Download Mushroom Lectin Inhibits Invasion of a Human Colon Cancer Cell

I2P
Medical Research Society
marked inhibition of c-myc expression by the anti-gal galNAc
unexpected and suggests that c-myc activation is not essential for
proliferation in this cell line.
control
ET-1,
lO0nw
PEP.
0
0.5
1
6
24
.07+.03
.18+.02
.06+.03
.17+.09
.19+.10
.22+.07
.66+.18*
.87+.20'
.97+.18'
.13+.04
.59+.12*
.94+.09t
.89+.21*
loom
NM
MUSHROOM LECTIN INHIBITS INVASION OF A
HUMAN COLON CANCER CELL LINE INTO COLLAGEN
Conclusions Ventricular myocytes produce ET-1 rnRNA but
not ET-2 or ET-3 rnRNAs. PtP and ET-1 increase ET-1 mRNA
levels in myocytes. PEP-induced hypertrophy may be
mediated at least in part by ET-1 generation from
myocytes .
GELS
RC EVANS. JM RHODES and AR KINSELLA'
N42
Departments of Medicine and Surgery', University of Liverpool,
PO Box 147, Liverpool L69 3BX
The edible mushroom (Aguricw bispom) contains a lectin
which binds the oncofclal antigen Gal513GalNAc and inhibits
proliferation of cancer cells ( Cancer Res 1993; 53: 4627-32 ). Its
potential as an anticancer therapy has now been assessed further
by studying its effect on the migration of an invasive, E-Cadherin
negative, clone. (2010/1) of the HCT 116 human colon cancer cell
line.
Method: The invasive cells were seeded at 10s celldml onto
a m s t i t u t e d collagen matrix. Mushroom lectin was added to the
media at 30Fglml. Inverted micmmpy was used to count the
invading cells daily over 4 days and this count was expressed as
a percentage of the cells remaining on the cell surface. At each
time period blinded counts were made of 10 fielddwell. in
triplicate and compand lo control wells.
Results: Invasion was inhibited in the mushroom lectin
containing wells, by 28.0% @=0.008) on day 3 and by 52.8%
@<O.oOol) by day 4.
Mushroom lectin inhibits tumour cell invasion in addition to
inhibition of proliferation and deserves further evaluation as a
possible anticancer agent.
N41 CULTURED VENTRICMYOCYTES EXPRESS ENDOTEELIN-1
(ET-1) MRNA BUT NOT ET-2 MRNA OR ET-3 MRNA IN RESPONSE
TO TEE EYPERTROPEIC AGONISTS PEENYLEPERINE AND ET-1
S KADWVRA, J FIRTE.,
S J FULLER, P A POOLE-WILSON,
P E SUGDEN (INTRODUCED BY P A POOLE-WILSON)
National Eeart and Lung Institute, London
*Institute of Molecular Medicine, Oxford, UR
and
k e c k a r o u In addition to its potent vasoactive
properties, endothelin-1 (ET-1) has powerful effects on
cell growth end may be involved in the process of
myocardial hypertrophy. The roles of ET-2 and ET-3 in
this process are unknown. We have investigated the
effects of exogenous ET-1 and the a-adrenergic agonist
phenylephrine (PEP) on levels of ET-1, ET-2 and ET-3
mRNAs in myocytes.
Kethode Neonatal rat ventricular myocytes were
8timulated with ET-1 or PtP. Quantitative ribonuclease
protection assay and laser denmitometry rere used to
assay levels Of the mRNAs for ET-1, ET-2 and tT-3, and
constitutive WDE.
Result. tT-1 mRNA was present in u n s t h l a t e d myocytes.
ET-2 and ET-3 m R N h e r e not detectable in contol or
stimulated myocytes.
ET-1 and PEP caused a dosedependent increase in --1 ~ R N A relative to GAPDE ~ R N A
within 60 minutes. ET-1 mRNA levels remained elevated
to 24 hours:
TISSUE
ENDOTOXAEMIA
EXPRESSION
OF
ENDOTEELIN-1
MRNA
IN
S KADWDRA, NP CURZEN, J FIRTE., PE SUGDEN, PA POOLEWILSON AND TW EVANS (INTRODUnD BY PA POOLE-WILSON)
National Eeart and Lung Institute, London
*Institute of Uolecular Medicine, Oxford, UK
and
Backaround Circulating endothelin-1 concentrations
([ET-11) are elevated in animal models of sepsis and in
critically ill patients, in whom they correlate with
outcome (Ann S u r g 1991;213:261-4). The main actions of
ET-1 appear to be as a local autocrine and paracrine
factor, rather than as a circulating hormone.
Plasma
[ET-I] may not be a true reflection of local tissue
concentrations. We measured tissue [ET-1 mRNA] as an
indication of local production in endotoxaemia.
27 adult male wistar rats were divided into 3
groups: [l] Control ( C ) - intraperitoneal (ip) injection
of saline, ( 2 1 Endotoxin (E)- ?Omg/kg ip endotoxin, [.3]
30 mins
Dexamethnsone (D)- 3mglkg ip dexmethasone
prior to endotoxin. Rats were killed by cervical
dislocation at 1, 4 and 6h. Tissues were rapidly
removed, frozen and ground under liquid NI. Total RNA
w a 8 extracted by the guanidinium thiocyanate method. A
quantitative ribonucleaae protection assay was
performed on 25pg of RNA from each tissue using a
specific probe protecting 154 bases of ET-1 rnRNA, and a
probe protecting 316 bases of constitutive GAPDE mRNA.
Protected bands were quantified by laser densitamtry.
pesule There was a tissue-specific increase in tissue
ET-1 mRNA expression in enSotoxaesnia:
Tissue ET-1 mRNA/GAPDE mRNA ratio, 6 hours
tiasue
control
endotoxin
dexamethawne
0.204f0.022
0.356f0.014t
heart
0.290i0.005*1.506f0.383
3.030i0.204.
2.596f0.226
lung
0.813f0.012
1.055f0.093
kidney
0.893f0.027
0.209f0.024
skeletal mus 0.261fO.034
0.301f0.029
(means f sen, n-3. Relative to C:
p<O.O5, tpi0.01,
and for D relative to E: 3 p-0.01).
Conclusions Endotoxin causes differential increases in
tissue [ET-1 mRNA] in vivo. This is most pronounced in
lung and heart and may account for some of the adverse
effects o f endotoxaemia
upon
these
organs.
Dexamethasone causes some suppression of the rise in
[ET-lmRNA] but at 6h this is only significant in heart.
NO
A NINE-MINUTE WALK TEST ON A PATIENT-POWERED
TREADMILL DURING TREATMENT OF SEVERE EEART FAILURE;
EXERCISE CAPACITY AND RELATION TO SYMPTOMS AND WEIGET
0 PATEL,
J BAYLISS',
G C SUTTON*',
BY P A POOLG-WILSON)
S KADDOURA,
J PARMIESEWAR,
J SPARROW,
P A POOLE-WILSON (INTRODUCED
Royal Brompton Eospital, London; 'Eemel
Eospital and **Eillingdon Eospital, UR
Eempstead
Changes in time-limited exercise capacity
on a
patient-powered (Tunturi) treadmill were assessed
during in-patient treatment of 10 patients with
decunpensated chronic heart failure (NYEA classes 111
and IV). Patients carried out a 9-minute walk on the