Vol. 2, 1829-1835,
November
Inductively
Clinical
1996
Coupled
Platinum-DNA
Patients
Plasma
Adducts
Receiving
Mass
Spectroscopy
in Peripheral
Cisplatin-
Cancer
Quantitation
Blood
Leukocytes
Research
1829
of
of
or Carboplatin-based
Chemotherapy’
Andrea
Bonetti,2
Flavia
Piero
Pavanel,
Gian
Luigi
Loris
Sperotto,
Department
University
Apostoli,
Marco
Cetto,
of Medical
of Verona,
higher
Zaninelli,
and
Franceschi,
Roberto
Ospedaliera
and
[A. B., M. Z., F. P.,
G. L. C., T. F., L. S.]; Institute of Occupational
Medicine,
of Brescia, 25121 Brescia, Italy [P. A.]; and Institutes of
Pharmacology
[R. L.] and Immunology
[M. C.], University
Verona, 37121 Verona, Italy
copy
and
lated
previously
adducts
by means
ELISA,
and
with
num-based
chemotherapy.
formation
in peripheral
method,
response
and
Platinum
of ICP-MS
beginning
platin
have
clinical
been
were
and
tin
corre-
response
to plati-
adduct
by means
of a new
measured
exposed
to drug
(mean
± SD),
in
level
was
vitro
value
of 23.4
nine healthy
administration
either
in
vitro
before
with
of drug
cis-
to the
in leukocytes
14.33
from
14.71
±
patients
fmol/p.g
volunteers.
In samples
of chemotherapy,
collected
DNA
Pt-DNA
after
adducts
in leukocytes
adduct
levels
from
chemotherapy
tively).
(3.15
exposed
(r
=
patients
0.085
At 24 h, adduct
3.64
±
in
fmol/p.g
and
levels
DNA,
did
vitro
not
the
ranged
treated
with
0.011
at 1 and
in patients
mean
±
SD)
were
To whom
Medical
37121
requests
Oncology,
Verona,
Italy.
for reprints
Azienda
Phone:
should
be addressed,
and
received
dose
intensity,
creatinine
in patients
suggest
with
adduct
a major
determinant
analogue
carboplatin
not
formation.
that ICP-MS
treated
that
hemo-
could
be
al-
cispla-
formation
of
response
the
are
or
antineoplastic
(1 ).However,
for the majority
of these cancers,
exception
of germ
cell tumors,
the proportion
sponding
patients
of a marker
could
is usually
that
would
in
of immunochemical
normal
Previous
eral
blood
tissues
reports
treated
in
identification
to chemotherapy
can
be
adduct
from
levels
patients
with
has been
in periph-
clinical
made
could
by
(6).
receiving
the
in leukocytes
form
adduct
determined
(3-5) or by AAS
chemotherapy
formation
and carboplatin
(2). Pt3-DNA
tumors
the Pt-DNA
collected
cisplaactivity
that the level
be a marker
of
(3, 7-10).
Furthermore,
a correlation
has been
adduct formation
in leukocytes
collected
from
and adduct
thus suggesting
formation
in
chemotherapy
cisplatin
in DNA
and the suggestion
adduct
patients
leukocytes,
significantly
the
response
methods
link
leukocytes
chemosensitivity
reported
between
cisplatin
and
or carboplatin-based
of Pt-DNA
24 h, respec-
therefore,
the
acquation,
adducts
means
of the two drugs,
with
small;
predict
with the
of re-
be useful.
levels
tin-
cisplatin-based
receiving
Received
5/10/96;
revised
8/8/96;
accepted
9/3/96.
‘ This
work
was supported
in part by a grant from
Italiana
per la Ricerca
sul Cancro.
2
correlate
and
is not
After intracellular
intraand interstrand
from 1.91 ± 3.59 fmol4ag
DNA (mean ± SD) at the 1-h time
point to 2.61 ± 3.35 fmol/ig
DNA at 24 h (P > 0.05). Adduct
levels
5FU,
posttreatment
of adducts
carboplatin
cancers
notable
which was not significantly
different
from the
± 19.53
fmol/ig
DNA observed
in leukocytes
from
and
and
Cisplatin
a cisplatin-
collected
incubated
and
agents with demonstrated
activity
in different
types of cancer,
particularly
testicular,
ovarian,
lung, head and neck, and bladder
by means
receiving
or
DNA)
INTRODUCTION
spectroscopy
with clinical
the administration
adduct
fmol/.ag
evident
toxicity.
was to study
leukocytes
chemotherapy,
Pt-DNA
± 3.51
0.071).
=
were
3.01 fmol4ag DNA) to chemotherapy.
group of patients
treated with combi-
in leukocytes
levels
patient.
The
of cisplatin
decrease,
(3.23
SD)
±
the latter
(P
±
In the homogeneous
globin
although
(mean
DNA)
high-dose
significance
levels
responsive
(2.34
nation
DNA),
adduct
adducts
or 1 and 24 h after
in adduct
treated
fmol/ag
receiving
the detection
of 66 patients
of treatment
in patients
(1.18 ± 1.06 fmol4ag
did not reach statistical
patients
± 0.73
lows
Our purpose
or carboplatin-based
those
in patients
(0.57
can be assayed
in peripheral
of atomic
absorption
spectros-
blood
in leukocytes
than
nonresponsive
of
observed
with the extent of leukocyte
adduct
The data presented
here demonstrate
favorable
toxicity.
(Pt)-DNA
higher
between
University
those
linked
inductively
coupled
plasma
mass
and to correlate
adduct
formation
(ICP-MS),
the
high
also
than
dose carboplatin
No differences
ABSTRACT
Platinum-DNA
blood
leukocytes
0.02)
=
carboplatin
difference
Leone
Oncology,
Azienda
37121
Verona,
Italy
(P
with standard
Colombatti,
Tiziano
and
Marta
formation
in in vitro
the possibility
vitro could
help
(1 1, 12). A similar
cisplatin-treated
that the rate of adduct
in predicting
conclusion
the
was
response
reached
to
when
the Associazione
The abbreviations
used are: Pt,
spectroscopy;
ICP-MS,
inductively
3
at Department
of
Ospedaliera
di Verona,
P.le Stefani
39 45 8072351;
Fax: 39 45 8341277.
I,
5FU,
5-fluorouracil;
concentrations
over
DI, dose
time;
CI,
platinum;
coupled
intensity;
confidence
AAS,
plasma
AUC,
atomic
absorption
mass spectroscopy;
area under
the curve
interval.
Downloaded from clincancerres.aacrjournals.org on June 17, 2017. © 1996 American Association for Cancer
Research.
of
1830 Quantitation
the
of Platinum-DNA
DNA
stop
damage
assay
study,
from
treated
there
vivo
and
in a large
in vitro
and
ICP-MS.
source
dynamic
seventeen
and
gave
history,
< 14
(total
upper
X-ray;
count
normal
2.
If
never
Cooperative
patients
have
atinine
had
Cisplatin
single
bolus
was
(1-h
posthydration)
infusion
esophageal,
and
and
mitomycin
tients);
or cyclophosphamide
clophosphamide
cancer patients).
20
mg/rn2
posthydration)
nitrogen
platelet
count
hepatic
less
than
func-
two
on chest
expectancy
Group
times
>3
months;
performance
previously,
status
they
nephrotoxicity
must
(peak
sodium
5FU
7
5
3
Cisplatin-cyclofosphamide-adriamycin
3
Others
4
chemotherapy
(AUC 3-6 mg - min/mL)
(AUC 12-18 mg - min/mL)
non-small
cell lung cancer.
1000
of 100 mg/m2
chloride
with
1000
mg/m2/day
10
6
mg/m2
infusion
on days
in 3%
1-5
sodium
in combination
for 12 weeks
(head
and
cell
single
agent,
chloride
with
with
Standard-dose
as a single
alone
or with
patients),
carboplatin
(300-400
mg/rn2)
was admindose in 5% dextrose
in water over 1 h, either
cyclophosphamide
etoposide
High-dose
1 h on days
1-4)
was
g/m2 (with mesna
over 4 consecutive
Planned
Hryniuk
age
given
weekly
mg/rn2
and received
and
in combination
as a uroprotector)
days.
Bush
(ovarian
planned
was
determined
received
DI.
The
with
and
DIs were
(16);
of the
AUC
600
calculated
DI was
0.00457
x age)
micromolar
X (1
concentration
and
sex
received
The analysis
weight
and
of toxicity
sex
and
carboplatin
carboplatin
according
to Chat-
X weight
X (1
-
expressed
in kilograms,
in
age
in
I if female).
=
of received
performed
col-
lected by venipuncture
or cy-
and
bleo-
of cisplatin
chemotherapy.
(or carboplatin),
Immediately
ered in three
50-ml
after
tubes
10 ml of hystopaque-1077
and
then
supernatant
centrifuged
was
(10
at 700
and washed
removed,
during
and
blood
cycle
was
10 ml of hystopaque-l
ml of blood
X g for
twice
sodium
the end
the first
the collection,
containing
and
collected
be reported
in syringes
containing
heparin
of treatment
and at 1 and 24 h after
the beginning
of infusion
but will
DI was
and
before
cycles
of
as percent-
x sex)]/creatinine
(with
0 if male
=
12
mg/rn2
by the method
the administered
0.314
-
1200
expressed
effectively
by dividing
ifosfamide
etoposide
dose by the carboplatin
clearance,
calculated
elut’s (17) formula:
0.134
X weight + [218
years,
cancer
or in combination
with doxorubicin
SO mg/rn2
and
100 mg/rn2 on days 1-3 (cancer
of unknown
primary).
carboplatin
(300 mg/rn2 in 5% dextrose in water over
as a
pa-
pre-
on days
only
neck,
cancer
100 mg/rn2
here
6 mg/rn2
lung
and etoposide
for the patients
treated
with the cisplatin-5FU
combination.
Blood
Sample Collection.
Thirty
ml of blood were
as a
pre-
or vinblastine
(non-small
(30 units)
for all the cisplatin-based
days
patients);
istered
cre-
500 mg/rn2 and doxorubicin
50 mg/rn2 (ovarian
Patients
with testicular
cancer received
cispla(I-h
28
1-S.
diagno-
urea
adequate
5 consecutive
C 10 mg/rn2
blood
of cardiomegaly
with:
cancer
were
(15).
at the dose
in 3%
over
and
confirmed
p.M.
of
neurotoxicity.
administered
colon
beginning
cells/pA,
cisplatin-induced
infusion
the
and toxicity
criteria
cisplatin
in combination
continuous
the
life
or disabling
i.M)
before
Oncology
received
suffered
>195
biochem-
and
hearing:
based
results
WHO
no evidence
Patient
was
a routine
transaminases
p.M;
of normal);
31, 1995.
Response
g/dl,
in this
and
>3,000
>9
22 to 72 years
count,
of < 134
hemoglobin
clinically
3
9
Cisplatin-etoposide-bleomycin
Cisplatin-cyclofosphamide
mycin
Forty-nine
staging
a pathologically
leukocyte
and an Eastern
cell
to standard
<26
from
1 . Clinical
patients
included
bilirubin
6
Cisplatin-vinblastine-mitomycin
Low-dose
High-dose
a NSCLC,
wide
to participate
examination,
all
creatinine
limit
in age
and December
blood
serum
> l00,000/i.l,
cancer
Carboplatin-based
de-
as platinum;
Schedule.
consent
of chemotherapy.
criteria
riM,
ranging
physical
according
of cancer;
throughput.
sample
determi-
excellent
high
in Table
for
cycles
element
such
1, 1994,
procedures
of
determinations;
informed
a complete
Entry
for trace
ion
method
23
10
8
7
Breast cancer
Others
Cisplatin-based
chemotherapy
Cisplatin-5FU
spectrom-
ICP-MS
22-72
Testicular
coupled
is
54
Range
Esophageal
cancer-head
and neck cancer
Lung cancer (NSCLC)
Ovarian cancer
Cancer of unknown
primary
as
an interface
Median
METHODS
are listed
evaluated
The
for elements
women
54)
used
of patients
17
the
for
millibars,
and Treatment
January
profile,
plasma)
No.
Diagnosis
of an
spectrometer
l0
components.
AND
three
than
advantages
Selection
characteristics
tin
two
range;
on a complete
mass
multielement
MATERIALS
tion
better
in particular
between
an argon
66)
=
49
in
is comprised
the mass
linear
after
instrumentation
quadrupole
for
of elements
and
limits,
imaging
and
technique
amounts
pressure
the
age,
(n
Female
Age (year)
was used:
at atmospheric
tection
ical
a
adducts
analytical
(normally
has several
and
of Pt-DNA
inductively
simultaneous
Patient
of patients
Male
in
formation
the
nation:
study
The
a vacuum
measurement
(median
with
formation
Because
between
men
collected
quantification.
works
eter requires
Characteristics
Sex
the hypothesis
adduct
ultra-trace
plasma
and
and
placed
in
in vitro
adduct
leukocyte
and
(14).
coupled
detection
leukocyte
is a powerful
samples
plasma
Table I
PCR-
measured
incubated
of patients
cohort
between
of trace
inductively
were
in leukocytes
and
for the assay
ICP-MS
biological
and
between
and
method
determination
ion
by the
toxicity.
A new
sis
patients
is a correlation
response
adducts
of drugs,
to confirm
that
determined
Pt-DNA
the administration
cisplatin
was
Characteristic
present
leukocytes
an
in leukocytes
(13).
In the
before
Adducts
30 mm.
in PBS
the pellet
solution
was
in each
Leukocytes
of
lay119
tube),
were
(pH 7.4);
the
frozen
at - 20#{176}C
Downloaded from clincancerres.aacrjournals.org on June 17, 2017. © 1996 American Association for Cancer
Research.
Clinical
ICP-MS
AAS
Standard
curves
by
ICP-MS
for platinum
and
0.05
.
de-
by
0.06
a
70
termination
55
AAS.
The lowest detection
limit was 0. 1 .i.g/
liter for ICP-MS
and 10 igIliter
for
AAS.
0.04
!;
40
0
25
0.03
<
0.02
10
0.01
0
-5
-0.1
0.1
0.3
0.5
0.7
0.9
1.1
1.3
1.5
0
100
50
Pt ig/L
and 1 mM EDTA
Louis,
MO) were
(pH 8.0)]. SDS and a protease
added to lyse leukocytes
and
proteins.
were
Proteins
in presence
with
the
in TE buffer
100
(Sigma,
St.
to degradate
z
phenol-chloroform
DNA was precipitated
by absolute
The DNA was then rinsed with 70%
of salt.
and resuspended
extracted
[10 mrvi Tris-HC1
and
ethanol
ethanol
I
S
10
.:#{149}
#{149}
I
#{149}#{149}
0
#{149}Is
::
.:
1 msi EDTA
.
.S.
S.
(pH
8.0)].
DNA
concentration
was measured
by A260.
The
DNA
study
Vitro
the
in vitro
collected
apy
formation
containing
incubation
cisplatin
period,
bated,
treated
then
collected
following
from
in
nine
were
analyzed
5000
Perkin-Elmer
for their
Samples.
adducts,
after
by
H2PtCI3H2O
Edison,
NJ).
Total
dilution
Measurements
in 2 ml of a
made
were
a stock
of
Statistical
icance
with
I compares
spectroscopy.
Analysis.
of the differences
Student’s
t
standard
lowest
limit
of
Pt; the coefficients
and from 5 to 10%
for ICP-MS
and
the
Unless
stated
otherwise,
experimental
the signif-
data
was
of chemotherapy
tered over the course of this study.
cisplatin-based
cycles;
10 patients
based
cycles,
given
at the standard
Fifty
patients
received
target
were
AUC
adminis-
received
40
156
in three
with
mg-min/
66) treated
=
the
a separate
60).
lowest
leukocyte
, median.
nine high-dose
target
AUC
regimens
of 12-18
data
for Pt-DNA
in 13 samples
patients
of chemotherapy.
1 1 samples
of
samples
levels
Pt-DNA
of the
of the second
incidence
The
(mean
chemotherapy
± SD; 95%
chemotherapy
were
higher,
treated
cycle
in
Pt-DNA
DNA
Fig.
first
adduct
number
of planned
and 41 collected
at
time
earlier
point
below
the
time
(22%),
detection
levels
were
undetectable
at both
treated
with
carboplatin
and
time
in three
cisplatin.
after
cytes
the
overall
Leukocyte
carboplatin-
of 3-6
and
of the method.
treated
(mean
cycles
after
undetectable
a 24%
limit
fmol/p.g
RESULTS
received
at the
the individual
point
(27%)
giving
tested
test.
hundred-five
(n
represents
line
in 101 samples
(77% of the total
60 collected
at the 24-h time point
were
point
after
Two
from patients
Dotted
S represents
41, samples at 24 h
at 1 h
2 shows
1-h time
adducts
points
between
in leukocytes
chemotherapy.
Each data point
mg-mm/mb.
Fig.
curves
24 hours
ml; the remaining
six patients
containing
carboplatin
given
determined
samples):
The
adducts
(samples
value
detection
of the assay was 10 pg of elemental
of variation
ranged
from 1 to 3% (intraassay)
(interassay).
Fig.
atomic
absorption
Pt-DNA
platinum-based
of detection.
as the integrated
DNA).
Pt/i.g
Fig. 2
Inc.,
crogram
(fmol
1 hour
Industries,
of 3 mm. Pt-DNA
adduct
of elemental
Pt per mi-
DNA
Sampling
an Elan
curves
of
(SPEX
.............
by a quan-
of ion number
in an analysis
time
levels
were calculated
as femtomoles
ofcellular
...........
samples
using
samples
dilutions
acid
were
DNA
Standard
serial
hydrochloric
S
S
0.1
with
limit
leukocytes
determined
(Sigma).
aqueous
10%
Pt-DNA
in
by ICP-MS
of the DNA
S
S
sample
Pt was
in H20
using
in
evaluated
the
iso-
volunteers.
Pt content
S
#{149}#{149}.
leukocytes
above.
Adducts.
#{149}.I
S..
To
15 i.g/ml.
After
and DNA was
described
also
Pt-DNA
Sciex.
solution
generated
was
healthy
of
method
Triton
the steps
vitro
Determination
0.1%
Pt-DNA
at a concentration
of
the cells were washed,
formation
titative
of
of Blood
#{149}#{149}#{149}
from 45 patients
before
the beginning
of chemotherincubated
in vitro
at 37#{176}C
for 2 h in RPM!
1640
were
adduct
Treatment
Cisplatin
:1
. .
0
isolated
from each sample
averaged
690 ± 527 p.g/ml (mean
±
SD); the grade of purification
of DNA was on average
98%.
In
150
Pt tg/L
until analysis.
After thawing
the sample,
the pellet was resuspended
in a DNA buffer [TNE:
10 mri Tris-HC1,
100 mi NaC1,
method, and purified
Research
0.07
85
Fig. 1
Cancer
adduct
± SD;
time
CI
although
3 depicts
in vitro
(P
levels
95%
point
0.05).
with
±
3.35
in samples
With
not statistically
individual
from
both
adduct
No
1-h
DNA
collected
drugs,
± 3.59
at the
fmol/p.g
at 24 h
plasma
different,
Pt-DNA
cisplatin.
1.91
0.81-3.01)
=
to 2.61
1.76-3.46)
>
ranged
CI
levels
at 24 h.
levels
significant
in leukodifferences
Downloaded from clincancerres.aacrjournals.org on June 17, 2017. © 1996 American Association for Cancer
Research.
1831
1832 Quantitation
of Platinum-DNA
Adducts
Table
:
Association
2
b etween
adduct
chemotherapy
levels
at 24 h and
re sponse
to
S
fmol Pt4i.g DNA
(mean ± SD)
S
3.23
2.34
.
S
S
#{149}5
.
S
Response
± 3.51
± 3.01
Complete
Stabilization
partial
(n =
and progression
and
20)
(a
28)
=
#{149}#{149}#{149}SS
z
S
S:
10
..
CI
0,
IS
0
2. 1-4.2)
=
observed
(mean
.::S.
± SD
S#{149}
S
increase
Patients
adducts
( 15 p.g/ml).
cisplatin
sample
(patients
Healthy
in
leukocytes
after
volunteers
vitro
Each data point #{149}
represents
45, healthy
volunteers
9).
Pt-DNA
adduct
was
The
adduct
association
levels
analysis
(n
6.0
were
excluded
six
4).
showed
4.0
3.0
peak
0
2.0
J
I
Carboplatin
high
Cispiatin
100mg/rn’
dose
Mean leukocyte
Pt-DNA
adducts in patients treated with cisplain at the dose of 100 mg/m2 (n
46), standard dose carboplatin
(n
8), or high-dose
carboplatin
(n = 6). Sampling
was at 24 h postinfusion.
Bars, SD.
4
patients
(mean
±
SD
10.03-18.63)
=
23.4
Pt-DNA
±
19.53
adduct
=
and
fmol/p.g
14.33
DNA;
24 h (n
0.085
24
=
29)
and
analogue
administered
of carboplatin,
with
14.4
the level
chemotherapy
±
2.3
as
95%
CI
of adduct
levels
a function
(cisplatin
the effectively
mg-min/ml
groups,
respectively).
with cisplatin
(mean
14.71
=
in the
leukocyte
fmol/p.g
volunteers
cisplatin-based
adduct
levels
and 0.01 1, respectively).
Fig. 4 shows Pt-DNA
adduct
h after
between
±
healthy
In patients
receiving
no correlation
between
was
levels
±
7.12-31.94).
chemotherapy,
at 1 h (n =
formation
in leukocytes
of the type
95%
SD
=
(r
collected
of platinum
or carboplatin)
and, in the case
received
AUC (4.6 ± 1 .2 and
standard
and
in the
adduct
homogeneous
group
cisplatin-5FU,
the
0.22-
=
did
the
patients
receiving
to
increased
the
high-dose
The value of adducts
in patients
treated
± SD = 3.15 ± 3.64 fmoLlp.g
DNA; 95%
to chemotherapy
and
in 48 patients.
treated
tumors,
were
15).
level
and
From
with
six
observed
the
high-dose
patients
(42%;
Although
than
the
who
complete
responsive
patients
difference
of treatment
received
DI
of patients
majority
76) had evaluable
adduct
formation.
observed
between
posttreatment
patients
showing
was
disease
not
significant
(hemoglobin
were evaluated
treated
of
(25
whom
adduct
levels
with the combination
of 28; administered
at 24 h. In this
and
subset
of
at 24 h (1 .97 fmol/p.g
DNA)
two groups
with low and high
As shown
in Fig. 5, no
the two groups
in terms
creatinine,
decrease,
for the
hemoglobin
differences
of
were
received
DI,
decrease.
DISCUSSION
study,
blood
times
(Fig.
I). This
of Pt-DNA
of patients
gave a good
sensitive
than
more
focusing
biological
the quantitation
leukocytes
chemotherapy
This method
adducts
there
25) or at
in vitro
cell
response,
carboplatin-based
from
DNA;
(mean
germ
or progression,
In this
CI
CI
2).
ripheral
in
the
response
patients,
the median
adduct
level
was chosen
as a cutoff to separate
I
Carboplatin
standard
dose
in
proportional
investigated
Finally,
the toxicity
creatinine)
and the
cycles,
0.0
levels
not
20 responses
a higher
(Table
found
95%
because
the treatment
was administered
in an
(n = 2) or because
of early
loss
to follow-up
5; partial
stabilization
were
that
carboplatin
this latter difference
0.071).
Furthermore,
=
the six patients
with
Overall,
response,
5.0
Fig.
than
dose
DNA;
although
(P
between
at 24 h was
were inevaluabbe
adjuvant
setting
1.0
fmol/pg
0.02)
=
AUC.
incubation
with
a separate leukocyte
, median.
in
7.0
0
0.73
±
(P
standard
also higher
than the adduct
levels
in the highgroup
(mean
± SD = 1.18 ± 1 .06 fmol/p.g
carboplatin
in
high-dose
carboplatin,
z
with
DNA; 95% CI = 0.21-1.91),
not reach statistical
significance
S
S
Pt-DNA
higher
treated
0.57
=
1 .24), and was
dose carboplatin
S
3
significantly
patients
S
S
Fig.
was
in
was
reproducibility
standard
AAS
is in agreement
or
ICP-MS.
about
100
Pt-DNA
two published
papers
on the use of ICP-MS
for the determination
of Pt in
samples
such as blood (18), urine, and tissue speciThe sensitivity
of the ICP-MS
in 76%
of the
samples.
below
detection
limits
(19).
the
reported
in
the
only in a paper
ELISA
method
37%
by
and was
in detecting
with
in pe-
a cisplatin
performed
of adducts
mens
adducts
receiving
only
of
the
In previous
leukocytes
have
literature;
of the
we
found
by Reed et a!. (3),
and found detectable
The
allowed
the detection
percentage
of samples
different
this
methods
important
who used
Pt-DNA
is rarely
information
a very
adduct
sensitive
levels in
samples.
studies,
been
Pt-DNA
determined
adducts
by
ELISA
in peripheral
blood
(3, 5, 7), AAS
Downloaded from clincancerres.aacrjournals.org on June 17, 2017. © 1996 American Association for Cancer
Research.
Clinical
%
tmo1/L
Cancer
1833
Research
g/dl
100#{149}
90
80
70
60
50
40#{149}
30
20
10
0
Received
DI
Post-treatment
creatinine
Hemoglobin
decrease
Fig. 5 Mean of received DI, posttreatment
creatinine,
and hemoglobin
decrease in patients treated with the combination
(n
12) with Pt-DNA
adducts
1.97 fmol
Pt/p.g DNA; U, patients (n
13) with Pt-DNA
adducts
> 1 .97 fmol Pt/p.g
Table 3
Published
First
studies
ev aluating
platinum-DNA
addu cts formation
in leukocyte
s and
tumor
cisplatin-5FU.
Bars,
to pla tinum-based
response
chemotherapy
Correlation
author
(reference)
Tumor
Reed et al. (3)
Reed et al. (7)
Parker et al. (8)
Reed et al. (9)
Schellens
et al. (10)
Motzer et al. (22)
Gupta-Burt
type
of assay
Adducts/Response
Testis
and ovary
Ovary
Different
types
45
55
21
ELISA
ELISA
AAS
Positive
Positive
Positive
Different
Different
49
45
AAS
AAS
Positive
Positive
36
ELISA
Negative
AAS
Negative
ELISA
Positive”
Negative
types
types
Testis
et al. (23)
Method
No. of patients
Ovary
and
breast
52
AAS
Only
a
(8-10,
in patients
with ovarian
20, 21), or both
by AAS
are usually
(22,
such
as
the
doses
of cisplatin,
tion,
it is impossible
already
published
onstrated
that
the blood
collection
cancer.
23). Mean
in the range
to two orders
of magnitude
However,
because
the patients
variables,
type
of
24 h in patients
who
AUC.
Although
larger
group
formation
help
adjust
comparison
done
followed
with
in this
the kinetic
study)
of reaction
did not parallel
this
observation
of patients,
of Pt-DNA
the dose
on plasma
AUC.
A correlation
carboplatin
leads
based
it might
adducts
to
patients,
the difference
needs
the utility
in the
high-dose
interactions
lection
tients
relation
Pt-DNA
adduct
formation
to several
gators
(Table
ent degrees
different
setting
to
than
adduct
levels
has been
reported
in leuko-
such
as the proce-
of sensitivity
drugs
used
in tumors.
formation
properties
either
autopsy
studies
the question
shared
by
to
as
with ovarian
by previous
drugs
with
cancer
investi-
conceivably
and
factor
or to the
does
this
(6, 25, 26),
of whether
tumors
and
response
(3, 7-10),
to the confounding
simply
samples
and
pa-
The cor-
could be due to the difare likely to show differ-
combinations,
Regarding
in tumor
between
as positive
to the platinum
in the
evident
in leukocytes
of response),
adduct formation
from
were
to chemotherapy.
3). These
discrepancies
of tumors
studied
(which
formation
adduct
levels
only to patients
by ELISA
(23)
leukocyte
only
factors,
in the
This
incubation
of whole blood instead
the different
assay
method,
or the se-
in adduct
thresholds
the other
in a
is not confirmed
group
of patients.
(i.e.,
and nonresponsive
between
chemotherapy
that raised
between
be due
differences
responsive
cis-
of studying
colhas
of the patients.
No
ferent types
in the target
rather
could
dure of in vitro treatment
of isolated
leukocytes),
this
at
the differ-
to be confirmed
suggest
on drug-DNA
with
previously
(1 1, 12), but
which
involves
a larger
lacking
(22), or as limited
who had adducts
assayed
of carboplatin
compared
discrepancy
after
lower adduct
levels than the
thawing
frozen
whole-blood
In carboplatin-treated
levels
the
dern-
immediately
reported
study,
present
isola-
it has been
incubated
in vitro with cisplatin
and in leukocytes
treated patients,
measured
by an ELISA
method,
lected from
been
the
for DNA
Particularly,
of leukocytes
received
patients.
in adduct
the
one
is slower
than that of cisplatin
(2, 24), and
can explain
the lower level of adducts
observed
platin-treated
ence
that
about
chemotherapy,
followed
a direct
values.
(procedure
known
cytes
determined
DNA,
administered
to make
adduct
values
fmol/p.g
higher
than in the present
study.
studied
differ in terms of several
the determination
of significantly
isolation
of leukocytes
done after
with DNA
observation
adduct
5-500
and the procedures
the isolation
samples
(21).
It is well
E, patients
SD.
DNA.
last
not
parallel
aspect,
in normal
and
fact
it was
data
tissues
these
of
that
adduct
linking
come
studies
there were pharmacogenetic
normal
tissues
of a given
Downloaded from clincancerres.aacrjournals.org on June 17, 2017. © 1996 American Association for Cancer
Research.
1834 Quantitation
of Platinum-DNA
individual.
However,
cepted,
and data
of correlation
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and
rate,
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were
having
either
a high
might
be due
(29,
representation
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as
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action.
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