Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines
Aim: Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major cause...
Збережено в:
| Дата: | 2013 |
|---|---|
| Автори: | , , , , |
| Формат: | Стаття |
| Мова: | English |
| Опубліковано: |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2013
|
| Назва видання: | Experimental Oncology |
| Теми: | |
| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/139121 |
| Теги: |
Додати тег
Немає тегів, Будьте першим, хто поставить тег для цього запису!
|
| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines / M.W. Roomi, N.W. Roomi, B. Bhanap, A. Niedzwiecki, M. Rath // Experimental Oncology. — 2013. — Т. 35, № 1. — С. 20-24. — Бібліогр.: 10 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
irk-123456789-139121 |
|---|---|
| record_format |
dspace |
| spelling |
irk-123456789-1391212018-06-20T03:05:11Z Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines Roomi, M.W. Roomi, N.W. Bhanap, B. Niedzwiecki, A. Rath, M. Original contributions Aim: Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients. Methods: We investigated the effect of a nutrient mixture on Fanconi Anemia human fibroblast cell lines FA‐A:PD20 and FA‐A:PD220 on matrix metalloproteinase expression, invasion, cell proliferation, morphology and apoptosis. The cell lines were grown in a modified Dulbecco’s Eagle medium and at near confluence were treated with the nutrient mixture at increasing doses: 0; 10; 50; 100; 500; 1000 µg/ml. The cells were also treated with PMA to induce MMP-9 expression. Results: Zymography demonstrated MMP‐2 and PMA‐induced MMP‐9 activity. The nutrient mixture inhibited expression of both, MMP-2 and MMP-9, in a dose dependent manner with virtually total inhibition observed at 500 µg/ml. Matrigel invasion was inhibited in both cells lines; with 100% inhibition for FA-A:PD20 at 500 µg/ml and 100% inhibition of FA-A:P220 cells at 100 µg/ml. H&E staining did not indicate any change in cell morphology and causes apoptosis at higher doses. Conclusion: Our data demonstrated that the nutrient mixture inhibited matrix metalloproteinase expression, invasion and induced apoptosis, the important parameters for cancer prevention. The results suggest that the nutrient mixture may have therapeutic potential in Fanconi Anemia associated neoplasia. 2013 Article Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines / M.W. Roomi, N.W. Roomi, B. Bhanap, A. Niedzwiecki, M. Rath // Experimental Oncology. — 2013. — Т. 35, № 1. — С. 20-24. — Бібліогр.: 10 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/139121 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Original contributions Original contributions |
| spellingShingle |
Original contributions Original contributions Roomi, M.W. Roomi, N.W. Bhanap, B. Niedzwiecki, A. Rath, M. Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines Experimental Oncology |
| description |
Aim: Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients. Methods: We investigated the effect of a nutrient mixture on Fanconi Anemia human fibroblast cell lines FA‐A:PD20 and FA‐A:PD220 on matrix metalloproteinase expression, invasion, cell proliferation, morphology and apoptosis. The cell lines were grown in a modified Dulbecco’s Eagle medium and at near confluence were treated with the nutrient mixture at increasing doses: 0; 10; 50; 100; 500; 1000 µg/ml. The cells were also treated with PMA to induce MMP-9 expression. Results: Zymography demonstrated MMP‐2 and PMA‐induced MMP‐9 activity. The nutrient mixture inhibited expression of both, MMP-2 and MMP-9, in a dose dependent manner with virtually total inhibition observed at 500 µg/ml. Matrigel invasion was inhibited in both cells lines; with 100% inhibition for FA-A:PD20 at 500 µg/ml and 100% inhibition of FA-A:P220 cells at 100 µg/ml. H&E staining did not indicate any change in cell morphology and causes apoptosis at higher doses. Conclusion: Our data demonstrated that the nutrient mixture inhibited matrix metalloproteinase expression, invasion and induced apoptosis, the important parameters for cancer prevention. The results suggest that the nutrient mixture may have therapeutic potential in Fanconi Anemia associated neoplasia. |
| format |
Article |
| author |
Roomi, M.W. Roomi, N.W. Bhanap, B. Niedzwiecki, A. Rath, M. |
| author_facet |
Roomi, M.W. Roomi, N.W. Bhanap, B. Niedzwiecki, A. Rath, M. |
| author_sort |
Roomi, M.W. |
| title |
Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines |
| title_short |
Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines |
| title_full |
Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines |
| title_fullStr |
Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines |
| title_full_unstemmed |
Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines |
| title_sort |
repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2013 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/139121 |
| citation_txt |
Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines / M.W. Roomi, N.W. Roomi, B. Bhanap, A. Niedzwiecki, M. Rath // Experimental Oncology. — 2013. — Т. 35, № 1. — С. 20-24. — Бібліогр.: 10 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT roomimw repressionofmatrixmetalloproteinasesandinhibitionofcellinvasionbyanutrientmixturecontainingascorbicacidlysineprolineandgreenteaextractonhumanfanconianemiafibroblastcelllines AT roominw repressionofmatrixmetalloproteinasesandinhibitionofcellinvasionbyanutrientmixturecontainingascorbicacidlysineprolineandgreenteaextractonhumanfanconianemiafibroblastcelllines AT bhanapb repressionofmatrixmetalloproteinasesandinhibitionofcellinvasionbyanutrientmixturecontainingascorbicacidlysineprolineandgreenteaextractonhumanfanconianemiafibroblastcelllines AT niedzwieckia repressionofmatrixmetalloproteinasesandinhibitionofcellinvasionbyanutrientmixturecontainingascorbicacidlysineprolineandgreenteaextractonhumanfanconianemiafibroblastcelllines AT rathm repressionofmatrixmetalloproteinasesandinhibitionofcellinvasionbyanutrientmixturecontainingascorbicacidlysineprolineandgreenteaextractonhumanfanconianemiafibroblastcelllines |
| first_indexed |
2025-07-10T07:37:26Z |
| last_indexed |
2025-07-10T07:37:26Z |
| _version_ |
1837244669670457344 |
| fulltext |
20 Experimental Oncology 35, 20–24, 2013 (March)
REPRESSION OF MATRIX METALLOPROTEINASES AND INHIBITION
OF CELL INVASION BY A NUTRIENT MIXTURE, CONTAINING
ASCORBIC ACID, LYSINE, PROLINE, AND GREEN TEA EXTRACT
ON HUMAN FANCONI ANEMIA FIBROBLAST CELL LINES
M.W. Roomi, N.W. Roomi, B. Bhanap, A. Niedzwiecki*, M. Rath
Dr. Rath Research Institute, Cancer Division, Santa Clara, CA-95050, USA
Aim: Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, pro‐
gressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carci‐
noma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients. Me thods:
We investigated the effect of a nutrient mixture on Fanconi Anemia human fibroblast cell lines FA‐A:PD20 and FA‐A:PD220 on ma‐
trix metalloproteinase expression, invasion, cell proliferation, morphology and apoptosis. The cell lines were grown in a modified
Dulbecco’s Eagle medium and at near confluence were treated with the nutrient mixture at increasing doses: 0; 10; 50; 100; 500;
1000 µg/ml. The cells were also treated with PMA to induce MMP‐9 expression. Results: Zymography demonstrated MMP‐2 and
PMA‐induced MMP‐9 activity. The nutrient mixture inhibited expression of both, MMP‐2 and MMP‐9, in a dose dependent
manner with virtually total inhibition observed at 500 µg/ml. Matrigel invasion was inhibited in both cells lines; with 100% inhibition
for FA‐A:PD20 at 500 µg/ml and 100% inhibition of FA‐A:P220 cells at 100 µg/ml. H&E staining did not indicate any change
in cell morphology and causes apoptosis at higher doses. Conclusion: Our data demonstrated that the nutrient mixture inhibited
matrix metalloproteinase expression, invasion and induced apoptosis, the important parameters for cancer prevention. The results
suggest that the nutrient mixture may have therapeutic potential in Fanconi Anemia associated neoplasia.
Key Words: Fanconi Anemia, MMP, invasion, nutrients, apoptosis.
Fanconi Anemia (FA) is a rare autosomal reces-
sive disorder that includes a constellation of clinical
symptoms and carries a high risk of developing cancer.
It is estimated to affect 1 in 350,000 births. Although
FA can be seen across the population, Ashkenazi Jews
and Afrikaans in South Africa are more likely to be the
carriers of this chromosomal abnormality. Mean age
of diagnosis is 7 years [1] with the approximate age
at death of 35–40 years. Several common clinically ob-
served congenital defects include short stature, abnor-
malities of the skin, kidneys, ears and eyes. Additionally,
patients with FA may experience vertebral anomalies,
anal atresia, cardiac abnormalities, tracheo-esophageal
fistula, renal anomalies and radial limb, although 25%
patients may not have any physical abnormalities [2].
The hallmark of FA, however, is hematologic abnor-
mality including aplastic anemia and myelodysplastic
syndrome in childhood with a 90% probability of bone
marrow failure by 40 years [3]. Furthermore, patients
with FA have a 20% chance of developing cancers. The
highest chance is that of acute myeloid leukemia fol-
lowed by non-hematologic solid tumors as squamous
cell carcinomas of head and neck and ano-genital
regions. Other solid tumors include esophageal car-
cinoma, tumors of the liver, brain, skin and kidney [4].
We have developed strategies to inhibit cancer
development and its spread using naturally occurring
nutrients such as lysine, proline, ascorbic acid and
green tea extract. In our previous studies, this nutrient
mixture (NM) has exhibited a broad spectrum biologi-
cal activity, including synergistic effect and anticancer
activity, in vivo in a number of cancer cell lines through
inhibition of cancer cell growth, MMP secretion, inva-
sion, metastasis, and angiogenesis [5–7].
FA fibroblasts are different from normal fibroblasts,
because they have been shown to have impaired DNA
repair process. Additionally, the nuclear extracts have
substantially decreased plasmid-rejoining activity [8,
9]. This prompted us to study the effect of NM on FA fi-
broblast. In the current study, we investigated the ef-
fect of our NM on the proliferation, invasion, MMP
expression, morphology, and apoptosis of the human
FA fibroblast cell lines FA-A:PD20 and FA-A:PD220.
MATERIALS AND METHODS
NM. The composition of the NM and the proportion
included the following: vitamin C (as ascorbic acid and
as Mg, Ca and palmitate ascorbate) 710 mg; L-lysine
1,000 mg; L-proline 750 mg; L-arginine 500 mg; N-
acetyl cysteine 200 mg; standardized green tea extract
(80% polyphenol) 1,000 mg; selenium 30 µg; copper
2 mg; and manganese 1 mg.
Cell line and culture. The FA fibroblast cell lines
FA-A:PD220 and FA-A:PD20 were obtained from
Fanconi Anemia Research Fund (Eugene, OR, USA).
The cells were grown in modified Dulbecco’s Eagle
medium supplemented with 10% fetal bovine serum,
100 U/ml penicillin and 100 µg/ml streptomycin (Gibco
BRL, Long Island, NY) in 24-well tissue culture plates
(Costar, Cambridge, MA). Cells were incubated with
1 ml of media at 37 °C in a tissue culture incubator
Received: August 30, 2012.
*Correspondence: Fax: 408‑567‑5030
E‑mail: author@drrath.com
Abbreviations used: FA — Fanconi anemia; MMP — matrix metal‑
loproteinase; NM — nutrient mixture; PMA — phorbol 12‑myristate
13‑acetate.
Exp Oncol 2013
35, 1, 20–24
Experimental Oncology 35, 20–24, 2013 (March)35, 20–24, 2013 (March) (March) 21
equilibrated with 95% air and 5% CO. At near conflu-
ence, the wells were treated with the NM, dissolved
in the media and tested with the NM at the following
doses in triplicate: 0, 10, 50,100, 500 and 1,000 µg/ml.
Cells were also treated with PMA (100 ng/ml) to induce
MMP-9 secretion. The plates were then returned to the
incubator.
MTT cell proliferation assay. Cell viability was
evaluated by MTT assay, as described in the reference
[5, 6]. In brief, after 24 h incubation, the cells were
washed with phosphate buffered saline (PBS) and
500 µl of MTT (Sigma#M-2128) 0.5 mg/ml in media
was added. Following 12 h incubation, the supernatant
was carefully removed from the wells, the formazan
product was dissolved in 1 ml of DMSO, and absor-
bance was measured at 570 nm in Bio Spec 1601,
Shimadzu spectrometer. The OD570 of the DMSO
solution in each well was considered to be proportional
to the number of cells. The OD of the control (treatment
without supplement) was considered 100%.
Gelatinase zymography. Gelatinase zymography
was performed as described in the reference [6, 7].
In brief, the culture media (20 µl) were mixed with
sample buffer and loaded for SDS polyacrylamide gel
electrophoresis (SDS-PAGE) with tris-glycine SDS buf-
fer. Samples were not boiled before electrophoresis.
Following electrophoresis the gels were washed twice
in 2.5% Triton X-100 for 30 min at room tempera-
ture to remove SDS. The gels were then incubated
at 37 °C overnight in CaCl2 at pH 8.0 and stained with
0.5% Coomassie Blue R250 in 50% methanol and
10% glacial acetic acid for 30 min and de-stained;
producing clear bands against an intensely stained
background. Gelatinase zymograms were scanned
using CanoScan 9950F Canon scanner at 1,200 dpi.
The intensity of the bands was evaluated using a pixel-
based densitometer program Un-Scan-It, Version 5.1,
32-bit (Silk Scientific Corporation), at a resolution
of 1 Scanner Unit (1/100 dpi), and expressed as a per-
centage of control.
Matrigel invasion assay. Invasion studies were
conducted as described in the reference [6, 7]. In brief,
FA-A:PD220 and FA-A:PD20 cells, suspended in me-
dium, were supplemented with the NM and seeded
on the insert in the well. Thus, both the medium on the
insert and in the well contained the same supplements.
The plates with the inserts were then incubated in a cul-
ture incubator equilibrated with 95% room air and 5%
CO for 24 h. After incubation, the media from the wells
were drawn. The cells on the upper surface of the in-
serts were gently scrubbed away with cotton swabs.
The cells that had penetrated the Matrigel membrane
and migrated onto the lower surface of the Matrigel
were stained with hematoxylin and eosin (H&E).
Morphology and apoptosis. For apoptosis stu-
dies, FA-A:PD220 and FA-A:PD20 cells, at near con-
fluence, were challenged with NM dissolved in media
at the experimental doses and incubated for 24 h.
The cell culture was washed with PBS and treated
with the caspase reagent as specified in the manu-
facturer’s protocol (Molecular Probes Image-IT Live
Green Poly Caspases Detection Kit135104, Invitrogen).
The cells were photographed under a fluorescence
microscope and counted. Green-colored cells repre-
sent viable cells, while yellow orange represents early
apoptosis and red represents late apoptosis.
Statistical analysis. The results were expressed
as mean ±S.D. Data was analyzed by independent t-test.
RESULTS
Cell proliferation. NM had no effect on cell pro-
liferation in the both cell lines compared to control
(data not shown).
Gelatinase zymography. Zymography demon-
strated MMP-2 and MMP-9 expression by untreated
and PMA treated FA-A:PD20 and FA-A: PD220 cells.
Fig. 1 a shows the effect of the NM on the untreated
cell line FA-A:PD20 and Fig. 1 b, after PMA induction.
The NM inhibited expression of both MMPs in a dose
dependent fashion with a total virtual inhibition observed
at 500 µg/ml. This was further confirmed by densitom-
etry analysis (Fig. 1 c, d). Similarly, the FA-A:PD220 cell
line (Fig. 2 a, b), both untreated and PMA treated cells
exhibited a dose-dependent inhibition of MMP-2 and
MMP-9 with virtually total inhibition at 500 µg/ml.
0
20
40
60
80
100
120
140
Control 10 50 100 500 1000
MMP-2
µg/ml
µg/ml
0
20
40
60
80
100
120
Control 10 50 100 500 1000
MMP-2
MMP-9
MMP-2 —
MMP-9 —
MMP-2 —
b
a
c
Fig. 1. The effect of the NM on MMP expression by human FA fi-
broblasts FA-A:PD20. Lane 1 — markers; Lane 2 — Control; Lane
3–7 — 10; 50; 100; 500 and 1,000 µg/ml, respectively. The gel
on the right (b) was treated with PMA (100 ng/ml) for MMP-9 in-
duction. 1 c, d — Densitometry analyses of dose dependent
decrease of MMP-2 and MMP-9 expression by FA-A:PD20 cells
without and with PMA treatment
22 Experimental Oncology 35, 20–24, 2013 (March)
MMP-9 —
— MMP-2 —
ba
1 2 3 4 5 6 7 1 2 3 4 5 6 7
Fig. 2. The effect of the NM on MMP expression by human
FA fibroblasts FA-A:PD220. Lane 1 — markers; Lane 2 — Con-
trol; Lane 3–7 — 10; 50; 100; 500 and 1,000 µg/ml, respectively.
a — without PMA, b — with PMA treatment
Matrigel invasion. Fig. 3 reveals a significant
dose-dependent inhibition of FA-A:PD20 cell invasion
through the Matrigel membrane. For FA-A:PD20 cells,
50; 80 and 100% inhibition was observed at 50,
100 and 500 µg/ml respectively (Fig. 3 c, d). All the
invasion photomicrographs support the extent of inhi-
bition. Similar results were obtained for FA-A:PD220.
(p value was <0.01 in both types of cells.)
Morphology and apoptosis. The analysis with live
green caspase, revealed a dose-dependent increase
in apoptosis at higher doses for the FA-A:PD220 cell
line, as seen in Fig. 4 a-e. Quantitative analysis of these
data (Fig. 4 f) showed a 16% rate of apoptosis starting
at 250 µg/ml (10% early, 6% late) followed by 48%
at 500 µg/ml (10% early and 38% late) and an 87%
rate of apoptosis at 1,000 µg/ml (23% early, 64% late).
A similar dose-dependent apoptotic pattern was seen
in FA-A:PD20 cell line (data not shown).
DISCUSSION
The results of these in vitro studies of human
FA fibroblast cancer cell lines FA-A:PD220 and FA-
A:PD20 suggest that the NM is effective in inhibiting
invasion through an ECM-like matrix. Indeed a robust
decrease in Matrigel invasion was observed with maxi-
mal effect observed at high doses for FA-A:PD220 and
FA-A:PD20 (100 and 500 µg/ml, respectively). Fur-
thermore, a dose-dependent decrease of MMP-2 and
MMP-9 expression was observed in both cell lines,
with virtual total inhibition at 500µg/ml. In addition, the
NM also induced apoptosis at higher doses.
Live Green Caspase analysis demonstrated signifi-
cant apoptosis in NM-treated FA fibroblasts. Repres-
sion in the MMP expression and inhibition through
Matrigel could possibly be due to pharmacological
effect of the NM.
ba
d
e
c
0
20
40
60
80
100
120
Control 50 100 500
µg/ml
Fig. 3. Effect of NM on Matrigel invasion of FA-A:PD20 cells. a–e: Photomicrographs and analytical representation of FA-A:PD20 cells
at varying NM concentrations (a — Control; b, c, d — 50; 100; 500 µg/ml respectively; e — Analytical representation, p<0.01)
Experimental Oncology 35, 20–24, 2013 (March)35, 20–24, 2013 (March) (March) 23
Matrix metalloproteinases (MMPs) are a fam-
ily of zinc-dependent proteolytic enzymes that able
to degrade connective tissue such as the basement
membrane collagen and are associated with cancer
metastasis and tumor angiogenesis. Two key gelatinases
within the MMP family, MMP-2 and MMP-9, play a key
role in the degradation of collagen type IV and gelatin,
important components of the extracellular matrix (ECM).
As such, high expression of these MMPs may play a role
in the malignancy potential of FA-associated cancers.
The NM showed significant inhibition of MMP-2 and
MMP-9 expression, important mediators of angiogenesis
and metastasis. Extra cellular matrix stability plays a key
role in the development of cancer, and lysine is a natural
inhibitor of proteolysis of the ECM [10]. Indeed, we have
previously shown the NM has inhibited cell invasion and
MMP production in multiple cancer cell lines, which sug-
gests an important role of the NM in cancer therapeutics.
The NM was formulated by defining the critical physi-
ological targets, such as ECM integrity and MMP activity,
in cancer progression and metastasis. The NM shows
activity against MMP expression, Matrigel invasion and
induces apoptosis at higher doses in human FA fibroblast
cell lines. These data suggest a role of NM in the possible
prevention and treatment of FA-associated solid tumors,
specifically by targeting MMP secretion and thereby
0
20
40
60
80
100
120
Control 100 250 500 1000
Live cells
Early apoptotic cells
Late apoptotic cells
µg/ml
ba
c d
e f
Fig. 4. a–e: Photomicrographs of FA-A:PD220 showing increasing apoptosis with increasing NM concentrations using live green
caspase (a — Control; b, c, d, e — 100; 250; 500; 1000 µg/ml respectively). f: Quantitative analysis of the cell line FA-A:PD220 show-
ing the increasing rate of apoptosis relative to control conditions
24 Experimental Oncology 35, 20–24, 2013 (March)
inhibiting migration of cancer within the ECM as well
as stabilizing the ECM surrounding an encapsulated
tumor. Overall, the NM may offer a therapeutic benefit
in FA associated neoplasia, and could play a role in sup-
port of FA patients.
ACKNOWLEDGEMENTS
Dr. Rath Health Foundation, a not for profit organi-
zation, provided research funding.
REFERENCES
1. Moustacchi E. Fanconi’s Anemia. Orphanet
En cyclop edia . O ct 2003 ht tp ://www.orpha.n et/
data/patho/Pro/en/FanconiAnemia-FRenPro634.pdf
2. D’Andrea AD. Susceptibility pathways in Fanconi’s ane-
mia and breast cancer. N Engl J Med 2010; 362: 1909–19.
3. Alter BP. Diagnosis, genetics, and management of in-
herited bone marrow failure syndromes. Hematology Am Soc
Hematol Educ Program, 2007; 29–39.
4. Auerbach AD. Fanconi anemia and its diagnosis. Mutat
Res 2007; 668: 4–10.
5. Roomi MW, Roomi N, Ivanov V, et al. Inhibitory ef-
fect of a mixture containing ascorbic acid, lysine, proline, and
green tea extract on critical parameters in angiogenesis. Oncol
Reports 2005, 14: 807–15.
6. Roomi MW, Roomi NW, Kalinovsky T, et al. Marked
inhibition of growth and invasive parameters of head and neck
squamos carcinoma FaDu by a nutrient mixture. Integrative
Cancer Ther 2009; 8: 168–76.
7. 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: 529–42.
8. Donahue SL, Lundberg R, Saplis R, et al. Deficient
regulation of DNA double-strand break repair in Fanconi
anemia fibroblasts. J Biol Chem 2003; 278: 29487–95.
9. Donahue SL, Campbell C. A DNA double strand break
repair defect in Fanconi anemia fibroblasts. J Biol Chem 2002;
277: 46243–7.
10. Rath M, Pauling L. Plasmin induced proteolysis and
the role of apoprotein (a), lysine and synthetic analogs. Or-
thomolecular Med 1992, 7: 17–23.
Copyright © Experimental Oncology, 2013
|