SPEAKER:

ADVISOR:

Received: January 1, 2017,Accepted: March 9, 2017, Published: March 23, 2017

https://doi.org/10.1371/journal.
 l
Belgium has the highest rate of
2nd most common cancer
breast cancer followed by Denmark
overall
and France

Breast cancer

5th most common cancer in

women
 Not bearing children and Progesterone and estrogen
first pregnancy over the
age of 30

Exposure of ionizing Oral contraceptives

radiation
Risk factors

Late natural Hormone therapy

menopause

Early
pubertynarche
 TYPES OF BREAST CANCER

Hormone receptors positive

breast cancer
(Endocrine therapy)

Clinical subtypes

Triple negative breast cancer

ESTROGEN RECEPTOR POSITIVE BREAST CANCER
 CHEMOTHERAPIC DRUGS FOR
ADVANCED BREAST CANCER

Taxanes -paclitaxel (Taxol)

Anthracyclines -
Doxorubicin(Adriamycin), and
Epirubicin

Platinum agents- cisplatin,

carboplatin
 What is Multi Drug Resistance(MDR)?
The cancer cells which have the ability to develop resistance
simultaneously to many different antineoplastic agents is called
Pleotropic resistance or Multi Drug Resistance.
ABC
Epigenetic Apoptotic
transporter
regulation induction
family

DNA repair
Autophagy
and MDR
induction
damage

Hypoxia Cancer
miRNAs
induction stem cells
 SLUG( SNAI2)

TWIST SNAIL

SLUG
 Matrix Metalloproteinase 1(MMP1)
(Interstitial /Fibroblast Collagenase)

Degrades Extra Cleaves cell Releases apoptotic Activates chemokine

Cellular Matrix surface receptors ligands cytokine inactivation.

Differentiation Migration

Host Proliferation
MMP1
defense

Apoptosis
 PREVIOUS STUDIES
NF-kappaB contributes to MMP1 expression in breast cancer
spheroids causing paracrine PAR1 activation and disintegrations
in the lymph endothelial barrier in vitro.2015; pmid:26513020

High MMP-1 mRNA expression is a risk factor for disease-free

and overall survivals in patients with invasive breast carcinoma.
2008. pmid:17663001

The overexpression membrane type 1 matrix

metalloproteinase is associated with the progression and
prognosis in breast cancer. 2015. pmid:25755834

Overexpression of extracellular matrix metalloproteinase

inducer in multidrug resistant cancer cells. 2003.
pmid:12692261
Cdc25A regulates matrix metalloprotease 1 through Foxo1
and mediates metastasis of breast cancer cells. 2011
pmid:21670150

Matrix metalloproteinase 1 and circulating tumor cells in early

breast cancer. 2014. pmid:24972610

The role of MMP-1 in breast cancer growth and metastasis

to the brain in a xenograft model. 2012. pmid:23217186
 The functional role of MMP1 in MDR breast cancer
cells and whether other mechanisms are involved in its
upregulation in the cells have not been fully understood
 OBJECTIVES
To investigate if MMP1 is upregulated in MCF-7/ADR cells
compared to the PARENTAL MCF-7 cells

To examine whether high MMP1 expression is associated with

worse survival outcomes in breast cancer patients after systemic
therapy

To check if upregulation of MMP1 results in enhanced MDR in

MCF-7 cells

To check whether slug modulates MMP1 expression in MCF-

7/ADR cells

To investigate whether Slug upregulation leads to aberrant MMP1

expression via direct binding to the MMP1 promoter
 AIM 1
To investigate if MMP1 is upregulated in MCF-7/ADR cells
compared to the PARENTAL MCF-7 cells

MCF-7 : Michigan Cancer Foundation-7

MCF-7/ADR: Adriamycin Resistant cells
Microarray data comparing the dysregulated genes in MCF-7/ADR
and parental MCF-7 cells

MMP1 CDH1

MMP1 is upregulated in MCF-7/ADR cells compared to

parental MCF-7 cells
 AIM 2

To examine whether high MMP1 expression is associated with

worse survival outcomes in breast cancer patients after systemic
therapy

Chemotherapy, Endocrine therapy

High MMP1 associated Overall Survival(OS) and Recurrence
Free Survival(RFS) in all breast cancer patients
High MMP1 associated Overall Survival(OS) and Recurrence
Free Survival(RFS) in ER+ breast cancer patients
High MMP1 associated Any Event (AE) and Metastatic
Relapse(MR) Free Survival in all breast cancer patients
 High MMP1 associated Any Event (AE) and Metastatic
Relapse(MR) Free Survival in ER+ breast cancer patients

High MMP1 expression is associated with worse OS, RFS, AE

free survival and MR free survival in breast cancer patients
after systemic therapy
 AIM 3

To check if upregulation of MMP1 results in enhanced

MDR in MCF-7 cells
Overexpression of MMP1 in MCF-7 cells causing increased
MDR
Knockdown of MMP1 in MCF-7/ADR cells causing
increased sensitivity in drugs
Screening of apoptosis without Adriamycin in cells with
differentially expressed MMP1

Caspase-9 - cysteine-aspartic proteases involved in apoptosis

and cytokine signaling
Apoptosis in overexpressed/ knockdown of MMP1 in
cells without Adriamycin
Screening of apoptosis with Adriamycin in cells with
differentially expressed MMP1
 Apoptosis in overexpressed/ knockdown of MMP1 in
cells with Adriamycin

MMP1 upregulation results in enhanced MDR in MCF-7 cells

 AIM 4

To check whether slug modulates MMP1 expression in

MCF-7/ADR cells
Checking Slug expression which facilitates EMT in MCF-7/ADR
and MCF-7 cells
 Effects of differentially expressed slug on MMP1

Slug mediates the MMP1 expression in MCF-7/ADR cells

 AIM 5

To investigate whether Slug upregulation leads to aberrant

MMP1 expression via direct binding to the MMP1 promoter
Screening of direct binding site of slug to MMP1 promoter

-700 -500
-410 -300
ChIP assay procedure

5’ttttaatgggcaggagatgc3’ FORWARD PRIMER

5’ggatgatgaaaaggctggaa3’ REVERSE PRIMER
 Chromatin Immuno Precipitation assay verifying
direct binding of slug to MMP1 promoter

Slug leads to MMP1 expression by direct binding to MMP1

promoter region
 CONCLUSION

 MCF-7/ADR cells had significantly upregulated MMP1

expression than the parental MCF-7 cells.

 High MMP1 expression is associated with worse survival

outcomes in breast cancer patients after systematic therapy.

 MMP1 knockdown reduced the MDR of MCF-7/ADR cells.

 Slug directly promotes MMP1 transcription, which is a

previously unrecognized mechanism of MMP1 upregulation
in MDR breast cancer cells.
 DISCUSSION
 Only limited studies investigated the involvement of MMP1 in
drug responses of the cancer cells.

 Further investigation on the functional role of MMP1 in drug

responses of the cancer cells led to the findings that, MMP1
expression might be related to EMT of the cancer cells.

 Based on the observation that MCF-7/ADR cells had enhanced

mesenchymal properties, it enforced that Slug expression
substantially increased MMP1 level in MCF-7 cells.

 while knockdown of Slug significantly decreased MMP1 level

in MCF-7/ADR cells

 MMP1 is one of the downstream effectors of Slug in modulating

chemo-sensitivity of breast cancer .