1:
Neoplasma 2002;49(4):272-7
Deactivation
of P-glycoprotein by a novel compound, oxalyl bis
(N-phenyl)
hydroxamic
acid.
Department
of Environmental Carcinogenesis & Toxicology; Chittaranjan
National
Cancer
Institute,
A
plasma membrane glycoprotein (P-gp) of 170 kd is over-expressed in most of
the
drug
resistant cells. P-gp is encoded in humans by the gene mdrl and is
thought
to
function as a broad substrate ATP-dependent drug efflux pump. P-gp is
also
present
in many types of normal cells. A good number of chemicals inhibit
or
deactivate
P-gp and thus reverse multidrug resistance (MDR). Most of
the
reported
resistance modifying agents (RMAs) are effective in vitro and
have
adverse
effect on the hosts. Hence, the development of nontoxic RMA is
of
immense
importance in the field of cancer chemotherapy. With this end in view,
a
nontoxic
resistance modifying agent, viz., oxalyl bis (N-phenyl) hydroxamic
acid
(OPHA)
has been developed on the basis of the structural commonalities of
the
reported
RMAs. We reported earlier that OPHA reverses doxorubicin resistance
in
vitro
and also reduces glutathione and glutathione S-transferase in a non
P-gp
expressing
cell line. In the present report, the inhibition of P-gp by
the
compound,
OPHA in human cervical cancer cell line, HeLa, has been described
by
western
blotting, study of immunofluorescence and enzyme linked
immunofluorescence
assay (ELISA). The inhibition of P-gp by OPHA is
significantly
higher than that of verapamil. The high IC50 values of
OPHA
against
different cell lines indicate the non toxic nature of the compound.
This
work
underscores the possibility of using the present hydroxamic acid
derivative
as
the nontoxic modulator of the MDR phenotype.
PMID:
12382028 [PubMed - in process]
2:
Int J Radiat Oncol Biol Phys 2002
Nov 1;54(3):895
Fractionated
irradiation of H69 small-cell lung cancer cells causes
stable
radiation
and drug resistance with increased MRP1, MRP2, and
topoisomerase
IIalpha
expression.
Henness
S, Davey M, Harvie R, Davey R.
Bill
Walsh Cancer Research Laboratories, Medical Oncology Department,
Royal
PURPOSE:
After standard treatment with chemotherapy and radiotherapy,
small-cell
lung
cancer (SCLC) often develops resistance to both treatments. Our aims
were
to
establish if fractionated radiation treatment alone would induce
radiation
and
drug resistance in the H69 SCLC cell line, and to determine the
mechanisms
of
resistance.METHODS AND MATERIALS: H69 SCLC cells were treated
with
fractionated
X-rays to an accumulated dose of 37.5 Gy over 8 months to
produce
the
H69/R38 subline. Drug and radiation resistance was determined using the
MTT
(3,-4,5
dimethylthiazol-2,5 diphenyltetrazolium bromide) cell viability
assay.
Protein
expression was analyzed by Western blot.RESULTS: The H69/R38 subline
was
resistant
to radiation (2.0 +/- 0.2-fold, p < 0.0001), cisplatin (14 +/-
7-fold,
p
< 0.001), daunorubicin (6 +/- 3-fold, p < 0.05), and navelbine (1.7
+/-
0.15-fold,
p < 0.02). This was associated with increased expression of
the
multidrug
resistance-associated proteins, MRP1 and MRP2, and
topoisomerase
IIalpha
and decreased expression of glutathione-S-transferase pi (GSTpi)
and
bcl-2
and decreased cisplatin accumulation. Treatment with 4 Gy of
X-rays
produced
a 66% decrease in MRP2 in the H69 cells with no change in the
H69/R38
cells.
This treatment also caused a 5-fold increase in topoisomerase IIalpha
in
the
H69/R38 cells compared with a 1.5-fold increase in the H69
cells.CONCLUSIONS:
Fractionated radiation alone can lead to the development
of
stable
radiation and drug resistance and an altered response to radiation
in
SCLC
cells.
PMID:
12377343 [PubMed - in process]
3:
Blood 2002 Oct 15;100(8):2703-7
Negative
prognostic value of glutathione S-transferase (GSTM1 and
GSTT1)
deletions
in adult acute myeloid leukemia.
Voso
MT, D'Alo' F, Putzulu R, Mele L, Scardocci A, Chiusolo P, Latagliata
R,
Lo-Coco
F, Rutella S, Pagano L, Hohaus S, Leone G.
Istituto
di Ematologia, Universita' Cattolica S. Cuore and Dipartmento
di
Biotecnologie
Cellulari ed Ematologia, Universita' La Sapienza, Rome,
Glutathione
S-transferases (GSTs) are enzymes involved in the detoxification
of
several
environmental mutagens, carcinogens, and anticancer drugs.
GST
polymorphisms
resulting in decreased enzymatic activity have been
associated
with
several types of solid tumors. We determined the prognostic significance
of
the
deletion of 2 GST subfamilies genes, M1 and T1, in patients with
acute
myeloid
leukemia (AML). Using polymerase chain reactions, we analyzed the
GSTM1
and
GSTT1 genotype in 106 patients with AML (median age, 60.5 years;
range,
19-76
years). The relevance of GSTM1 and GSTT1 homozygous deletions was
studied
with
respect to patient characteristics, response to therapy, and
survival.
Homozygous
deletions resulting in null genotypes at the GSTM1 and GSTT1
loci
were
detected in 45 (42%) and 30 (28%) patients, respectively. The
double-null
genotype
was present in 19 patients (18%). GST deletions predicted poor
response
to
chemotherapy (P =.04) and shorter survival (P =.04). The presence of at
least
one
GST deletion proved to be an independent prognostic risk factor for
response
to
induction treatment and overall survival in a multivariate analysis
including
age
and karyotype (P =.02). GST genotyping was of particular prognostic value
in
the
cytogenetically defined intermediate-risk group (P =.003). In
conclusion,
individuals
with GSTM1 or GSTT1 deletions (or deletions of both) may have
an
enhanced
resistance to chemotherapy and a shorter survival. (Blood.
2002;100:2703-2707)
PMID:
12351375 [PubMed - in process]
4:
J Biol Chem 2002 Sep 23; [epub
ahead of print]
Neuronal
leucine-rich repeat protein-3 amplifies MAP kinase activation by
EGF
through
a carboxyl-terminal region containing endocytosis
motifs.
Fukamachi
K, Matsuoka Y, Ohno H, Hamaguchi T, Tsuda H.
Experimental
Pathology and Chemotherapy Division,
Research
Institute,
Neuronal
leucine-rich repeat protein-3 (NLRR-3) belongs to the LRR
superfamily.
We
have reported that rat NLRR-3 gene expression is regulated mainly through
the
Ras-mitogen-activated
protein kinase (MAPK) signaling pathway. NLRR-3 was found
to
enhance phosphorylation of MAPK when COS-7 cells were transfected with
NLRR-3
and
stimulated with a low concentration (0.01 ng/ml) of epidermal growth
factor
(EGF),
but the amplification of MAPK phosphorylation by NLRR-3 was no
longer
observed
when the carboxyl terminal 30 amino acid stretch that
contains
clathrin-mediated
endocytosis motifs was deleted. A green fluorescent
protein-tagged
NLRR-3 localized at the plasma membrane was efficiently
internalized
in COS-7 cells, but internalization of a carboxyl
terminal-deleted
version
(NLRR*C) was less efficient. The presence of clathrin-adaptor
protein
complexes
containing NLRR-3 in brain lysate was confirmed by
immunoprecipitation
and
glutathione S-transferase pull-down experiments, and affinity
column
chromatography
revealed that the carboxyl terminal region of NLRR-3
interacts
with
-adaptin. We propose that NLRR-3 potentiates Ras-MAPK signaling
by
facilitating
internalization of EGF in clathrin-coated vesicles.
PMID:
12297494 [PubMed - as supplied by publisher]
5:
Eur J Pharmacol 2002 Sep
13;451(2):203
Protection
by a radical scavenger edaravone against
cisplatin-induced
nephrotoxicity
in rats.
Sueishi
K, Mishima K, Makino K, Itoh Y, Tsuruya K, Hirakata H, Oishi
R.
Department
of Hospital Pharmacy, Faculty of Medicine,
Maidashi,
Higashi-ku, 812-8582,
Acute
renal failure is a dose-limiting factor of cisplatin chemotherapy.
Here,
we
show the protective effect of edaravone, a recently developed
radical
scavenger
for clinical use, against cisplatin-induced renal dysfunction in
rats.
A
marked increase in blood urea nitrogen and creatinine in serum,
and
histological
changes including vacuolation, necrosis and protein casts
were
observed
in proximal renal tubules at the fourth day after cisplatin
injection
(5-10
mg/kg). Repeated injection of edaravone (1-10 mg/kg, i.v. twice a day
for
3
days) reversed the cisplatin-induced elevation of blood urea nitrogen
and
creatinine,
and morphological changes in a dose-dependent manner. In
particular,
the
protective effect of edaravone was almost complete at 10 mg/kg.
Moreover,
the
compound was still fully effective, when it was administered only at
the
second
day after cisplatin injection. On the other hand, the glutathione
content
in
renal tissues lowered at the fourth day after cisplatin injection, which
was
reversed
by the late treatment with edaravone. These findings suggest that
the
clinically
available radical scavenger edaravone is potentially useful for
the
prevention
of cisplatin-induced renal toxicity.
PMID:
12231392 [PubMed - in process]
6:
J Biol Chem 2002 Sep 10; [epub
ahead of print]
Glutathione
influences c-Myc-induced apoptosis in M14 human melanoma
cells.
Biroccio
A, Benassi B, Filomeni G, Amodei S, Marchini S, Chiorino G, Rotilio
G,
Zupi
G, Ciriolo MR.
Department
of Experimental Chemotherapy Laboratory,
Institute,
The
present paper aims at dissecting the mechanisms by which the
down-regulation
of
c-Myc induces programmed cell death in melanoma cells. In stable
and
doxycycline-inducible
M14 melanoma cells, down-regulation of c-Myc induced
apoptosis
subsequent to a decrease in the intracellular reduced
glutathione
content
and a concomitant accumulation of its oxidized form. This
redox
alteration
was associated with a decrease of the enzyme activities of
g-glutamyl-cysteine
synthetase and NADPH-dependent GSSG-reductase, and to a
consequent
glutathione release in the extracellular medium. Cytochrome c
was
released
into the cytosol at very early stages of apoptosis induction,
long
before
detectable production of reactive oxygen species and activation
of
caspase-9
and -3. Macroarray analysis revealed that down-regulation of
c-Myc
produced
striking changes of gene expression in the section related
to
metabolism;
where the expression of g-glutamyl-cysteine synthetase and
GSSG-reductase
was found to be significantly reduced. Addition of
N-acetyl-L-cysteine
or glutathione ethyl-ester was able to inhibit the
apoptotic
process,
thus confirming the key role of glutathione in the programmed
cell
death
induced by c-Myc.
PMID:
12226097 [PubMed - as supplied by publisher]
7:
Cancer 2002 Aug 1;95(3):641-55
Evaluation
of the efficiency of chemotherapy in in vivo orthotopic models
of
human
glioma cells with and without 1p19q deletions and in C6 rat
orthotopic
allografts
serving for the evaluation of surgery combined with
chemotherapy.
Branle
F, Lefranc F, Camby I, Jeuken J, Geurts-Moespot A, Sprenger S, Sweep
F,
Kiss
R, Salmon I.
Department
of Oncology,
BACKGROUND:
Malignant gliomas of the central nervous system remain
associated
with
dismal prognoses because of their diffuse invasion of the brain
parenchyma.
Very
few experimental models that mimic clinical reality are available today
to
test
potentially new therapies. The authors set up experimental in vivo
glioma
models
of anaplastic astrocytomas of human and rat origins and
anaplastic
oligodendroglioma
of human origin. Standard hospital chemotherapies were
employed
to test the validity of these models. METHODS: Three glioma cells
lines
obtained
from the American Type Culture Collection (i.e., human Hs683 and
U373
cells
and rat C6 cells) were implanted into nude mouse brains (Hs683 and
U373
cells)
and rat brains (C6 cells). The astrocytic nature, as opposed to
the
oligodendrocytic
nature, of the Hs683 and U373 models was investigated by
using
quantitative
(computer-assisted microscopy) immunohistochemical
characterizations
of nestin, vimentin, glutathione-S-transferase alpha
(GSTalpha),
GSTmu, GSTpi, and p53 expression. Comparative genomic
hybridization
(CGH)
was employed to investigate 1p19q losses. Chronic administrations
of
carmustine
(BCNU), fotemustin, or temozolomide were assayed in the
xenografted
U373
and Hs683 models. Both BCNU-related chemotherapy and surgery were
assayed
in
the C6 model. RESULTS: The quantitative phenotypic analyses pointed to
the
oligodendroglial
nature of the Hs683 cell line and the astrocytic nature of
the
U373
cell line. The Hs683 cells exhibited 1p19q losses, whereas the U373
cells
did
not. BCNU, fotemustin, and temozolomide dramatically increased the time
of
survival
of the Hs683 oligodendroglioma-bearing mice, whereas temozolomide
only
induced
a weak but nevertheless statistically significant increase in the
U373
glioma-bearing
mice. In the C6 rat glioma model, surgery and BCNU
chemotherapy
were
more efficient than either treatment alone. CONCLUSIONS: The in vivo
models
of
gliomas of the central nervous system developed in the current work
best
mimicked
clinical reality. They can be used either to identify new
therapies
against
human gliomas or to optimize existing therapies. Copyright 2002
American
Cancer
Society.
PMID:
12209758 [PubMed - indexed for MEDLINE]
8:
Cancer Res 2002 Sep
1;62(17):4916-21
FK228
(depsipeptide) as a natural prodrug that inhibits class I
histone
deacetylases.
Furumai
R, Matsuyama A, Kobashi N, Lee KH, Nishiyama M, Nakajima H, Tanaka
A,
Komatsu
Y, Nishino N, Yoshida M, Horinouchi S.
Department
of Biotechnology, The
FK228
is a histone deacetylase (HDAC) inhibitor, the molecular mechanism
of
inhibition
of which has been unknown. Here we show that reduction of
an
intramolecular
disulfide bond of FK228 greatly enhanced its inhibitory
activity
and
that the disulfide bond was rapidly reduced in cells by cellular
reducing
activity
involving glutathione. Computer modeling suggests that one of
the
sulfhydryl
groups of the reduced form of FK228 (redFK) interacts with
the
active-site
zinc, preventing the access of the substrate. HDAC1 and HDAC2
were
more
strongly inhibited by redFK than HDAC4 and HDAC6. redFK was less
active
than
FK228 in inhibiting in vivo HDAC activity, due to rapid inactivation
in
medium
and serum. Thus, FK228 serves as a stable prodrug to inhibit class
I
enzymes
and is activated by reduction after uptake into the cells.
The
glutathione-mediated
activation also implicates its clinical usefulness for
counteracting
glutathione-mediated drug resistance in chemotherapy.
PMID:
12208741 [PubMed - indexed for MEDLINE]
9:
Bone Marrow Transplant 2002
Aug;30(3):135-40
Synergistic
cytotoxicity of buthionine sulfoximine (BSO) and intensive
melphalan
(L-PAM)
for neuroblastoma cell lines established at relapse after
myeloablative
therapy.
Division
of Hematology-Oncology,
Patients
with high-risk neuroblastoma (NB) initially respond to
aggressive,
alkylator-based
therapy only to die from recurrent disease that is refractory
to
chemotherapy,
including alkylating agents. We examined the ability of
buthionine
sulfoximine
(BSO)-mediated glutathione (GSH) depletion to modulate
melphalan
(L-PAM)
resistance in five NB cell lines established after progressive
disease
following
myeloablative therapy (high-dose melphalan, carboplatin, etoposide
and
total
body irradiation) supported by autologous hematopoietic stem
cell
transplant
(AHSCT), and in 15 NB cell lines established at diagnosis or
after
non-myeloablative
therapy (pre-AHSCT). Four of five post-AHSCT NB cell lines
and
10
of 15 pre-AHSCT NB cell lines were sensitive to single agent BSO (LC(90)
<300
microM
BSO), while two of five post-AHSCT lines and one of 15 pre-AHSCT
lines
showed
high-level resistance to L-PAM (LC(90)>30 microM). Fixed ratio
analysis
demonstrated
BSO/L-PAM synergy (combination index <1) for all five
post-AHSCT
and
for all 15 pre-AHSCT cell lines tested. Multi-log cytotoxicity
(often
exceeding
four logs of cell kill) was observed in post-AHSCT
L-PAM-resistant
cell
lines (including p53 non-functional lines) only when clinically
achievable
concentrations
of BSO were combined with myeloablative concentrations of
L-PAM.
We
conclude that most neuroblastoma cell lines, including post-AHSCT NB
cell
lines
that are highly resistant to myeloablative levels of L-PAM and lack
p53
function,
are sensitive to clinically achievable concentrations of L-PAM
and
BSO.
However, some L-PAM-resistant NB cell lines (especially those lacking
p53
function)
require dose escalation of L-PAM to myeloablative concentrations
in
order
to demonstrate significant synergistic cytotoxicity. Thus,
optimal
clinical
application of BSO/L-PAM may require AHSCT.
PMID:
12189530 [PubMed - in process]
10:
Br J Cancer 2002 Jul 29;87(3):251-7
Protein
expression profiles indicative for drug resistance of non-small
cell
lung
cancer.
Volm
M, Koomagi R, Mattern J, Efferth T.
German
Cancer Research Centre,
Data
obtained from multiple sources indicate that no single mechanism
can
explain
the resistance to chemotherapy exhibited by non-small cell
lung
carcinomas.
The multi-factorial nature of drug resistance implies that
the
analysis
of comprising expression profiles may predict drug resistance
with
higher
accuracy than single gene or protein expression studies. Forty
cellular
parameters
(drug resistance proteins, proliferative, apoptotic, and
angiogenic
factors,
products of proto-oncogenes, and suppressor genes) were
evaluated
mainly
by immunohistochemistry in specimens of primary non-small cell
lung
carcinoma
of 94 patients and compared with the response of the tumours
to
doxorubicin
in vitro. The protein expression profile of non-small cell
lung
carcinoma
was determined by hierarchical cluster analysis and clustered
image
mapping.
The cluster analysis revealed three different resistance profiles.
The
frequency
of each profile was different (77, 14 and 9%, respectively). In
the
most
frequent drug resistance profile, the resistance proteins
P-glycoprotein/MDR1
(MDR1, ABCB1), thymidylate-synthetase,
glutathione-S-transferase-pi,
metallothionein,
O6-methylguanine-DNA-methyltransferase
and major vault protein/lung
resistance-related
protein were up-regulated. Microvessel density, the
angiogenic
factor vascular endothelial growth factor and its receptor FLT1,
and
ECGF1
as well were down-regulated. In addition, the proliferative
factors
proliferating
cell nuclear antigen and cyclin A were reduced compared to
the
sensitive
non-small cell lung carcinoma. In this resistance profile, FOS
was
up-regulated
and NM23 down-regulated. In the second profile, only three
resistance
proteins were increased (glutathione-S-transferase-pi,
O6-methylguanine-DNA-methyltransferase,
major vault protein/lung
resistance-related
protein). The angiogenic factors were reduced. In the
third
profile,
only five of the resistance factors were increased (MDR1,
thymidylate-synthetase,
glutathione-S-transferase-pi,
O6-methylguanine-DNA-methyltransferase,
major vault protein/lung
resistance-related
protein). Copyright 2002 Cancer Research
PMID:
12177790 [PubMed - indexed for MEDLINE]
11:
J Clin Oncol 2002 Aug
15;20(16):3478-83
Neuroprotective
effect of reduced glutathione on oxaliplatin-based
chemotherapy
in
advanced colorectal cancer: a randomized, double-blind,
placebo-controlled
trial.
Cascinu
S, Catalano V, Cordella L, Labianca R, Giordani P, Baldelli AM,
Beretta
GD,
Ubiali E, Catalano G.
Department
of Medical Oncology, Azienda Ospedaliera-Universitaria di
Gramsci
14, 43100
PURPOSE:
We performed a randomized, double-blind, placebo-controlled trial
to
assess
the efficacy of glutathione (GSH) in the prevention of
oxaliplatin-induced
neurotoxicity. PATIENTS AND METHODS: Fifty-two patients
treated
with a bimonthly oxaliplatin-based regimen were randomized to
receive
GSH
(1,500 mg/m(2) over a 15-minute infusion period before oxaliplatin)
or
normal
saline solution. Clinical neurologic evaluation and
electrophysiologic
investigations
were performed at baseline and after four (oxaliplatin dose,
400
mg/m(2)),
eight (oxaliplatin dose, 800 mg/m(2)), and 12 (oxaliplatin dose,
1,200
mg/m(2))
cycles of treatment. RESULTS: At the fourth cycle, seven
patients
showed
clinically evident neuropathy in the GSH arm, whereas 11 patients in
the
placebo
arm did. After the eighth cycle, nine of 21 assessable patients in
the
GSH
arm suffered from neurotoxicity compared with 15 of 19 in the placebo
arm.
With
regard to grade 2 to 4 National Cancer Institute common toxicity
criteria,
11
patients experienced neuropathy in the placebo arm compared with only
two
patients
in the GSH arm (P =.003). After 12 cycles, grade
was
observed in three patients in the GSH arm and in eight patients in
the
placebo
arm (P =.004). The neurophysiologic investigations (sural sensory
nerve
conduction)
showed a statistically significant reduction of the values in
the
placebo
arm but not in the GSH arm. The response rate was 26.9% in the GSH
arm
and
23.1% in the placebo arm, showing no reduction in activity of
oxaliplatin.
CONCLUSION:
This study provides evidence that GSH is a promising drug for
the
prevention
of oxaliplatin-induced neuropathy, and that it does not reduce
the
clinical
activity of oxaliplatin.
Publication
Types:
Clinical
Trial
Randomized
Controlled Trial
PMID:
12177109 [PubMed - indexed for MEDLINE]
12:
Curr Top Med Chem 2002
Jul;2(11):1239-59
Enzymes
of the trypanothione metabolism as targets for antitrypanosomal
drug
development.
Schmidt
A, Krauth-Siegel RL.
Biochemie-Zentrum
328,
Trypanosomatids
are the causative agents of African sleeping sickness,
Chagas'
disease
and the three forms of leihmaniasis. New drugs against these
parasitic
protozoa
are urgently needed since the currently available chemotherapy is
not
at
all satisfying. One promising approach towards the development of new
drugs
is
based on the design of specific enzyme inhibitors. Trypanosomes
and
leishmania
possess a unique thiol metabolism in which the ubiquitous
glutathione/glutathione
reductase system is replaced by trypanothione and
trypanothione
reductase. The dithiol trypanothione is the key molecule for
the
synthesis
of DNA precursors, the homeostasis of ascorbate, the detoxification
of
hydroperoxides,
and the sequestration/export of thiol conjugates. Synthesis
and
reduction
of trypanothione are essential for the maintenance of a
reducing
intracellular
milieu which renders the respective enzymes attractive drug
target
molecules.
Here we present the current state of knowledge of the
thiol
metabolism
of trypanosomatids, comprising the
trypanothione/tryparedoxin,
thioredoxin,
and ovothiol systems of the parasites. The most effective
inhibitors
of the enzymes known to date, their mode of action, and
the
(dis)advantages
of different types of inhibitors as potential drug
candidates
will
be discussed.
PMID:
12171583 [PubMed - in process]
13:
Clin Exp Med 2002 Jul;2(2):99-103
Modulation
of intracellular glutathione concentration alters
dehydropyrimidine
dehydrogenase
activity in peripheral blood mononuclear cells.
Iwata
S, Takabayashi A, Yamaoka Y.
Department
of Gastroenterological Surgery and Oncology,
Tazuke
Kohukai Medical Research Institute,
iwata@hosp.city.kyoto.jp
Dehydropyrimidine
dehydrogenase (DPD) is the initial key enzyme in the
regulation
of 5-fluorouracil catabolism and thus controls availability
of
5-fluorouracil
for anabolism. Modulation of DPD activity may increase the
antitumor
effect and avoid toxic side effects in
5-fluo-rouracil-based
chemotherapy.
We measured DPD activity in peripheral blood mononuclear
cells
from
cancer patients and simultaneously monitored intracellular
glutathione
(GSH) and plasma GSH l