Download Prostate cancer stem cells Ongoing Projects 3

PROSTATE
CANCER
STEM
CELLS
ONGOING
PROJECTS
‐
3
GENOMIC
AND
EPIGENETIC
CHANGES
EPIGENETIC
CHANGES
include
structural
changes
in
the
chroma-n
and
chemical
modifica-ons
of
the
DNA.
This
results
in
heritable
changes
in
gene
expression.
Epigene-c
regula-on
is
frequently
altered
in
cancer
and
acts
in
combina-on
with
gene-c
changes
during
cancer
ini-a-on
and
progression.
Normal
Tissue
Low
density
CpG
sites
Using
Pyrosequencing
technology,
we
can
study
the
dysregula-on
of
DNA
methyla-on
in
prostate
cancer
stem
cells.
This
will
lead
to
a
beHer
understanding
of
the
mechanisms
that
lead
to
cancer
stem
cell
forma-on
and
to
development
of
new
therapies.
CpG
island
Expressed
Gene
Cancer
Silenced
Gene
Hypomethyla8on
of
low
density
CpG
sites:
• 
genomic
instability
• 
ac-va-on
of
oncogenes
• 
loss
of
imprin-ng
PYROSEQUENCER
Hypermethyla8on
of
CpG
islands:
• 
downregula-on
of
tumor
suppressor
genes
Methylated
CpG
site
Unmethylated
CpG
site
The
best‐known
epigene-c
marker
is
DNA
methyla-on,
which
occurs
primarily
at
CpG
dinucleo-des.
Its
dysregula-on
in
cancer
usually
leads
to
genomic
instability,
ac-va-on
of
oncogenes
and
inhibi-on
of
tumour
suppressor
genes.
Normal
Fusion
GENOMIC
CHANGES
include
dele-ons,
inser-ons
and
transloca-ons
of
DNA
as
well
as
fusion
of
different
DNA
sequences.
This
can
lead
to
genes
being
gained
or
lost
or
being
under
the
control
of
the
wrong
elements.
Increased
expression
of
oncogenes
or
decreseed
expression
of
tumour
suppressor
genes
can
lead
to
cancer.
We
use
a
method
called
FISH
(fluorescent
in
situ
hybridisa-on)
to
detect
these
fusions
by
using
fluorescent
probes
that
are
specific
to
par-cular
DNA
sequences.
DNA
DAMAGE
RESPONSE
We
wish
to
measure
the
response
of
prostate
cancer
stem
cells
to
cancer
therapies
including
radia-on
and
chemotherapy.
Ini-al
therapies
for
prostate
can
be
successful
at
shrinking
the
tumour.
However,
in
some
cases
a
secondary
tumour
emerges
and
this
is
typically
resistant
to
therapy.
We
hypothesise
that
prostate
cancer
stem
cells
are
resistant
to
radia-on
and
chemotherapy
and
are
responsible
for
secondary
tumours.
HOW
TO
MEASURE
DNA
DAMAGE?
DNA
damaging
agents:
‐
Radia-on
‐ 
Chemotherapeu-c
drugs
DNA
break
Cell
detects
DNA
break
using
proteins
that
go
to
site
of
break.
These
proteins
either
signal
repair
or
if
damage
is
too
great,
the
cell
will
commit
suicide.
Protein
is
visualised
by
immunofluorescence
as
dots
in
the
nucleus
of
the
cell
indica-ng
sites
of
DNA
damage.
By
coun-ng
the
cells
with
these
foci
we
can
quan-fy
damage.
Blue
dye
shows
DNA.