95.

  18. 18.

    Xu Y1, Diao Y, Qi S, Pan X, Wang Q, Xin Y, et al. Phosphorylated Hsp27 activates ATM-dependent p53 signaling and mediates the resistance of MCF-7 cells to doxorubicin-induced apoptosis. Cell Signal. 2013;25:1176–85.

  19. 19.

    Bernatchez PN, Acevedo L, Fernandez-Hernando C, Murata T, Chalouni C, Kim J, et al. Myoferlin regulates vascular endothelial growth factor receptor-2 stability and function. J Biol Chem. 2007;282:30745–53.

  20. 20.

    Demonbreun AR, Posey AD, Heretis K, Swaggart KA, Earley JU, Pytel P, et al. Myoferlin is required for insulin-like growth factor response and muscle growth. FASEB J. 2010;24:1284–95.

  21. 21.

    Turtoi A1, Blomme A, Bellahcène A, Gilles C, Hennequière V, Peixoto P, et al. Myoferlin is a key regulator of EGFR activity in breast cancer. Cancer Res. 2013;73:5438–48.

  22. 22.

    Yu C, Sharma A, Trane A, Utokaparch S, Leung C, Bernatchez P. Myoferlin gene silencing decreases Tie-2 expression in vitro and angiogenesis in vivo. Vasc Pharmacol. 2011;55:26–33.

  23. 23.

    Bernatchez PN, Sharma A, Kodaman P, Sessa WC. Myoferlin is critical for endocytosis in endothelial cells. Am J Physiol Cell Physiol. 2009;297:484–92.

  24. 24.

    Fahmy K, Gonzalez A, Arafa M, Peixoto P, Bellahcène A, Turtoi A, et al. Myoferlin plays a key role in VEGFA secretion and impacts tumor-associated angiogenesis in human pancreas cancer. Int J Cancer. 2016;138:652–63.

  25. 25.

    Iyoda A, Makino T, Koezuka S, Otsuka H, Hata Y. Treatment options for patients with large cell neuroendocrine carcinoma of the lung. Gen Thorac Cardiovasc Surg. 2014;62:351–6.

  26. 26.

    Sakurai H1, Asamura H. Large-cell neuroendocrine carcinoma of the lung: surgical management. Thorac Surg Clin. 2014;24:305–11.

  27. 27.

    Scudla V, Ordeltova M, Bacovsky J, Vytrasova M, Horak P, Minarik J. The relationship between proliferation and apoptosis in patients with monoclonal gammopathy of undetermined significance or multiple myeloma. Haematologica. 2005;90:1713–4.

  28. 28.

    Efeyan A, Serrano M. p53: guardian of the genome and policeman of the oncogenes. Cell Cycle. 2007;6:1006–10.

  29. 29.

    Haupt Y, Maya R, Kazaz A, Oren M. Mdm2 promotes the rapid degradation of p53. Nature. 1997;387:296–9.

  30. 30.

    Kubbutat MH, Jones SN, Vousden KH. Regulation of p53 stability by Mdm2. Nature. 1997;387:299–303.

  31. 31.

    Vogelstein B, Lane D, Levine AJ. Surfing the p53 network. Nature. 2000;408:307–10.

  32. 32.

    Miyashita T, Reed JC. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell. 1995;80:293–9.

  33. 33.

    el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, et al. WAF1, a potential mediator of p53 tumor suppression. Cell. 1993;75:817–25.

  34. 34.

    Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell. 1993;75:805–16.

  35. 35.

    Haldar S, Negrini M, Monne M, Sabbioni S, Croce CM. Down-regulation of bcl-2 by p53 in breast cancer cells. Cancer Res. 1994;54:2095–7.

  36. 36.

    Oda E, Ohki R, Murasawa H, Nemoto J, Shibue T, Yamashita T, et al. Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis. Science. 2000;288:1053–8.

  37. 37.

    Horn HF, Vousden KH. Coping with stress: multiple ways to activate p53. Oncogene. 2007;26:1306–16.

  38. 38.

    Takenaka I, Morin F, Seizinger BR, Kley N. Regulation of the sequence-specific DNA binding function of p53 by protein kinase C and protein phosphatases. J Biol Chem. 1995;270:5405–11.

  39. 39.

    Wang Y, Prives C. Increased and altered DNA binding of human p53 by S and G2/M but not Gl cyclin-dependent kinases. Nature. 1995;376:88–91.

  40. 40.

    Lu H, Fisher RP, Bailey P, Levine AJ. The CDK7-cycH-p36 complex of transcription factor IIH phosphorylates p53, enhancing its sequence-specific DNA binding activity in vitro. Mol Cell Biol. 1997;17:5923–34.

  41. 41.

    Waterman MJ, Stavridi ES, Waterman JL, Halazonetis TD. ATM-dependent activation of p53 involves dephosphorylation and association with 14-3-3 proteins. Nat Genet. 1998;19:175–8.

  42. 42.

    Ajay AK, Upadhyay AK, Singh S, Vijayakumar MV, Kumari R, Pandey V, et al. Cdk5 phosphorylates non-genotoxically overexpressed p53 following inhibition of PP2A to induce cell cycle arrest/apoptosis and inhibits tumor progression. Mol Cancer. 2010;9:204.

  43. 43.

    Gibbons DL, Byers LA, Kurie JM. Smoking, p53 mutation, and lung cancer. Mol Cancer Res. 2014;12:3–13.

  44. 44.

    Molina-Vila MA, Bertran-Alamillo J, Gascó A, Mayo-de-las-Casas C, Sánchez-Ronco M, Pujantell-Pastor L, et al. Nondisruptive p53 mutations are associated with shorter survival in patients with advanced non-small cell lung cancer. Clin Cancer Res. 2014;20:4647–59.

  45. 45.

    Tan BS, Tiong KH, Choo HL, Chung FF, Hii LW, Tan SH, et al. Mutant p53-R273H mediates cancer cell survival and anoikis resistance through AKT-dependent suppression of BCL2-modifying factor (BMF). Cell Death Dis. 2015;6:e1826.

  46. 46.

    Dong P, Tada M, Hamada J, Nakamura A, Moriuchi T, Sakuragi N. p53 dominant-negative mutant R273H promotes invasion and migration of human endometrial cancer HHUA cells. Clin Exp Metastas-. 2007;24:471–83.

  47. 47.

    Han F, Liu W, Jiang X, Shi X, Yin L, Ao L, et al. SOX30, a novel epigenetic silenced tumor suppressor, promotes tumor cell apoptosis by transcriptional activating p53 in lung cancer. Oncogene. 2015;34:4391–402.

  48. 48.

    Liu WB, Han F, Du XH, Jiang X, Li YH, Liu Y, et al. Epigenetic silencing of Aristaless-like homeobox-4, a potential tumor suppressor gene associated with lung cancer. Int J Cancer. 2014;134:1311–22.

  49. 49.

    Hu X, Wang L, Sun W, Xiao L, Wu Y, Zhuo Y, et al. AP-2β enhances p53-mediated transcription of the αB-crystallin gene through stabilizing p53. Mol Biol Rep. 2012;39:209–14.

  50. 50.

    Lv J, Zhu P, Yang Z, Li M, Zhang X, Cheng J, et al. PCDH20 functions as a tumour-suppressor gene through antagonizing the Wnt/β-catenin signalling pathway in hepatocellular carcinoma. J Viral Hepat. 2015;22:201–11.

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 81773461 and 81573179). We would like to thank all patients involved in this study. The authors would like to thank American Journal Experts for language editing.

Author contributions

X-LH, FH, JC, LA, W-BL and J-YL were responsible for the experimental design. X-LH, NZ, D-DW and J-PC contributed to the execution of experiments, data statistics and manuscript composition. FH, NZ, H-QC and J-YL participated in performing the experiment and in the manuscript mapping and submission. X-LH, FH, W-BL, LY, XJ, Z-HC and J-YL participated in the discussion and interpretation of data. FH, LY, XJ, JC and J-YL conceived the study and revised the manuscript. FH, W-BL, JC and J-YL were responsible for the funding application, and the supervision and management of the project. All authors have contributed to and approved the final manuscript.

Author information

Correspondence to Jia Cao or Jin-yi Liu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Edited by H.-U. Simon

Electronic supplementary material

supplement information

Supplementary Figure S1

Supplementary Figure S2

Supplementary Figure S3

Supplementary Figure S4

Supplementary Figure S5

Supplementary Figure S6

Dataset 1

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8