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Analysis of eHsp90 Regulation of Secreted Exosomes from Cancer Cells
Juan Felipe ToscanoMcCready Lab
Department of Natural Science, Assumption College
Samarasinghe, 2013
• Under stress, cancer cells release a high amount of micro-vesicles named exosomes
• Exosomes have a high amount of proteins that could potentially enhance cancer invasiveness and metastasis
• Exosomes could be a way for cells to communicate and exchange genetic information outside the cell
• One of the most abundant proteins identified inside the exosomes was the Heat shock protein 90 (Hsp90)
• Hsp90 is an important intracellular protein that activates many proteins
• eHsp90 is secreted from tumor cells via exosomes
• It activates pro-invasive proteins that contribute to cellular aggressiveness and motility
McCready et al. 2010
Hypothesis
We hypothesize that eHsp90 regulates exosome secretion from cancer cells
MDA-MB-231
Media
Depleted media
Determine protein concentration
Western Blot
6050403020
80110160260
15
Contr
ol 1 H
sp90
Contr
ol 2 H
sp90
Hsp90control Bands
Western blot
ExosomesDepleted
of exosomes
Hsp90
0
5000
10000
15000
20000
25000
Exosome Media
Arbi
trar
y un
its
25
20
15
10
5
0
Hsp90 concentration
MDA-MB-231
100nM STA-12-7191Untreated
DMSO 10nM STA-12-7191
Depleted media
Determine protein concentration
Western Blot
Depleted media
Depleted media
Depleted media
6050403020
80110160260
15
Contr
ol 1 H
sp90
Contr
ol 2 H
sp90
Hsp90control Bands
untreated DMSO10nM STA-12-7191
100nM STA-12-7191
Hsp90
Depleted of exosomes
Western blot from Drug treatment
Hsp90 concentration in media depleted of exosomes
0
10
20
30
40
50
Untreated DMSO 10nM STA-7191 100nM STA-7191
Arbi
trar
y un
its
6050403020
80110160260
15
Contr
ol 1 H
sp90
Contr
ol 2 H
sp90
Hsp90control Bands
Untreated DMSO
10nM STA-12-7191
100nM STA-12-7191
Western blot from Drug treatment
Hsp90
Exosomes
Hsp90 concentration in Exosomes isolated
0
10
20
30
40
50
Untreated DMSO 10nM STA-7191 100nM STA-7191
Arbi
trar
y un
its
Inhibiting eHsp90
• eHsp90 levels remain high in exosome fraction
• eHsp90 levels decrease in media depleted of exosomes
Conclusions• Data suggest that eHsp90 may be important
for protein release from exosomes.
Untreated Cells
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Treated cells
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Acknowledgements
• I want to thank Professor Jessica McCready for giving me the opportunity to work in her lab.
• I would also like to thank the Jay Lab members at Tufts University for giving me the opportunity to continue this research that I started with them back in 2014