Can Expression of VACM-1
Enhance Effects of Thalidomide
on Endothelial Cell Growth?
Drake Harper
Zeeland East High School
John Pelton
Maria Burnatowska-Hledin
Hope College
VACM-1
(Vasopressin-Activated Ca2+-mobilizing receptor)
• Isolated and studied by Hledin’s Lab
• Inhibits Cell growth by regulating cAMP and MAPK
phosphorylation, and p53 concentration, signaling
pathways involved in cell growth that lead to cancer
• In vivo VACM-1 is expressed in endothelial cells (cells in
vasculature)
• Mutation in VACM-1 sequence (where one amino acid is
changed) creates a dominant negative phenotype
(reverse the effect) ie. Cell growth increases
Thalidomide
• Originally created to help pregnant women with “morning
sickness”
• Caused Birth Defects (Physical deformities on limbs and
organs), Later banned
Reason for Study:
Inhibits Angiogenesis
(the growth of blood vessels)
Cellular targets for Thalidomide’s effects are unknown
Hypothesis
In normal endothelial cells expressing VACM1 protein, thalidomide will decrease cell
growth by inducing VACM-1 protein
expression.
In cells expressing a dominant negative
VACM-1 (VACM-S730A), Thalidomide will
increase the mutated VACM-1 but will not
inhibit growth.
Uses on cancer
• VACM-1 stops cell growth, therefore, if
used on cancer cells, it should stop the
growth of cancerous tumors on cells.
Methods
• Cell Culture
• Western Blot
• Wound Healing Assay
• Immunostaining
Western Blot
GAPDH
Data
Control
90
10 μg/ml
80
50 μg/ml
70
60
50
40
30
20
10
Percent Growth in RAMEC -S730 for three treatments of
Thalidomide
0
T=12
T=18
T=24
Control
120
Time (hours)
10 μg/ml
50 μg/ml
100
Percent growth
Percent Growth
Percent Growth of RAMEC CMV Cells with
100
Three Treatments of Thalidomide
80
60
40
20
0
T=12
T=18
Time (hours)
T=24
Summary
• Thalidomide increases VACM-1 protein
concentration which results in decreased
growth.
• When VACM-1 is mutated, it is unable to
inhibit cell growth
Conclusion
Anticancer effect of Thalidomide may
involve induction of VACM-1
Acknowledgements
Dr. Hledin
John Pelton
Dr. Hledin’s lab group
REACH
Hope College