About OMICS Group

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About OMICS Group

• OMICS Group is an amalgamation of Open
  Access publications and worldwide international
  science conferences and events. Established in
  the year 2007 with the sole aim of making the
  information on Sciences and technology Open
  Access, OMICS Group publishes 500 online open
  access scholarly journals in all aspects of
  Science, Engineering, Management and Technology
  journals. OMICS Group has been instrumental in
  taking the knowledge on Science technology to
  the doorsteps of ordinary men and women. Research
  Scholars, Students, Libraries, Educational
  Institutions, Research centers and the industry
  are main stakeholders that benefitted greatly
  from this knowledge dissemination. OMICS Group
  also organizes 500 International
  conferences annually across the globe, where
  knowledge transfer takes place through debates,
  round table discussions, poster presentations,
  workshops, symposia and exhibitions.

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About OMICS International Conferences

• OMICS International is a pioneer and leading
  science event organizer, which publishes around
  500 open access journals and conducts over 500
  Medical, Clinical, Engineering, Life Sciences,
  Pharmacology scientific conferences all over the
  globe annually with the support of more than 1000
  scientific associations and 30,000 editorial
  board members and 3.5 million followers to its
  credit.
• OMICS Group has organized 500 conferences,
  workshops and national symposiums across the
  major cities including San Francisco, Las Vegas,
  San Antonio, Omaha, Orlando, Raleigh, Santa
  Clara, Chicago, Philadelphia, Baltimore, United
  Kingdom, Valencia, Dubai, Beijing, Hyderabad,
  Bengaluru and Mumbai.
• .

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SLUG and SOX9 Cooperatively Regulate Tumor
Initiating Niche Factors in Breast Cancer
Babak Behnam, MD, PhD Department of Genetics and
Molecular Biology, Iran University of Medical
Sciences behnam.b_at_iums.ac.ir
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Introduction

• Metastasis is the major cause of cancer mortality
• Metastasis is a multistage process
• Successful initiation of metastatic growth,
  metastatic colonization, accomplished only by a
  minority of cancer cells that reach distant sites

Gupta Massague 2006, and Valastyan Weinberg
2011
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Introduction

• A small population of cancer stem cells is
  critical for metastatic colonization
• 1889 Stephan Pagets Seed and Soil hypothesis,
  todays Metastatic Tropism
• - Seed Cancer Stem Cells (CSCs)
• - Soil Metastatic niche

Valastyan Weinberg 2011, and Fidler I.J. 2003
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Cancer Stem Cells

• CSCs Minority proportion of tumor mass
• Self-renewal
• Tumorigenicity
• Multilineage differentiation
• Migration
• Invasiveness
• Apoptosis resistance

Jordan C.T. et al. 2006
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CSCs and Metastatic Colonization
Fazilaty, H. et al., Tumor Biol 2013
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EMT, CSCs and Metastasis

• Epithelial to Mesenchymal Transition
• Embryonic morphogenesis
• In adult
• Wound healing
• Most pivotal escape mechanism of tumor cells
• Migration
• Invasiveness
• Apoptosis resistance
• Expression of Extracellular matrix (ECM)
  components

Thiery J.P. et al. 2009
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EMT Inducers

• EMT transcription factors (EMT-TFs)
• SNAIL, SLUG, TWIST1, TWIST2, ZEB-1, ZEB-2, .
• Signaling pathways
• Transforming growth factor-b (TGF-b)
• WNT/b-catenin
• ..

Craene BD Berx G 2013
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SLUG and SOX9 Master TFs
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SLUG and SOX9 Master TFs

• Actions of key transcriptional regulators suffice
  to convert differentiated epithelial cells into
  long-term repopulating epithelial stem cells
  (SCs)
• Autoregulatory program activated by Slug and Sox9
  is involved in inducing and sustaining the SC
  state
• Adult SCs/CSCs, similar to ES cells, maintain
  their SC state via master regulator-mediated
  autoregulatory networks

Guo et al., Cell 2012
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Survival Rate

• Patients with primary tumors expressing high
  levels of both SLUG and SOX9 had a significantly
  lower overall survival rate than the rest of
  patients

Guo et al., Cell 2012
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Metastatic Niche

• Fertile microenvironment
• Supports stem cell maintenance and manages cell
  function and proliferation
• Cells, vascular networks, soluble factors,
  nutrients, metabolic components and extracellular
  matrix (ECM)
• VEGF, MMPs, LOX

Psaila B. Lyden D. 2009, and Hanahan D.
Weinberg R.A. 2011
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ECM and Metastasis

• Regulatory signals
• Sophisticated organization
• POSTN and TNC, as metastatic niche components for
  CSCs

Malanchi I. et al. 2012, Oskarsson T. et al. 2011
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Periostin (POSTN)
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Tenascin C (TNC )
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Link Between CSCs and Metastatic Niche
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Fazilaty, H. et al., Tumor Biol 2013
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Neovascular tips comprise micrometastatic niches
enriched for POSTN and TGF-b1
Ghajar et al. Nature Cell Biology 2013
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Fazilaty and Behnam, Cell Biology International
2014
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Hypothesis
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Methodology

• Cell culture
• MDA-MB-231 cell line
• MCF7 cell line
• Lentiviral vectors and transduction containing
  SLUG and SOX9 genes their knock down
• Quantitative Real-time PCR
• Check the expression of SLUG, SOX9, TNC and POSTN
  before and after transduction and knock down

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Lentiviral constructs
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Lentiviral constructs
pWPXL-SOX9
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• Results

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• Overexpression of SLUG and SOX9 upregulates POSTN
  and TNC

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MCF7 versus MDA-MB-231
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Quantitative gene expression analysis
P-Value 0.0001 , P-Value 0.001 ,
P-Value 0.01
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Knock-down of SLUG and SOX9 downrgulates POSTN
and TNC
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Sh-RNA Construct
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Quantitative gene expression analysis
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• Discussion

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Coexpression of SLUG-SOX9 and POSTN

• SLUG and SOX9 expression may have synergistic
  effects
• SLUG and SOX9 coexpression is required for POSTN
  and TNC upmost expression
• POSTN is likely one of the most important factors
  regulated by SLUG and SOX9

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Acknowledgment

• Hassan Fazilaty, PhD student
• IUMS Research deputy Grant No. 17856

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References

1. Gupta GP and Massague J. Cancer metastasis
   building a framework. Cell 2006 127(4)679-95.
2. Valastyan S and Weinberg RA. Tumor Metastasis
   Molecular Insights and Evolving Paradigms. Cell
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3. Fidler I J. The pathogenesis of cancer
   metastasis the seed and soil hypothesis
   revisited. Nature Reviews Cancer2003 3(6)453-8.
4. Jordan CT, Guzman ML, and Noble M. Cancer Stem
   Cells. N Engl J Med 2006 355(12)1253-61.
5. Thiery JP, Acloque H, Huang RY, Nieto MA.
   Epithelial-mesenchymal transitions in development 
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6. Craene BD and Berx G. Regulatory networks
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7. Takahashi K, Tanabe K, Ohnuki M, Narita M,
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8. Yang J, Mani SA, Donaher JL, Ramaswamy S,
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9. Lu X, Kang Y. Hypoxia apnd Hypoxia-Inducible
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10. Nieto MA The snail superfamily of zinc-finger tran
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11. Guo W, Keckesova Z, Donaher JL, Shibue T,
    Tischler V, Reinhardt F, et al. Slug and Sox9
    Cooperatively Determine the Mammary Stem Cell
    State. Cell 2012 148(5)1015-28.
12. Psaila B, and Lyden D. The metastatic niche
    adapting the foreign soil. Nat Rev Cancer 2009
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13. Hanahan D, and Weinberg RA. Hallmarks of Cancer
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14. Malanchi I, Santamaria-Martinez A, Susanto E,
    Peng H, Lehr HA, et al. Interactions between
    cancer stem cells and their niche govern
    metastatic colonization. Nature 2012 481 8589.
15. Oskarsson T, Acharyya S, Zhang XH, Vanharanta S,
    Tavazoie SF, Morris PG, et al. Breast cancer
    cells produce tenascin C as a metastatic niche
    component to colonize the lungs. Nat Med 2011
    17(7)867-74.
16. Oskarsson T and Massague J. Extracellular matrix
    players in metastatic niches. EMBO J 2012
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17. Serrano I, McDonald PC, Lock FE, Dedhar S. Role
    of the integrin-linked kinase (ILK)/Rictor
    complex in TGFß-1-induced epithelial-mesenchymal
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18. Ghajar CM, et al. (2013) The perivascular niche
    regulates breast tumour dormancy. Nat Cell Biol
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19. Fazilaty H, Gardaneh M, Bahrami T, Salmaninejad
    A, Behnam B (2013) Crosstalk between breast
    cancer stem cells and metastatic niche emerging
    molecular metastasis pathway? Tumour biology the
    journal of the International Society for. Oncodev
    Biol Med 34 201930.

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