| Suppression
of MMP Expression and Invasion of Human Cervical Cancer Cell lines
Hela and DoTc2 4510 by Nutrients
M.W. Roomi, V. Ivanov, A. Niedzwiecki, M. Rath
Presented at: 44th Annual Meeting of the Society
of Toxicology, New Orleans, March 6-10, 2005.
Published in: Proceedings of the 44th Annual
Meeting of the Society of Toxicology, Abstract #1494
Abstract
Background:
Cervical cancer is the seventh most common cancer worldwide and
the second most common cancer in women. Early detection by screening
followed by appropriate treatment is associated with a high cure
and survival rate. Untreated dysplasia and carcinoma in situ leads
to 30% to 40% invasion within ten years.
Objective:
We investigated the synergistic effect of a unique nutrient formulation
(NM) containing lysine, proline, arginine, ascorbic acid, and
epigallocatechin gallate on human cervical cancer cells HeLa (adenocarcinoma
of cervix - CCL2) and DoTc2 4510 (carcinoma of cervix - CRL7920)
by measuring: cell proliferation, modulation of MMP-2 and –9
expression, and cancer cell invasive potential.
Materials and Methods:
Human cervical Hela (CCL-2) and carcinoma DoTc2 4510 (CRL 7920),
obtained from ATCC, were grown in DME medium supplemented with
10% FBS, penicillin (100 U/ml) and streptomycin (100 mg/ml) in
24-well tissue culture plates. At near confluence, the cells were
treated with the nutrient mixture (NM) dissolved in media and
tested at 0, 10, 100, 500, and 1000 ?g/ml in triplicate at each
dose. Cells were also treated with phorbyl myrsitate (PMA ) 200
ng/ml to study enhanced expression of MMP-9. Cell proliferation
was evaluated by MTT assay, MMP expression by gelatinase zymography,
and invasion through Matrigel.
Results:
NM with and without PMA 200 ng/ml showed significant antiproliferative
effect on human cervical CCL-2 cancer cell growth (untreated 37%
treated 57% at 1000 µg/ml)(p<0.0002). NM significantly
inhibited cervical DoTc2 4510 cancer cell growth (untreated 45%
treated 49%)(p<0.0004). Zymography demonstrated expression
of MMP-2 by untreated cervical CCL-2 and enhanced MMP-2 and induced
MMP-9 by PMA (200 ng/ml) treated CCL-2 cells. NM inhibited the
CCL-2 expression of MMP-2 and –9 in a dose-dependent fashion,
with virtual total inhibition of MMP-2 at 1000 µg/ml and
MMP-9 at 500 µg/ml concentration. Untreated DoTc2 4510 cells
demonstrated MMP-9 expression, which was enhanced with PMA treatment.
NM inhibited MM-9 expression in a dose-dependent fashion with
virtual inhibition at 500 µg/ml. The synergistically acting
nutrient mixture significantly reduced the invasion of human cervical
cancer cells CCL-2 through Matrigel in a dose-dependent fashion,
with 76% inhibition at 100 µg/ml and 100% at 500 µg/ml
NM (p<0.0001). Invasion inhibition of cervical cancer cells
DoTc2 4510 97% at 500 µg/ml and 100% at 1000 µg/ml
(p<0.0001).
Conclusion:
Our results suggest that NM is an excellent candidate for therapeutic
use in the treatment of cervical cancer, by inhibiting critical
steps in cancer development and spread, such as cell growth, MMP
expression and invasion.
Comment:
Cervical cancer is the seventh most common cancer worldwide
and the second most common cancer in women. Untreated dysplasia
and carcinoma in situ leads to 30% to 40% invasion within
ten years. We studied the synergistic effect of a unique nutrient
formulation containing lysine, proline, arginine, ascorbic
acid, and epigallocatechin gallate on metastatic parameters
on human cervical cancer cells such as effect on MMP-2 and
–9 expression and cancer cell invasive potential. Our
results demonstrated complete inhibition of Hela MMP-2 expression
at 1000 µg/ml and MMP-9 at 500 µg/ml, and of Matrigel
invasion at 500 ?g/ml. NM demonstrated complete inhibition
of cervical DoTc2 4510 cell expression of MMP-9 at 500 µg/ml
and of invasion at 1000 µg/ml. These findings are significant
as they suggest that the nutrient mixture would be an effective
and safe therapeutic regimen for treatment of cervical cancer.
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