2. BACKGROUND
More than 200,000 cases of
breast cancer per year
Leading cause of death in breast
cancers: metastasis
Not many studies related to
glycocalyx
Cancer Metastasis
Nature Reviews
3. EXTRACELLULAR VESICLES (EV)
Extracellular vesicles: particles secreted by cells
Microvesicles, MVs (~200nm)
Can transform healthy cells to cancer-like cells
Prepare premetastatic niches for metastatic
cells
Help transport protein and nucleic acid to
target cells in tumor and body
Exosomes (~100nm)
Antonyak, 2011
4. GOAL
Energetically unfavorable for cell
membrane to bend and release MVs
Glycocalyx could help this process
Studies show that galectin-3, a member
of the galectin family, is enriched in
MVs (Batista, 2011)
Dumic, Dabelic, Flogel, 2006
5. PURPOSE
Hypothesis: MV shedding can be induced in
non-cancerous cell lines that do not express
large quantities of Gal3
1. Introduce Gal3 to non-cancerous cell
lines that do not express Gal3
2. Determine the concentration of MV
shedding in non-cancerous cell lines
transformed to express Gal3
MCF10A Cells
www.ATCC.org
6. METHODS
Experimental groups:
Used MCF10A cells as control
Epithelial cells that do not naturally shed MVs
Introduced 2μM Gal3 in samples
Used same cells but with Gal3 production gene inserted into
DNA
Analyzed by Malvern NS300 Nanosight
Brownian motion
9. GAL3 INDUCES AN INCREASE IN EV
SHEDDING
MCF10A rtTA MCF10A rtTA Gal3 MCF10A rtTA with
2uM Gal3
Total Particle
concentration,
particles/cell
(100nm particles
and above) 5.47x10^7 5.42x10^7 13.86x10^7
Total Particle
Concentration,
particles/cell
(200nm particles
and above) 0.418x10^7 0.403x10^6 2.19x10^7
10. GAL3 INDUCES A SMALL INCREASE
IN EVSHEDDING
MCF10A rtTA MCF10A rtTA
Gal3
MCF10A rtTA
with 2uM Gal3
Total Particle
concentration,
particles/cell
(100nm particles
and above) 5.47x10^7 5.42x10^7 13.86x10^7
Total Particle
Concentration,
particles/cell
(200nm particles
and above) 0.418x10^7 0.403x10^6 2.19x10^7
11. CONCLUSION
No clear evidence that Gal3 affects MVs (200nm and up)
However, 2 fold increase in extracellular vesicles
E.g. exosomes (100nm and up)
Could potentially play a role in cancer metastasis
12. FUTURE DIRECTIONS
Create new cell line that knocks out Gal3 production
Measure particle concentration of cells whose galectin ligands are
modified
Ligands: cell surface sugars
Preliminary data from changed Gal3 ligand shows a decrease in overall particle
concentration
I am not sure if the videos work here; could you check?
I will add a chart with the average total particle concentration of each type of sample here. There will be two rows: only considering particles100nm and up, and only considering particles 200nm and up.
