A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice
Background: Brain tumors are highly aggressive tumors characterized by secretions of high levels of matrix metalloproteinase-2 and -9, leading to tumor growth, invasion and metastasis by digesting the basement membrane and extracellular matrix components. We previously demonstrated the effectiveness...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Цитувати: | A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice / M.W. Roomi, T. Kalinovsky, M. Rath, A. Niedzwiecki // Experimental Oncology. — 2016 — Т. 38, № 1. — С. 54–56. — Бібліогр.: 9 назв. — англ. |
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irk-123456789-1379802018-06-18T03:02:47Z A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice Roomi, M.W. Kalinovsky, T. Rath, M. Niedzwiecki, A. Short communications Background: Brain tumors are highly aggressive tumors characterized by secretions of high levels of matrix metalloproteinase-2 and -9, leading to tumor growth, invasion and metastasis by digesting the basement membrane and extracellular matrix components. We previously demonstrated the effectiveness of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract in vitro: on activity of urokinase plasminogen activator, matrix metalloproteinases and TIMPs in various human glioblastoma (LN-18, T-98G and A-172) cell lines and on glioblastoma A-172 cell proliferation and Matrigel invasion. Aim: Our main objective in this study was to investigate the effect of the NM in vivo on human glioblastoma U-87 MG cell line. Materials and Methods: Athymic male nude mice inoculated with 3·106 U-87 MG cells subcutaneously and were fed a regular diet or a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed, the tumors were weighed and measured. The samples were studied histologically. Results: NM inhibited tumor weight and tumor burden by 53% (p = 0.015) and 48% (p = 0.010), respectively. Conclusions: These results suggest the therapeutic potential of NM as an adjuvant in the treatment of glioblastoma. 2016 Article A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice / M.W. Roomi, T. Kalinovsky, M. Rath, A. Niedzwiecki // Experimental Oncology. — 2016 — Т. 38, № 1. — С. 54–56. — Бібліогр.: 9 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/137980 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Short communications Short communications |
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Short communications Short communications Roomi, M.W. Kalinovsky, T. Rath, M. Niedzwiecki, A. A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice Experimental Oncology |
| description |
Background: Brain tumors are highly aggressive tumors characterized by secretions of high levels of matrix metalloproteinase-2 and -9, leading to tumor growth, invasion and metastasis by digesting the basement membrane and extracellular matrix components. We previously demonstrated the effectiveness of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract in vitro: on activity of urokinase plasminogen activator, matrix metalloproteinases and TIMPs in various human glioblastoma (LN-18, T-98G and A-172) cell lines and on glioblastoma A-172 cell proliferation and Matrigel invasion. Aim: Our main objective in this study was to investigate the effect of the NM in vivo on human glioblastoma U-87 MG cell line. Materials and Methods: Athymic male nude mice inoculated with 3·106 U-87 MG cells subcutaneously and were fed a regular diet or a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed, the tumors were weighed and measured. The samples were studied histologically. Results: NM inhibited tumor weight and tumor burden by 53% (p = 0.015) and 48% (p = 0.010), respectively. Conclusions: These results suggest the therapeutic potential of NM as an adjuvant in the treatment of glioblastoma. |
| format |
Article |
| author |
Roomi, M.W. Kalinovsky, T. Rath, M. Niedzwiecki, A. |
| author_facet |
Roomi, M.W. Kalinovsky, T. Rath, M. Niedzwiecki, A. |
| author_sort |
Roomi, M.W. |
| title |
A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice |
| title_short |
A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice |
| title_full |
A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice |
| title_fullStr |
A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice |
| title_full_unstemmed |
A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice |
| title_sort |
nutrient mixture inhibits glioblastoma xenograft u-87 mg growth in male nude mice |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2016 |
| topic_facet |
Short communications |
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http://dspace.nbuv.gov.ua/handle/123456789/137980 |
| citation_txt |
A nutrient mixture inhibits glioblastoma xenograft U-87 MG growth in male nude mice / M.W. Roomi, T. Kalinovsky, M. Rath, A. Niedzwiecki // Experimental Oncology. — 2016 — Т. 38, № 1. — С. 54–56. — Бібліогр.: 9 назв. — англ. |
| series |
Experimental Oncology |
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2025-07-10T02:41:48Z |
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2025-07-10T02:41:48Z |
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| fulltext |
54 Experimental Oncology 38, 54–56, 2016 (March)
A NUTRIENT MIXTURE INHIBITS GLIOBLASTOMA XENOGRAFT
U-87 MG GROWTH IN MALE NUDE MICE
M.W. Roomi, T. Kalinovsky, M. Rath, A. Niedzwiecki*
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050, USA
Background: Brain tumors are highly aggressive tumors characterized by secretions of high levels of matrix metalloproteinase-2 and
-9, leading to tumor growth, invasion and metastasis by digesting the basement membrane and extracellular matrix components.
We previously demonstrated the effectiveness of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea
extract in vitro: on activity of urokinase plasminogen activator, matrix metalloproteinases and TIMPs in various human glioblas-
toma (LN-18, T-98G and A-172) cell lines and on glioblastoma A-172 cell proliferation and Matrigel invasion. Aim: Our main
objective in this study was to investigate the effect of the NM in vivo on human glioblastoma U-87 MG cell line. Materials and
Methods: Athymic male nude mice inoculated with 3·106 U-87 MG cells subcutaneously and were fed a regular diet or a regular
diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed, the tumors were weighed and measured. The samples
were studied histologically. Results: NM inhibited tumor weight and tumor burden by 53% (p = 0.015) and 48% (p = 0.010), re-
spectively. Conclusions: These results suggest the therapeutic potential of NM as an adjuvant in the treatment of glioblastoma.
Key Words: glioblastoma U-87 MG cell line, nutrients, xenograft tumor growth.
Approximately 22,850 new cases and 15,320 deaths
from brain and nervous system tumors were estimated
in the United States for adults in 2015 [1]. Brain cancer
(mainly gliomas and medulloblastomas) is the second
(after leukemia) leading cause of cancer death in chil-
dren aged under 20, accounting for one third of all
cancer deaths in this age group [2]. Though most brain
tumors tend not to metastasize to distant areas outside
the brain and/or spinal cord, they do tend to recur lo-
cally, or spread to other areas of the central nervous
system. Tumor cell invasion depends upon degrada-
tion of the extracellular matrix, which, when intact, acts
as a barrier to block cancer cell invasion [3, 4]. Rath
and Pauling proposed a universal approach to tumor
growth and expansion using nutrients such as lysine
and ascorbic acid to target plasmin-mediated con-
nective tissue degradation [5]. Subsequent studies
confirmed this approach and led to identifying a novel
formulation composed of lysine, ascorbic acid, pro-
line and green tea extract and other micronutrients,
which has shown significant anticancer activity against
a large number (~40) of cancer cell lines, blocking
cancer growth, tissue invasion and matrix metallopro-
teinase (MMP) expression both in vitro and in vivo [6].
In a previous in vitro study, the nutrient mixture
(NM) significantly inhibited urokinase plasminogen
activator (uPA) and MMP secretion and increased
TIMP-2 secretion in glioma cell lines [7]. Other stu-
dies found significant inhibition of cell proliferation and
Matrigel invasion of glioma cell line A-172 by NM [8]
and significant negative correlation between NM inhi-
bition of Matrigel invasion and NM modulation of the
MMP-2 activity of glioma cell line A-172 [9]. Our main
objective in this study was to investigate the effect
of NM in vivo using human glioblastoma U-87 MG xe-
nografts in male nude mice.
MATERIALS AND METHODS
Cell line and NM. Human glioblastoma cell line
U-87 MG was obtained from ATCC (American Type
Culture Collection, Rockville, MD, USA). The NM, pre-
pared by VitaTech (Hayward, CA), was composed of the
following ingredients in the amounts indicated: Vitamin
C (as ascorbic acid and as Mg, Ca, and palmitate ascor-
bate) 700 mg; L-lysine 1000 mg; L-proline 750 mg;
L-arginine 500 mg; N-acetyl cysteine 200 mg; stan-
dardized green tea extract (80% polyphenol) 1000 mg;
selenium 30 μg; copper 2 mg; manganese 1 mg.
Animals. Male nude mice, approximately 5 weeks
of age on arrival, were purchased from Simonsen
Labo ratories, Gilroy, CA, and maintained in microiso-
lator cages under pathogen-free conditions on a 12-
hour light/12-hour dark schedule for a week. All pro-
cedures were performed according to humane and
customary care and use of experimental animals and
followed a protocol approved by internal institutional
animal safety review committee.
Experimental design. After housing for a week,
the mice (n = 12) were inoculated subcutaneously
with 3•106 glioblastoma U-87 MG cells in 0.2 ml PBS
and 0.1 ml Matrigel (BD Bioscience, Bedford, MA).
After injection, the mice were randomly divided into
two groups; the control group of mice was fed regular
Laboratory Rodent Diet 5001 from Purina Mills, LLC
(Gray Summit, MO, USA). and the NM group — the
regular diet supplemented with 0.5% NM (w/w). The
0.5% NM diet was milled and pressed by Purina Mills,
LLC and generated by Vitatech (Tustin, CA, USA).
During the study, the mice consumed, on the average,
4 g of their respective diets per day. Thus, the supple-
mented mice received approximately 20 mg of NM per
day. After four weeks, the mice were sacrificed and
their tumors were excised and processed for histo logy.
Dimensions (length and width) of tumors were mea-
Submitted: December 8, 2015.
*Correspondence: E-mail: author@drrath.com
Abbreviations used: MMP — matrix metalloproteinase; NM — nutri-
ent mixture; uPA — urokinase plasminogen activator.
Exp Oncol 2016
38, 1, 54–56
SHORT COMMUNICATIONS
Experimental Oncology 38, 54–56, 2016 (March) 55
sured using a digital caliper, and the tumor burden was
calculated using the following formula: 0.5 × length ×
width. Mean weight of mice at initiation of study and
termination of study did not differ significantly between
the groups.
Histology. Tissue samples were fixed in 10% buf-
fered formalin. All tissues were embedded in paraffin
and cut at 4–5 microns. Sections were deparaffinized
through xylene and graduated alcohol series to water
and stained with hematoxylin and eosin (H & E) for
evaluation using a standard light microscope.
Statistical analysis. The results were expressed
as means ± SD and analyzed by independent sample
“t” test using MedCalc software (Mariakerke, Bel-
gium).
RESULTS AND DISCUSSION
NM strongly inhibited the growth of U-87 MG xe-
nografts in male nude mice. Mean tumor weight was
inhibited by 53% (p = 0.0147) with NM 0.5% dietary
supplementation, as shown in Fig. 1, a and tumor
burden was inhibited by 48% (p = 0.0098), as shown
in Fig. 1, b. There was no significant difference be-
tween mean body weights of groups of mice.
0
0.5
1
1.5
2
2.5
3
3.5
Control NM 0.5%
*
M
ea
n
tu
m
or
w
ei
gh
t,
g
0
100
200
300
400
500
Control NM 0.5%
M
ea
n
tu
m
or
b
ur
de
n,
m
m
2
*
ba
Fig. 1. Effect of NM on mean tumor weight (a) and tumor burden
(b) in nude mice grafted with U-87 MG cells. *p = 0.015 (a) and
p = 0.01 (b) as compared to control
Histologically, the tumors from both groups were
irregularly round, subcutaneous masses, composed
of solid sheets and nests of irregularly round epithe-
lioid cells with prominent round to ovoid nuclei and
lightly staining bluish cytoplasm with ill-defined cell
borders. Tumors from control and NM-supplemented
mice were similar morphologically, but the tumors from
supplemented mice were significantly smaller in size.
Areas of tumor necrosis ranged from 50–80% of tu-
mor mass in the control group compared to 10–60%
in the NM group. Areas of tumor necrosis were associ-
ated with moderate to severe neutrophilic infiltration
(Fig. 2, a–d).
Previous in vitro results from the cellular proliferation
study of glioblastoma A-172 support the in vivo find-
ings, as NM showed increased toxicity in cells in a dose-
dependent manner, with 50% inhibition of cell growth
in cells exposed to 1000 μg/ml NM and total block
of invasion through Matrigel at 1000 μg/ml NM [8].
In addition, prior studies of seve ral glioblastoma cell
lines, demonstrated potent, significant suppression
of invasive parameters by the NM, including uPA and
MMP-2 and -9 [7].
Brain cancer is the second leading cause of cancer
death in children under 20 years, accounting for one
third of all cancer deaths in this age group. Current
treatment methods for gliomas are generally ineffec-
tive and toxic. Thus, there is a need for development
of effective therapeutic agents for these cancers with
minimal toxicity. Our studies demonstrated that the
mixture of the non-toxic components of NM signifi-
cantly inhibited the growth and tumor burden of glio-
blastoma cell line U-87 MG in vivo. These results
in addition to our prior in vitro studies with glioblastoma
cell lines, which documented significant inhibition
of MMP-2 and-9 secretion and invasion by NM, sug-
gest the potential of NM as an adjuvant in treatment
of glioblastomas.
ba
c d
Fig. 2. Histopathology of tumors from control (a, b) and NM-
supplemented (c, d) mice, × 200. In b and d tumor necrosis and
associated neutrophilic inflammation are present
ACkNOWLEDGEMENTS
The research study was funded by Dr. Rath Health
Foundation (Santa Clara, CA, USA), a non-profit orga-
nization. We thank consulting pathologist Alexander
de Paoli, DVM, PhD, IDEXX Reference Laboratories
for providing histopathology slides.
REFERENCES
1. ACS. What are the key statistics about brain and spinal cord
tumors? (http://www.cancer.org/cancer/braincnstumorsinadults/
detailedguide/brain-and-spinal-cord-tumors-in-adults-key-
statistics).
2. ABTA (American Brain Tumor Association). Brain tumor facts
(http://www.abta.org/about-us/news/brain-tumor-statistics/).
3. Yurchenko PD, Schitny JC. Molecular architecture
of basement membranes. FASEB J 1990; 4: 1577–90.
56 Experimental Oncology 38, 54–56, 2016 (March)
4. Barsky SH, Siegel GP, Jannotta F, et al. Loss of base-
ment membrane components by invasive tumors but not
by their benign counterparts. Lab Investig 1983; 49: 140–7.
5. Rath M, Pauling L. Plasmin-induced proteolysis and the
role of apoprotein(a), lysine and synthetic analogs. Orthomol
Med 1992; 7: 17–23.
6. Niedzwiecki A, Roomi MW, Kalinovsky T, et al. Micro-
nutrient synergy — a new tool in effective control of metastasis
and other key mechanisms of cancer. Cancer Metastasis Rev
2010; 29: 529–43.
7. Roomi MW, Kalinovsky T, Niedzwiecki A, et al. Modu-
lation of uPA, MMPs and their inhibitors by a novel nutrient
mixture in human glioblastoma cell lines. Int J Oncol 2014;
45: 887–94.
8. Roomi MW, Ivanov V, Kalinovsky T, et al. Inhibition
of glioma cell line A-172 MMP activity and cell invasion in vitro
by a nutrient mixture. Med Oncol 2007; 24: 231–8.
9. Roomi MW, Monterrey JC, Kalinovsky T, et al. Inhibition
of invasion and MMPs by a nutrient mixture in human cancer
cell lines: a correlation study. Exp Oncol 2010; 32: 243–8.
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