Journal Medicine: The New England

Vol ume 339 Number 12
785

The New England

Journal

of

Medicine

Copyr i ght, 1998, by t he Massachus et t s Medi cal Soci et y

VOLUME 339

S

EPTEMBER

17, 1998

NUMBER 12

CLINICAL AND BIOLOGIC ACTIVITY OF AN ESTROGENIC HERBAL
COMBINATION (PC-SPES) IN PROSTATE CANCER

R

OBERT

S. D

I

P

AOLA

, M.D., H

UAYAN

Z

HANG

, M.D., G

EORGE

H. L

AMBERT

, M.D., R

OBERT

M

EEKER

, B.S.,
E

DWARD

L

ICITRA

, P

H

.D., M

OHAMED

M. R

AFI

, P

H

.D., B

AO

T

ING

Z

HU

, P

H

.D., H

EIDI

S

PAULDING

, R.N.,
S

USAN

G

OODIN

, P

HARM

.D., M

ICHEL

B. T

OLEDANO

, M.D., W

ILLIAM

N. H

AIT

, M.D., P

H

.D.,

AND

M

ICHAEL

A. G

ALLO

, P

H

.D.

A

BSTRACT

Background

Herbal mixtures are popular alterna-
tives to demonstrated therapies. PC-SPES, a commer-
cially available combination of eight herbs, is used
as a nonestrogenic treatment for cancer of the pros-
tate. Since other herbal medicines have estrogenic
effects in vitro, we tested the estrogenic activity of
PC-SPES in yeast and mice and in men with prostate
cancer.

Methods

We measured the estrogenic activity of
PC-SPES with transcriptional-activation assays in
yeast and a biologic assay in mice. We assessed the
clinical activity of PC-SPES in eight patients with hor-
mone-sensitive prostate cancer by measuring serum
prostate-specific antigen and testosterone concen-
trations during and after treatment.

Results

In complementary yeast assays, a 1:200
dilution of an ethanol extract of PC-SPES had estro-
genic activity similar to that of 1 nM estradiol, and in
ovariectomized CD-1 mice, the herbal mixture in-
creased uterine weights substantially. In six of six men
with prostate cancer, PC-SPES decreased serum tes-
tosterone concentrations (P<0.05), and in eight of
eight patients it decreased serum concentrations of
prostate-specific antigen. All eight patients had breast
tenderness and loss of libido, and one had venous
thrombosis. High-performance liquid chromatogra-
phy, gas chromatography, and mass spectrometry
showed that PC-SPES contains estrogenic organic
compounds that are distinct from diethylstilbestrol,
estrone, and estradiol.

Conclusions

PC-SPES has potent estrogenic ac-
tivity. The use of this unregulated mixture of herbs
may confound the results of standard or experimen-
tal therapies and may produce clinically significant
adverse effects. (N Engl J Med 1998;339:785-91.)

1998, Massachusetts Medical Society.

From the Departments of Medicine (R.S.D., E.L., M.M.R., B.T.Z.,
H.S., S.G., W.N.H., M.A.G.), Pediatrics (H.Z., G.H.L.), and Pharmacology
(W.N.H.), University of Medicine and Dentistry of New JerseyRobert
Wood Johnson Medical School, New Brunswick; the Cancer Institute of
New Jersey, New Brunswick (R.S.D., G.H.L., R.M., E.L., M.M.R.,
B.T.Z., H.S., S.G., W.N.H., M.A.G.); and the Environmental and Occupa-
tional Health Sciences Institute, Piscataway, N.J. (H.Z., G.H.L., R.M.,
B.T.Z., M.B.T., M.A.G.). Address reprint requests to Dr. DiPaola at the
Cancer Institute of New Jersey, University of Medicine and Dentistry of
New JerseyRobert Wood Johnson Medical School, 195 Little Albany St.,
New Brunswick, NJ 08901.

ERBAL therapies are unconventional
treatments in wide use for many diseases.
They are sold as nutritional supplements
for numerous illnesses, including the com-
mon cold (echinacea),

1

benign prostatic hypertrophy
(saw palmetto),

2

and depression (Saint Johnswort).

3

Among patients with cancer, the use of unconven-
tional medicines, including herbal therapies, has been
reported to be as low as 5 percent and as high as 60
percent.

4,5

PC-SPES is an herbal combination used
by patients with prostate cancer that consists of eight
herbs: chrysanthemum, isatis, licorice,

Ganoderma
lucidum, Panax pseudo-ginseng, Rabdosia rubescens,

saw palmetto, and scutellaria (skullcap).

6-8

Herbal therapies can have important biologic activ-
ities. For example, saw palmetto inhibits 5

a

-reduc-
tase, an enzyme involved in testosterone metabolism,

9

and Saint Johnswort, like pharmacologic antidepres-
sants, blocks monoamine oxidase activity.

10

Although
PC-SPES is promoted as a nonestrogenic food sup-
plement, some of its constituents have estrogenic
activity.

6-8

Licorice competes with estradiol in an es-
trogen-receptorbinding assay,

11

and ginseng induces
estrogen-regulated expression of pS2, a small protein
found in breast cancer, in cultured MCF-7 breast-
cancer cells.

12

The clinical implications of such activ-
ities are unknown.
H
The New England Journal of Medicine
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Copyright 1998 Massachusetts Medical Society. All rights reserved.

786

September 17, 1998

The New Engl and Jour nal of Medi ci ne

Figure 1.

Effect of PC-SPES and Estradiol on

b

-Galactosidase
Activity of Y253 Yeast with a

lacZ

Construct Downstream of a
Human Estrogen-Response Element.
Both PC-SPES (Panel A) and estradiol (Panel B) activated the

lacZ

gene, as measured by

b

-galactosidase activity. PC-SPES
had a dilution-dependent effect that was similar to the effect of
estradiol. Each point represents the mean (SE) of four separate
experiments with estradiol and batches A, B, and C of PC-SPES.
0
600
0.1
100
200
300
400
500
0.01 0.001 0.0001 0.00001 0
PC-SPES (1/dilution)
b
-
G
a
l
a
c
t
o
s
i
d
a
s
e

A
c
t
i
v
i
t
y

(
U
)
0
800
100 10 1
200
400
600
0.1 0.01 0.001 0.0001 0
Estradiol (nM)
b
-
G
a
l
a
c
t
o
s
i
d
a
s
e

A
c
t
i
v
i
t
y

(
U
)

We recently observed that PC-SPES had clinical ac-
tivity in a man with prostate cancer that had recurred
after radical prostatectomy. His prostate-specific an-
tigen concentration was 34 ng per milliliter when he
started taking PC-SPES, without concurrent thera-
pies. After one month, he reported breast tenderness
and loss of libido, and his prostate-specific antigen
had decreased to 0.4 ng per milliliter. These findings
prompted a study to determine the mechanism of
these effects. Our results indicate that PC-SPES has
estrogenic activity, reduces concentrations of serum
testosterone, exerts activity against prostate cancer,
and causes untoward side effects.

METHODS

Materials

PC-SPES was purchased from Botaniclab (Brea, Calif.) in the
form of 320-mg capsules for laboratory and clinical studies. To
determine whether the composition of PC-SPES varied from lot
to lot, we made four separate purchases of PC-SPES and analyzed
each batch. Ginseng (Action Labs, Long Island, N.Y.) and saw
palmetto (Fingerprint Botanicals, General Nutrition, Pittsburgh)
were purchased from a health-food vendor. Stock solutions of
PC-SPES and herbal extracts were prepared by exposing them to
ethanol (dilution, 1:10 wt/vol) for 24 hours. Estrone, 17

b

-estradi-
ol, and diethylstilbestrol were purchased from Sigma Chemical
(St. Louis). Ovariectomized CD-1 mice (weight, 18 to 20 g) were
purchased from Charles River Laboratories (Wilmington, Mass.)
and housed under standard conditions in the Robert Wood
Johnson Medical School vivarium.

Estrogen-Receptor Activity and Estrogen-Dependent
Growth

The estrogenic activity of PC-SPES was assessed with a

b

-galac-
tosidase assay with the yeast strain Y253

(ura352,his3

200,

leu2

1,

lys2801,ade2101,yap1::leu2).

In Y253, two plasmids
were introduced. One is the plasmid YEp90HEGO, which express-
es the human estrogen receptor

a

. The other plasmid (PYER

1

)
carries the

lacZ

gene, which encodes the enzyme

b

-galactosidase.
In this plasmid, the upstream activating sequence of the gene was
replaced by two estrogen-response elements. In this system,

b

-galactosidase activity is proportional to the degree of activation
of estrogen-response elements.

13

A complementary growth-based assay used the

Saccharomyces
cerevisiae

strain PL3

(ura3

1,

his3

200,

leu2

1,

trp1::3ERE
URA3

), which was a gift of Dr. R. Losson.

13

PL3 carries a

URA3

gene that is under the control of the estrogen-response element.
Transcription of

URA3,

which is required for the growth of the
cells in medium lacking uracil, is dependent on the activation of
the human estrogen receptor by a ligand. Cells were seeded in
96-well plates in 190 l of medium lacking uracil. Serial dilutions
of 10 l of 17

b

-estradiol or PC-SPES extract were added to the
cultures, and growth was observed for four days.

Studies in Animals

Ovariectomized CD-1 mice received 1 ml of either a suspen-
sion of PC-SPES (five mice) or vehicle alone (five) orally on days
0, 1, 2, and 3. The suspension contained five capsules of PC-
SPES ground into powder and suspended in 20 ml of an aqueous
solution containing 5 percent gum arabic. Three additional ani-
mals received no treatment throughout the course of the exper-
iment. On day 4, the animals were weighed and then killed by
cervical dislocation. The uteri were removed and weighed. All
studies in animals were approved by the institutional animal care
and use committee of the Robert Wood Johnson Medical
School.

High-Performance Liquid Chromatography, Gas
Chromatography, and Mass Spectrometry

PC-SPES capsules were treated with 5 ml of 100 percent eth-
anol, the insoluble herbal residues were precipitated by centrifu-
gation at 3500 rpm for 20 minutes, and the ethanol extract was
dried under a stream of nitrogen. The dried residues were dis-
solved in 1 ml of ethanol and filtered through 0.45-m cellulose

B
A

CLI NI CAL AND BI OLOGI C ACTI VI TY OF AN ESTROGENI C HERBAL COMBI NATI ON ( PC- SPES) I N PROSTATE CANCER

Vol ume 339 Number 12

787

Figure 2.

Effect of PC-SPES on the Growth of the PL3 Strain of

S. cerevisiae.

In the PL3 strain the

URA3

gene is under the control of a human
estrogen receptor, and this gene is required for the growth of
cells in uracil-deficient medium. Growth is observed in the pres-
ence of ethanol-extracted PC-SPES (1:200) from four separate
batches and 1 nM estradiol three days after 2000 cells (lane 1),
1000 cells (lane 2), and 500 cells (lane 3) had been plated. The
negative control consists of ethanol in uracil-deficient agar. The
positive control consists of ethanol in agar with uracil. The re-
sults are representative of four separate experiments.
Negative control
Positive control
PC-SPES, batch A
Estradiol
1 2 3
PC-SPES, batch B
PC-SPES, batch C
PC-SPES, batch D

acetate membranes, and 40-l aliquots were analyzed by high-
performance liquid chromatography as previously described.

14,15

Elutants were collected from the column, dried under a stream
of nitrogen, and analyzed by gas chromatography and mass spec-
trometry. The dried elutants were redissolved in 30 l of ethyl
acetate and exposed to 50 l of bis(trimethylsilyl)trifluoroaceta-
mide at 68C for 45 minutes. Gas chromatography and mass
spectrometry were performed on a Varian Star gas chromato-
graph (model 3400, Varian, Walnut Creek, Calif.) coupled with a
Varian Saturn ion-trap mass spectrometer (model 2000, Varian).
A 30 m by 0.32 mm by 0.25 m DB-5 capillary column (J and
W Scientific, Folsom, Calif.) was used for the separation. The
3-l sample was injected into the gas chromatograph at a temper-
ature of 280C. The column temperature was programmed to in-
crease from 35C to 300C over a period of 55 minutes. The
mass spectrometer was set to scan particles with a mass-to-charge
ratio ranging from 17 to 600 every 0.6 second.

Clinical Studies

We studied eight patients: seven were enrolled in an observa-
tional study approved by the institutional review board at Robert
Wood Johnson Medical School, and one met eligibility require-
ments before the study began (Patient 1). All eight patients had
histologically proved prostate cancer, without progression during
previous androgen-ablation therapy. The patients were required
to have taken PC-SPES for at least one month and to have taken
a minimum of four 320-mg capsules of PC-SPES daily for two
weeks. During the study they could not receive any standard
forms of androgen-ablation therapy. The effects of PC-SPES were
assessed with a standardized questionnaire. Serum prostate-spe-
cific antigen and testosterone concentrations were measured
while the patients were taking PC-SPES and two to six weeks af-
ter they had stopped taking the preparation. Pretreatment pros-
tate-specific antigen concentrations were obtained from the pa-
tients records.

Statistical Analysis

Data are presented as means SE. The uterine weights in the an-
imals were compared with use of the Wilcoxon rank-sum test. The
patients prostate-specific antigen and testosterone values were com-
pared with use of the Wilcoxon signed-rank test. All P values were
two-sided.

RESULTS

Estrogenic Activity in Yeast

We used two yeast strains to measure the estrogenic
activity of PC-SPES. First, Y253 yeast harboring the

lacZ

gene under the control of the human estrogen
receptor was treated with PC-SPES or estradiol. The
PC-SPES extract caused a dose-dependent increase
in

b

-galactosidase activity (Fig. 1A). A 1:200 dilu-
tion of the extract had estrogenic activity equivalent
to that of 1 nM estradiol (Fig. 1).
We next analyzed the effects of estradiol and PC-
SPES on the growth of the estrogen-dependent

S. cerevisiae

strain PL3. An ethanol extract from
four separate batches of PC-SPES (1:200 dilution) or
1 nM estradiol supported the growth of PL3 in uracil-
deficient medium (Fig. 2). Table 1 shows that all four
batches of highly dilute PC-SPES extract (1:20,000)
supported the growth of PL3. Estradiol supported
growth of the yeast at a minimal concentration of
110

5

nM. Growth was not supported by herbal
extracts of saw palmetto or ginseng.

Estrogenic Activity in Animals

To determine whether PC-SPES has estrogenic ac-
tivity in animals, we treated ovariectomized CD-1
mice with 1 ml of a suspension of PC-SPES or vehicle
daily for four days. As shown in Table 2, animals given
PC-SPES had a mean uterine weight of 76.912.1
mg, as compared with a weight of 18.53.2 mg in
mice given vehicle (P=0.008) and 19.52.8 mg in
untreated animals.

Effect of PC-SPES on Serum Testosterone and Prostate-
Specific Antigen in Patients with Prostate Cancer

We evaluated the activity of PC-SPES in eight pa-
tients with biopsy-proved prostate cancer before and
during treatment with PC-SPES and in six patients

788

September 17, 1998


two to six weeks after treatment was stopped (Table
3). Serum testosterone concentrations decreased dur-
ing the use of PC-SPES and increased within three
weeks after PC-SPES was discontinued (P<0.05)
(Table 3). The mean testosterone concentration was
237.081 ng per deciliter (822281 nmol per liter;
range, 25 to 494 ng per deciliter [87 to 1713 nmol
per liter]) during treatment and 879144 ng per
deciliter (3047499 nmol per liter; range, 301 to
1324 ng per deciliter [1044 to 4590 nmol per liter])
after treatment was discontinued. The serum testos-
terone concentration was below the normal reference
range in four of the eight patients, and in two patients
(Patients 1 and 3 in Table 3) it was below 30 ng per
deciliter (104 nmol per liter), a concentration similar
to that in patients receiving a standard regimen of an-
drogen-ablation therapy.

16

Serum testosterone con-
centrations were within the normal reference range in
four patients during treatment with PC-SPES (Pa-
tients 4, 5, 6, and 8 in Table 3) and increased within
three weeks if the treatment was discontinued.
The serum prostate-specific antigen concentration
was assessed before and during the use of PC-SPES.
It decreased in all eight patients after the initiation
of treatment with PC-SPES (Table 3), whether the
initial concentration was high (as in Patients 1 and
3) or low (as in Patient 8). The concentration began
to increase within three weeks after PC-SPES was
discontinued (Table 3).
Other clinical effects observed during PC-SPES
use were similar to those observed with pharmaco-
logic doses of estrogen.

16-18

All eight patients had
breast tenderness and loss of libido. One patient had
a superficial venous thrombosis after taking nine
capsules of PC-SPES per day for more than one
month. No other serious adverse effects were noted.

Composition of PC-SPES

We used high-performance liquid chromatogra-
phy to analyze the composition of PC-SPES. Repre-
sentative chromatograms for estrone, estradiol, and
diethylstilbestrol are shown in Figure 3A. High-per-
formance liquid chromatography of PC-SPES dem-
onstrated four peaks over a period of 53 to 64 min-
utes (Fig. 3B). Elutants corresponding to each of
these four peaks had estrogenic activity in the yeast-
growth assay (data not shown).
The peaks representing estrone and estradiol (Fig.
3A) differed from the major peaks found in PC-
SPES (Fig. 3B). Since peak X of PC-SPES resembled
that of diethylstilbestrol, we analyzed this peak using
gas chromatography and mass spectrometry. Peak X
elutants contained a mixture of organic chemicals
(Fig. 3D). Among them, only one peak (labeled
peak Y in Fig. 3D) resembled diethylstilbestrol (Fig.
3C). In-line mass spectrometric analysis of this peak
(inset in Fig. 3D) showed that it differed from that
of diethylstilbestrol (inset in Fig. 3C).

DISCUSSION

We found that PC-SPES, an unregulated herbal
dietary supplement, has potent estrogenic activity in
yeast, mice, and humans. In patients with prostate
cancer, it causes clinically significant reductions in se-
rum testosterone concentrations, decreases in pros-
tate-specific antigen concentrations, and side effects
similar to those of pharmacologic doses of estrogen.
In all eight patients we studied, PC-SPES reduced
serum testosterone concentrations to levels shown in
the randomized trial by the Leuprolide Study Group
to have antitumor activity.

16

PC-SPES also reduced

T

ABLE 2. EFFECT OF PC-SPES IN MICE.
TREATMENT AND
MOUSE NO.
FINAL
BODY
WEIGHT
UTERINE
WEIGHT
UTERINE
WEIGHT/
BODY
WEIGHT
g mg
PC-SPES
1
2
3
4
5
Mean SE
18.9
19.8
22.0
21.8
23.7
21.20.9
68.9
37.4
79.8
86.6
111.8
76.912.1
3.65
1.89
3.63
3.97
4.72
3.60.5
Vehicle
6
7
8
9
10
Mean SE
17.3
22.4
17.5
23.2
18.9
19.91.2
29.2
11.6
15.1
14.5
22.2
18.53.2
1.69
0.52
0.86
0.62
1.17
1.00.2
No treatment
11
12
13
Mean SE
22.5
22.1
23.3
22.60.4
20.6
23.8
14.2
19.52.8
0.92
1.08
0.61
0.90.14
*NT denotes not tested.
TABLE 1. EFFECT OF HERBAL EXTRACTS AND ESTRADIOL
ON THE GROWTH OF ESTROGEN-DEPENDENT YEAST.*
ESTRADIOL
CONCENTRATION
(nM) GROWTH
HERBAL-
EXTRACT
DILUTION GROWTH
PC-SPES BATCH GINSENG
SAW
PALMETTO
A B C D
1 + 1:210
1
+ + + +
110
1
+ 1:210
2
+ + + +
110
2
+ 1:210
3
+ + + +
110
3
+ 1:210
4
+ + + +
110
4
+ 1:210
5
+ NT NT
110
5
+ 1:210
6
NT NT NT NT NT
110
6
1:210
7
NT NT NT NT NT
Vol ume 339 Number 12 789
the serum prostate-specific antigen concentration in
all eight patients; moreover, in the six patients who
stopped taking PC-SPES, prostate-specific antigen in-
creased within two to six weeks after treatment was
discontinued (Table 3). The magnitude of this effect
varied, perhaps because of the wide range of initial
prostate-specific antigen values or differences in the
types of local therapy the patients had received. The
increase in prostate-specific antigen after the discon-
tinuation of PC-SPES occurred at about the same
time as the increase in testosterone. For example, three
weeks after Patient 1 stopped taking PC-SPES, the
testosterone concentration had increased from 25 to
674 ng per deciliter (87 to 2337 nmol per liter) and
the prostate-specific antigen concentration had in-
creased from 0.4 ng per milliliter to 5.4 ng per milli-
liter, with an increase to 17.0 ng per milliliter at six
weeks.
Previous studies of patients with prostate cancer
have shown that the effects of pharmacologic doses
of estrogen are similar to those of PC-SPES.
16-19
Our
study assessed patients whose tumors were not known
to be refractory to androgen-ablation therapy; the ef-
fects of PC-SPES on cancer that is refractory to hor-
mone therapy and whether tumors would become re-
sistant to the effects of this compound are unknown.
20
The adverse effects of PC-SPES were similar to
those of estrogen. All patients in our study experi-
enced impotence and breast tenderness, and one pa-
tient had a superficial venous thrombosis. Prior studies
of estrogen therapy in men with prostate cancer have
assessed the frequency of these effects.
16-19
For exam-
ple, the Leuprolide Study Group reported that 49
percent of patients who were treated with diethylstil-
bestrol for metastatic prostate cancer experienced
breast tenderness, and 7 percent had venous throm-
bosis.
16
Aprikian et al. also found that the most com-
mon adverse effect of diethylstilbestrol treatment was
gynecomastia and that 2 of 55 patients had throm-
bosis (1 had a pulmonary embolus).
18
Our laboratory data support the hypothesis that
the clinical effects of PC-SPES are due at least in
part to estrogenic activity. PC-SPES activates the
human estrogen-response element in two yeast sys-
tems. The estrogenic activity of a 1:200 dilution of
a stock extract of one 320-mg PC-SPES capsule was
similar to that of 1 nM estradiol in both the yeast
lacZ and growth assays. PC-SPES also had estrogenic
activity in mice. Ovariectomized CD-1 mice given
PC-SPES had significantly heavier uteri than control
mice. Other groups have used this mouse model to
study the estrogenicity of various compounds.
15,21
The CD-1 mouse has also been used to study the ter-
atogenic effects of estrogens.
22,23
Prenatal exposure
of CD-1 mice to diethylstilbestrol results in repro-
ductive tract anomalies similar to those that occur in
women who were exposed to diethylstilbestrol pre-
natally.
23
Given the potential toxicity of PC-SPES in
pregnant women, studies of this herbal combination
in animal models are warranted.
*The references ranges for serum testosterone at four standardized commercial laboratories were
241 to 827, 220 to 940, 220 to 940, and 300 to 1000 ng per deciliter. To convert values for testos-
terone to nanomoles per liter, multiply by 3.467. NA denotes not applicable, since these patients did
not stop taking PC-SPES.
TABLE 3. EFFECT OF PC-SPES ON TESTOSTERONE AND PROSTATE-SPECIFIC ANTIGEN
CONCENTRATIONS.*
PATIENT
NO.
PRIOR LOCAL
THERAPY FOR
PROSTATE CANCER TESTOSTERONE PROSTATE-SPECIFIC ANTIGEN
INTERVAL BETWEEN
DISCONTINUATION
OF PC-SPES AND
MEASUREMENT
DURING
PC-SPES
AFTER
PC-SPES
BEFORE
PC-SPES
DURING
PC-SPES
AFTER
PC-SPES
ng/dl ng/ml wk
1 Prostatectomy 25 674 34.6 0.4 5.4
17.0
3
6
2 Radiation therapy 149 301 1.7 1.1 1.7 2
3 None 26 1072 122 1.2 36.0 2
4 Prostatectomy fol-
lowed by radiation
therapy
494 1324 1.2 0.5 0.8 2
5 Radiation therapy fol-
lowed by cryo-
surgery
362 986 1.3 0.4 0.8 2
6 None 363 918 12.6 6.3 7.1 2
7 None 53 NA 6.7 5.6 NA NA
8 Prostatectomy 316 NA 0.39 0.0 NA NA
790 September 17, 1998
0.0
1.2
0 60
0.4
0.8
10 20 30 40 50
Minutes
PC-SPES
DES
Peak Y
m/z
100%
75%
50%
25%
0%
40
73
207 254 281 355
385
147
T
o
t
a
l

I
o
n

I
n
t
e
n
s
i
t
y
0
15
5
10
m/z
100%
75%
50%
25%
0%
40
73
115 149 177
217
279
384
413
0.0
0.3
0 70
0.1
0.2
10 20 30 40 50 60
Minutes
PC-SPES
Estrogens
Estradiol
Estrone
DES
Peak X
O
p
t
i
c
a
l

D
e
n
s
i
t
y

a
t

2
5
4

n
m
0.0
0.3
0.1
0.2
B
D
C
A
Vol ume 339 Number 12 791
Halicka et al. found that PC-SPES inhibited the
growth of PC-3 cells (prostate-tumor cells) and MCF-7
cells (breast-tumor cells) in cell culture.
7
Hsieh et al.
demonstrated that PC-SPES decreased both the pro-
duction of prostate-specific antigen and the expres-
sion of androgen receptors in cultures of LNCaP
prostate-tumor cells.
8
We do not know whether
these effects of PC-SPES on cultured tumor cells are
independent of estrogenic activity, since estrogen
alone induces apoptosis in tumor-cell lines.
24
PC-SPES has potent estrogenic activity and con-
tains multiple organic compounds, but not the
known estrogens estrone, estradiol, and diethylstil-
bestrol or chemicals with similar structures and me-
tabolites. Since our laboratory found estrogenic ac-
tivity in elutants of PC-SPES corresponding to
separate peaks obtained from high-performance liq-
uid chromatography, multiple phytoestrogens may
be responsible for the observed activity. Our results
suggest that PC-SPES may prove useful in the treat-
ment of hormonally sensitive prostate cancer; but
when used concurrently with standard or experi-
mental therapies PC-SPES may confound the re-
sults. In addition, estrogens can have substantial tox-
ic effects, and the safety of nutritional supplements
with substantial estrogenic activity needs to be eval-
uated. These data demonstrate that unregulated,
commercially available dietary supplements may have
biologic activity that can affect diseases, standard
medical therapy, and general health.
Supported in part by grants from the National Cancer Institute (NCI-
CA72720, RO3-CA77133, and CA57142), Environmental and Occupa-
tional Health Sciences Institute (ES05022), and the Environmental Pro-
tection Agency (R825386-010).
We are indebted to L. Korn, Ph.D., Department of Environmen-
tal and Community Medicine and Center for Biostatistics, Robert
Wood Johnson Medical School, for biostatistical analysis and to Ted
Colterelli for collecting data on the patients.
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Figure 3. Results of High-Performance Liquid Chromatography,
Gas Chromatography, and Mass Spectrometry of PC-SPES and
the Estrogens Estrone, Estradiol, and Diethylstilbestrol (DES).
The results of high-performance liquid chromatography, shown
in Panels A and B, are representative of three experiments with
two separate batches of PC-SPES. The results of gas chromatog-
raphy and mass spectrometry show that peak X of PC-SPES in
Panel B differs from the peak representing diethylstilbestrol in
Panel A. Gas chromatograms of diethylstilbestrol (Panel C) and
peak X of PC-SPES (Panel D) are shown along with in-line mass
spectrometric analysis of diethylstilbestrol (inset in Panel C) and
peak Y of peak X (inset in Panel D). The mass spectrometer was
set to scan particles with a mass-to-charge ratio (m/z) ranging
from 17 to 600 every 0.6 second.

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