Molecular and Cellular Biochemistry

, Volume 381, Issue 1–2, pp 61–68

p53siRNA therapy reduces cell proliferation, migration and induces apoptosis in triple negative breast cancer cells

  • Cornelia Braicu
  • Valentina Pileczki
  • Alexandru Irimie
  • Ioana Berindan-Neagoe
Article

Abstract

p53 protein is probably the best known tumor suppressor. Earlier reports proved that human breast cancer cells expressing mutant p53 displayed resistance to apoptosis. This study is intended to investigate, the potential applications of RNA interference (RNAi) to block p53 expression, as well as its subsequent effect on cell growth, apoptosis and migration on a triple negative human breast cancer cell line (Hs578T). p53siRNA significantly reduced cell index (CI) compared to the control and we observed an inhibition of cellular migration in the interval of time between 0 and 30 h, as shown in the data obtained by dynamic evaluation using the xCELLigence System. Also, by using PCR-array technology, a panel of 84 key genes involved in apoptosis was investigated. Our studies indicate that the knockdown of p53 expression by siRNA modulates several genes involved in cell death pathways and apoptosis, showing statistically significant gene expression differences for 22 genes, from which 18 were upregulated and 4 were downregulated. The present research also emphasizes the important role of BCL-2 pro-apoptotic family of genes (Bim, Bak, and Bax) in activating apoptosis and reducing cell proliferation by p53siRNA treatment. Death receptors cooperate with BCL-2 pro-apoptotic genes in reducing cell proliferation. The limited success may be due to the activation of the antiapoptotic gene Mcl-1, and it may be associated with the resistance of triple negative breast cancer cells to cancer treatment. Thus, targeting p53siRNA pathways using siRNA may serve as a promising therapeutic strategy for the treatment of breast cancers.

Keywords

p53siRNA Triple negative breast cells Cell proliferation Migration Apoptosis 

Supplementary material

11010_2013_1688_MOESM1_ESM.pptx (252 kb)
Supplementary material 1 (PPTX 252 kb)

References

  1. 1.
    Wang Z, Sun Y (2010) Targeting p53 for novel anticancer therapy. Transl Oncol 3(1):1–12PubMedGoogle Scholar
  2. 2.
    Finlay CA, Hinds PW, Levine AJ (1989) The p53 proto-oncogene can act as a suppressor of transformation. Cell 57(7):1083–1093PubMedCrossRefGoogle Scholar
  3. 3.
    Bishayee A, Ahmed S, Brankov N, Perloff M (2011) Triterpenoids as potential agents for the chemoprevention and therapy of breast cancer. Front Biosci 16:980–996PubMedCrossRefGoogle Scholar
  4. 4.
    Rahman M, Davis SR, Pumphrey JG, Bao J, Nau MM, Meltzer PS, Lipkowitz S (2009) TRAIL induces apoptosis in triple-negative breast cancer cells with a mesenchymal phenotype. Breast Cancer Res Treat 113(2):217–230PubMedCrossRefGoogle Scholar
  5. 5.
    Liu Y-Y, Patwardhan G, Bhinge K, Gupta V, Gu X, Jazwinski M (2011) Suppression of glucosylceramide synthase restores p53-dependent apoptosis in mutant p53 cancer cells. Cancer Res 71(6):2276–2285PubMedCrossRefGoogle Scholar
  6. 6.
    Norberg T, Klaar S, Karf G, Nordgren H, Holmberg L, Bergh J (2001) Increased p53 mutation frequency during tumor progression––results from a breast cancer cohort. Cancer Res 61:8317–8321PubMedGoogle Scholar
  7. 7.
    Sigal A, Rotter V (2000) Oncogenic mutations of the p53 tumor suppressor: the demons of the guardian of the genome. Cancer Res 60:6788–6793PubMedGoogle Scholar
  8. 8.
    Vikhanskaya F, Lee K, Mazzoletti M, Broggini M, Sabapathy K (2007) Cancer-derived p53 mutants suppress p53-target gene expression—potential mechanism for gain of function of mutant p53. Nucleic Acids Res 35(6):2093–2104PubMedCrossRefGoogle Scholar
  9. 9.
    Kim DH, Rossi JJ (2007) Strategies for silencing human disease using RNA interference. Nat Rev Genet 8:173–184PubMedCrossRefGoogle Scholar
  10. 10.
    Brummelkamp TR, Bernards R, Agami R (2002) Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell 2:243–247PubMedCrossRefGoogle Scholar
  11. 11.
    Kim DH, Rossi JJ (2007) Strategies for silencing human disease using RNA interference. Nat Rev Genet 8:173–184PubMedCrossRefGoogle Scholar
  12. 12.
    Rye PD, Stigbrand T (2004) Interfering with cancer: A brief outline of advances in RNA interference in oncology. Tumor Biology 25:5–6Google Scholar
  13. 13.
    Lacroix M, Toillon RA, Leclercq G (2006) p53 and breast cancer, an update. Endocr Relat Cancer 13(2):293–325PubMedCrossRefGoogle Scholar
  14. 14.
    Gasco M, Shami S, Crook T (2002) The p53 pathway in breast cancer. Breast Cancer Res 4:70–76PubMedCrossRefGoogle Scholar
  15. 15.
    Ma CX, Cai S, Li S, Ryan CE, Guo Z, Schaiff WT, Lin L, Hoog J, Goiffon RJ, Prat A, Aft RL, Ellis MJ, Piwnica-Worms H (2012) Targeting Chk1 in p53-deficient triple-negative breast cancer is therapeutically beneficial in human-in-mouse tumor Models. J Clin Invest 122(4):1541–1552PubMedCrossRefGoogle Scholar
  16. 16.
    Zhao R, Gish K, Murphy M, Yin Y, Notterman D, Hoffman WH, Tom E, Mack DH, Levine AJ (2000) Analysis of p53-regulated gene expression patterns using oligonucleotide arrays. Genes Dev 14:981–993PubMedCrossRefGoogle Scholar
  17. 17.
    Zhou M, Liu Z, Zhao Y, Ding Y, Liu H, Xi Y, Xiong W, Li G, Lu J, Fodstad O, Riker AI, Tan M (2010) MicroRNA-125b confers the resistance of breast cancer cells to paclitaxel through suppression of pro-apoptotic Bcl-2 antagonist killer 1 (Bak1) expression. J Biol Chem 285(28):21496–21507PubMedCrossRefGoogle Scholar
  18. 18.
    Scott GB, Bowles PA, Wilson EB, Meade JL, Low BC, Davison A, Davison A, Blair GE, Cook GP (2010) Identification of the BCL2/adenovirus E1B–19K protein-interacting protein 2 (BNIP-2) as a granzyme B target during human natural killer cell-mediated killing. Biochem J 431:423–431PubMedGoogle Scholar
  19. 19.
    Morel C, Carlson SM, White FM, Davis RJ (2009) Mcl-1 integrates the opposing actions of signaling pathways that mediate survival and apoptosis. Mol Cell Biol 29(14):3845–3852PubMedCrossRefGoogle Scholar
  20. 20.
    Plati J, Bucur O, Khosravi-Far R (2011) Apoptotic cell signaling in cancer progression and therapy. Integr Biol (Camb) 3(4):279–296CrossRefGoogle Scholar
  21. 21.
    Bruey JM, Bruey-Sedano N, Luciano F, Zhai D, Balpai R, Xu C, Kress CL, Bailly-Maitre B, Li X, Osterman A, Mastsuzawa S, Terskikh AV, Faustin B, Reed JC (2007) Bcl-2 and Bcl-XL regulate proinflammatory caspase-1 activation by interaction with NALP1. Cell 129:45–56PubMedCrossRefGoogle Scholar
  22. 22.
    Aggarwal BB, Shishodia S, Sandur SK, Pandey MK, Sethi G (2006) Inflammation and cancer: how hot is the link? Biochem Pharmacol 72(11):1605–1621PubMedCrossRefGoogle Scholar
  23. 23.
    Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35(4):495–516PubMedCrossRefGoogle Scholar
  24. 24.
    Booy EP, Henson ES, Gibson SB (2011) Epidermal growth factor regulates Mcl-1 expression through the MAPK-Elk-1 signalling pathway contributing to cell survival in breast cancer. Oncog 30:2367–2378Google Scholar
  25. 25.
    Woods NT, Yamaguchi H, Lee FY, Bhalla KN (2007) Wang Anoikis HG. initiated by Mcl-1 degradation and Bim induction, is deregulated during oncogenesis. Cancer Res 67:10744Google Scholar
  26. 26.
    Qin W, Hu J, Guo M, Xu J, Li J, Yao G et al (2003) BNIPL-2, a novel homologue of BNIP-2, interacts with Bcl-2 and Cdc42GAP in apoptosis. Biochem Biophys Res Commun 308:379–385Google Scholar
  27. 27.
    Debily MA, Marhomy SE, Boulanger V, Eveno E, Mariage-Samson R, Camarca A et al (2009) A functional and regulatory network associated with PIP expression in human breast cancer. PLoS One 4(3):4696Google Scholar
  28. 28.
    Lu Y, Chen GQ (2011) Effector caspases and leukemia. Int J of Cell Biol. doi:10.1155/2011/738301
  29. 29.
    Valentín-Acevedo A, Sinquett FL, Covey LR (2011) c-Rel deficiency increases caspase-4 expression and leads to ER stress and necrosis in EBV-transformed cells. PLoS One 6(10):25467Google Scholar
  30. 30.
    Smolnikar K, Löffek S, Schulz T, Michna H, Diel P (2000) Treatment with the pure antiestrogen faslodex (ICI 182780) induces tumor necrosis factor receptor 1 (TNFR1) expression in MCF-7 breast cancer cells. Breast Cancer Res Treat 63(3):249–259Google Scholar
  31. 31.
    Adams JM, Cory S (2007) The Bcl-2 apoptotic switch in cancer development and therapy Bcl-2 apoptotic switch in cancer. Oncog 26:1324–1337Google Scholar
  32. 32.
    Miller LD, Smeds J, George J, Vega VB, Vergara L, Ploner A (2005) An expression signature for p53 status in human breast cancer predicts mutation status, transcriptional effects, and patient survival. PNAS 2(38):13550–13555Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Cornelia Braicu
    • 1
    • 2
  • Valentina Pileczki
    • 1
  • Alexandru Irimie
    • 3
    • 4
  • Ioana Berindan-Neagoe
    • 1
    • 2
    • 5
  1. 1.Research Center for Functional Genomics, Biomedicine and Translational Medicine“Iuliu Hatieganu” University of Medicine and PharmacyCluj-NapocaRomania
  2. 2.Department of Functional Genomics and Experimental PathologyThe Oncological Institute “Prof. Dr. Ion Chiricuta”Cluj-NapocaRomania
  3. 3.Department of Surgical Oncology“I. Hatieganu” University of Medicine and PharmacyCluj-NapocaRomania
  4. 4.Department of SurgeryThe Oncological Institute“Prof. Dr.Ion Chiricuta”Cluj-NapocaRomania
  5. 5.Department of Immunology“Iuliu Hatieganu” University of Medicine and PharmacyCluj-NapocaRomania

Personalised recommendations