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Tumor Biology

, Volume 37, Issue 10, pp 13225–13235 | Cite as

Depletion of UBA protein 2-like protein inhibits growth and induces apoptosis of human colorectal carcinoma cells

  • Rui Chai
  • Xiaojun Yu
  • Shiliang Tu
  • Bo’an Zheng
Original Article

Abstract

Ubiquitin-proteasome system regulates cell proliferation, apoptosis, angiogenesis, and motility, which are processes with particular importance for carcinogenesis. UBA protein 2-like protein (UBAP2L) was found to be associated with proteasome; however, its biological function is largely unknown. In this study, the mRNA levels of UBAP2L in human normal and colorectal carcinoma tissues were analyzed using the datasets from the publicly available Oncomine database (www.oncomine.org) and found UBAP2L was overexpressed in colorectal carcinoma tissues. Furthermore, we elucidated the role of UBAP2L in human colorectal cancer via an RNA interference lentivirus system in three colorectal carcinoma cell lines HCT116, SW1116, and RKO. Knockdown of UBAP2L led to suppressed cell proliferation and impaired colony formation. UBAP2L depletion in HCT116 and RKO cells also induced cell cycle arrest as well as apoptosis. Moreover, the phosphorylation of PRAS40, Bad, and the cleavage of PARP were remarkably increased after UBAP2L knockdown by Intracellular signaling array and also the activation of P38 was obviously decreased and the cleavage of Caspase 3 and Bax were increased after UBAP2L silencing by western blot assay, indicated that UBAP2L might be involved in the cell growth by the regulation of apoptosis-related proteins. Our findings indicated that UBAP2L may be essential for colorectal carcinoma growth and survival. Lentivirus-mediated small interfering RNA against UBAP2L might serve as a potential therapeutic approach for the treatment of colorectal cancer.

Keywords

Apoptosis Colorectal cancer Proliferation siRNA UBAP2L 

Notes

Acknowledgments

The authors are thankful for the financial support from Natural Science Foundation of Zhejiang province (grant no. Y15H160152).

Compliance with ethical standards

Conflicts of interest

None

References

  1. 1.
    Hershko A. The ubiquitin system for protein degradation and some of its roles in the control of the cell division cycle. Cell Death Differ. 2005;12:1191–7.CrossRefPubMedGoogle Scholar
  2. 2.
    Koegl M, Hoppe T, Schlenker S, Ulrich HD, et al. A novel ubiquitination factor, e4, is involved in multiubiquitin chain assembly. Cell. 1999;96:635–44.CrossRefPubMedGoogle Scholar
  3. 3.
    Voutsadakis IA. The ubiquitin-proteasome system in colorectal cancer. Biochim Biophys Acta. 2008;1782:800–8.CrossRefPubMedGoogle Scholar
  4. 4.
    Micel LN, Tentler JJ, Smith PG, Eckhardt GS. Role of ubiquitin ligases and the proteasome in oncogenesis: novel targets for anticancer therapies. J Clin Oncol. 2013;31:1231–8.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Mani A, Gelmann EP. The ubiquitin-proteasome pathway and its role in cancer. J Clin Oncol. 2005;23:4776–89.CrossRefPubMedGoogle Scholar
  6. 6.
    Duan DR, Pause A, Burgess WH, Aso T, et al. Inhibition of transcription elongation by the vhl tumor suppressor protein. Science. 1995;269:1402–6.CrossRefPubMedGoogle Scholar
  7. 7.
    Werness BA, Levine AJ, Howley PM. Association of human papillomavirus types 16 and 18 e6 proteins with p53. Science. 1990;248:76–9.CrossRefPubMedGoogle Scholar
  8. 8.
    Clevers H. Wnt breakers in colon cancer. Cancer Cell. 2004;5:5–6.CrossRefPubMedGoogle Scholar
  9. 9.
    Ilyas M. Wnt signalling and the mechanistic basis of tumour development. J Pathol. 2005;205:130–44.CrossRefPubMedGoogle Scholar
  10. 10.
    Fodde R, Smits R, Clevers H. Apc, signal transduction and genetic instability in colorectal cancer. Nat Rev Cancer. 2001;1:55–67.CrossRefPubMedGoogle Scholar
  11. 11.
    Coutts AS, La Thangue NB. The p53 response: emerging levels of co-factor complexity. Biochem Biophys Res Commun. 2005;331:778–85.CrossRefPubMedGoogle Scholar
  12. 12.
    Xu J, Attisano L. Mutations in the tumor suppressors smad2 and smad4 inactivate transforming growth factor beta signaling by targeting smads to the ubiquitin-proteasome pathway. Proc Natl Acad Sci U S A. 2000;97:4820–5.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Fang JY, Richardson BC. The mapk signalling pathways and colorectal cancer. Lancet Oncol. 2005;6:322–7.CrossRefPubMedGoogle Scholar
  14. 14.
    Vivanco I, Sawyers CL. The phosphatidylinositol 3-kinase akt pathway in human cancer. Nat Rev Cancer. 2002;2:489–501.CrossRefPubMedGoogle Scholar
  15. 15.
    Buchberger A. From uba to ubx: new words in the ubiquitin vocabulary. Trends Cell Biol. 2002;12:216–21.CrossRefPubMedGoogle Scholar
  16. 16.
    Madura K. The ubiquitin-associated (uba) domain: on the path from prudence to prurience. Cell Cycle. 2002;1:235–44.CrossRefPubMedGoogle Scholar
  17. 17.
    Andersen KM, Hofmann K, Hartmann-Petersen R. Ubiquitin-binding proteins: similar, but different. Essays Biochem. 2005;41:49–67.CrossRefPubMedGoogle Scholar
  18. 18.
    Hurley JH, Lee S, Prag G. Ubiquitin-binding domains. Biochem J. 2006;399:361–72.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Hicke L, Schubert HL, Hill CP. Ubiquitin-binding domains. Nat Rev Mol Cell Biol. 2005;6:610–21.CrossRefPubMedGoogle Scholar
  20. 20.
    Hofmann K, Bucher P. The uba domain: a sequence motif present in multiple enzyme classes of the ubiquitination pathway. Trends Biochem Sci. 1996;21:172–3.CrossRefPubMedGoogle Scholar
  21. 21.
    Andersson HA, Passeri MF, Barry MA. Rad23 as a reciprocal agent for stimulating or repressing immune responses. Hum Gene Ther. 2005;16:634–41.CrossRefPubMedGoogle Scholar
  22. 22.
    Lee JJ, Kim YM, Jeong J, Bae DS, et al. Ubiquitin-associated (uba) domain in human fas associated factor 1 inhibits tumor formation by promoting hsp70 degradation. PLoS One. 2012;7:e40361.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Wilde IB, Brack M, Winget JM, Mayor T. Proteomic characterization of aggregating proteins after the inhibition of the ubiquitin proteasome system. J Proteome Res. 2011;10:1062–72.CrossRefPubMedGoogle Scholar
  24. 24.
    Sawyer JR. The prognostic significance of cytogenetics and molecular profiling in multiple myeloma. Cancer Genet. 2011;204:3–12.CrossRefPubMedGoogle Scholar
  25. 25.
    Zhu ZZ, Wang D, Cong WM, Jiang H, et al. Sex-related differences in DNA copy number alterations in hepatitis b virus-associated hepatocellular carcinoma. Asian Pac J Cancer Prev. 2012;13:225–9.CrossRefPubMedGoogle Scholar
  26. 26.
    Naz RK, Dhandapani L. Identification of human sperm proteins that interact with human zona pellucida3 (zp3) using yeast two-hybrid system. J Reprod Immunol. 2010;84:24–31.CrossRefPubMedGoogle Scholar
  27. 27.
    Li D, Huang Y. Knockdown of ubiquitin associated protein 2-like inhibits the growth and migration of prostate cancer cells. Oncol Rep. 2014;32:1578–84.PubMedGoogle Scholar
  28. 28.
    Zhao B, Zong G, Xie Y, Li J, et al. Downregulation of ubiquitin-associated protein 2-like with a short hairpin RNA inhibits human glioma cell growth in vitro. Int J Mol Med. 2015;36:1012–8.PubMedPubMedCentralGoogle Scholar
  29. 29.
    McManus MT, Sharp PA. Gene silencing in mammals by small interfering RNAs. Nat Rev Genet. 2002;3:737–47.CrossRefPubMedGoogle Scholar
  30. 30.
    Devi GR. siRNA-based approaches in cancer therapy. Cancer Gene Ther. 2006;13:819–29.CrossRefPubMedGoogle Scholar
  31. 31.
    Skrzypczak M, Goryca K, Rubel T, Paziewska A, et al. Modeling oncogenic signaling in colon tumors by multidirectional analyses of microarray data directed for maximization of analytical reliability. PLoS One. 2010;5.Google Scholar
  32. 32.
    Hong Y, Downey T, KW E, Koh PK, et al. A ‘metastasis-prone’ signature for early-stage mismatch-repair proficient sporadic colorectal cancer patients and its implications for possible therapeutics. Clin Exp Metastasis. 2010;27:83–90.CrossRefPubMedGoogle Scholar
  33. 33.
    Gaedcke J, Grade M, Jung K, Camps J, et al. Mutated kras results in overexpression of dusp4, a map-kinase phosphatase, and smyd3, a histone methyltransferase, in rectal carcinomas. Genes Chromosom Cancer. 2010;49:1024–34.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Thedieck K, Polak P, Kim ML, Molle KD, et al. Pras40 and prr5-like protein are new mtor interactors that regulate apoptosis. PLoS One. 2007;2:e1217.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Konishi Y, Lehtinen M, Donovan N, Bonni A. Cdc2 phosphorylation of bad links the cell cycle to the cell death machinery. Mol Cell. 2002;9:1005–16.CrossRefPubMedGoogle Scholar
  36. 36.
    Bhatia M, Kirkland JB, Meckling-Gill KA. Overexpression of poly(adp-ribose) polymerase promotes cell cycle arrest and inhibits neutrophilic differentiation of nb4 acute promyelocytic leukemia cells. Cell Growth Differ. 1996;7:91–100.PubMedGoogle Scholar
  37. 37.
    Wang M, Medeiros BC, Erba HP, DeAngelo DJ, et al. Targeting protein neddylation: a novel therapeutic strategy for the treatment of cancer. Expert Opin Ther Targets. 2011;15:253–64.CrossRefPubMedGoogle Scholar
  38. 38.
    Soucy TA, Smith PG, Milhollen MA, Berger AJ, et al. An inhibitor of nedd8-activating enzyme as a new approach to treat cancer. Nature. 2009;458:732–6.CrossRefPubMedGoogle Scholar
  39. 39.
    Yi-Zhuo L, An-Mei D, Liang-Hui L, Guo-Yan L, et al. Prognostic role of phospho-pras40 (thr246) expression in gastric cancer. Arch Med Sci. 2014;10:149–53.Google Scholar
  40. 40.
    Marchion DC, Cottrill HM, Xiong Y, Chen N, et al. Bad phosphorylation determines ovarian cancer chemosensitivity and patient survival. Clin Cancer Res. 2011;17:6356–66.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Chon HS, Marchion DC, Xiong Y, Chen N, et al. The bcl2 antagonist of cell death pathway influences endometrial cancer cell sensitivity to cisplatin. Gynecol Oncol. 2012; 124:119–24.Google Scholar
  42. 42.
    Bressenot A, Marchal S, Bezdetnaya L, Garrier J, et al. Assessment of apoptosis by immunohistochemistry to active caspase-3, active caspase-7, or cleaved parp in monolayer cells and spheroid and subcutaneous xenografts of human carcinoma. J Histochem Cytochem. 2009;57:289–300.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Cano E, Mahadevan LC. Parallel signal processing among mammalian mapks. Trends Biochem Sci. 1995;20:117–22.CrossRefPubMedGoogle Scholar
  44. 44.
    Ono K, Han J. The p38 signal transduction pathway: activation and function. Cell Signal. 2000;12:1–13.CrossRefPubMedGoogle Scholar
  45. 45.
    Chen L, Mayer JA, Krisko TI, Speers CW, et al. Inhibition of the p38 kinase suppresses the proliferation of human er-negative breast cancer cells. Cancer Res. 2009;69:8853–61.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Kim JY, Park JH. Ros-dependent caspase-9 activation in hypoxic cell death. FEBS Lett. 2003;549:94–8.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Rui Chai
    • 1
  • Xiaojun Yu
    • 2
  • Shiliang Tu
    • 1
  • Bo’an Zheng
    • 1
  1. 1.Department of Colorectal SurgeryZhejiang Provincial People’s HospitalHangzhouChina
  2. 2.Department of Gastroenterological SurgeryZhejiang Provincial People’s HospitalHangzhouChina

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