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Tumor Biology

, Volume 37, Issue 1, pp 1159–1171 | Cite as

Targeting polyamine biosynthetic pathway through RNAi causes the abrogation of MCF 7 breast cancer cell line

  • Enna Dogra Gupta
  • Manendra Pachauri
  • Prahlad Chandra Ghosh
  • Manchikatla Venkat Rajam
Original Article

Abstract

The diamine putrescine and polyamines, spermidine (triamine) and spermine (tetraamine) are small organic polycations that play an indispensable role in key cellular processes such as the regulation of growth, differentiation, and macromolecular functions. Elevated levels of polyamines (PAs) have been shown to be one of the major factors involved in carcinogenesis. In this study, specific silencing of the expression of three genes of PA biosynthesis pathway, ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC), and spermidine synthase (SPDSYN) was achieved using RNA interference in MCF 7 breast cancer cell line. For optimizing the effective small interfering nucleic acid (siNA), three variants of ODC siNA [siRNA, locked nucleic acid (LNA)-modified siRNA, and siHybrid (RNA and DNA hybrid)] were used and a dose- and time-dependent study was conducted. The PA biosynthetic genes were targeted individually and in combination. RNAi-mediated reduction in the expression of PA biosynthesis genes resulted in distorted cell morphology, reduced cancer cell viability, and migration characteristic. The most promising results were observed with the combined treatment of siSPDSYN and siODC with 83 % cell growth inhibition. On analyzing the messenger RNA (mRNA) expression profile of the cell cycle and apoptosis-related genes, it was observed that RNAi against PA biosynthetic genes downregulated the expression of CDK8, CCNE2, CCNH, CCNT1, CCNT2, CCNF, PCNA, CCND1, and CDK2, and upregulated the expression of E2F4, BAX, FAS, TP53, CDKN1A, BAK1, CDKN1B, ATM, GRANB, and ATR genes when compared with control-transfected cells. These results suggest that the targeting polyamine biosynthesis through RNAi approach could be a promising strategy for breast cancer therapy and might be extended for therapy of other cancers.

Keywords

Polyamines Cell division Cancer RNA interference Polyamine biosynthesis genes 

Notes

Acknowledgments

E.D.G. and M.P. are supported by research fellowships from Council of Scientific and Industrial Research (CSIR) and University Grants Commission (UGC), Government of India, respectively. This work was supported by Department of Science and Technology (DST - Grant (no. SR/SO/BB-29/2008) to M.V.R. and P.C.G. We acknowledge Prof. Alo Nag for providing MCF 7 cell line. M. V. R is thankful to DST for FIST (Level II) program, UGC for SAP (DRS-III) program, and DU-DST for PURSE grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Enna Dogra Gupta
    • 1
  • Manendra Pachauri
    • 2
  • Prahlad Chandra Ghosh
    • 2
  • Manchikatla Venkat Rajam
    • 1
  1. 1.Department of GeneticsUniversity of Delhi South CampusNew DelhiIndia
  2. 2.Department of BiochemistryUniversity of Delhi South CampusNew DelhiIndia

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