Biology and biotech-

nology
The Effect of Methanolic Sabal palmetto Extract
on MCF7 Breast Cancer Cells

Dana Ahmad 1910257

Jibreel Ayyad 1910015
Sara Al Tamam 1910186
Introduction

 Cancer is a serious issue and a condition

that can be fatal.
• Terms: metastasis/apoptosis/
angiogenesis/MCF7 cells.
 Sabal palmetto identification.
• Common name
• Origin
• Genus, species
 IntroductionCont.
Aim of the study: to study the ef -
fect of methanolic extract of S.-
palmetto on MCF7 breast cancer
cells.
Results:
• Low cytotoxicity
• High angiogenesis
• Apoptotic induction.
Methodology
1.Preparation of plant extract.
• The stems of S. palmetto were gathered, cleaned, dried, and then coarsely grounded,
Refluxing with methanol solvent, filtered, then evaporated until it was completely dry.

• Dimethyl sulphoxide (DMSO) was used to dissolve 1 g of the crude extract.

• The extract underwent filtration, and were stored at -20 °C.

 Methodology

2.Preparation of MCF7 cells.

• The adherent human breast cancer cell line MCF7 was grown in RPMI 1640 or
Roswell Park Memorial Institute Medium (RPMI), pH 7.4, aided with 10% fetal
bovine serum (FBS), gentamicin, 2-mercaptoethanol, N-2 hydroxyethylpiperzine-
N-2-ethane sulfonic acid (HEPES), sodium pyruvate, penicillin, streptomycin,
and L-glutamine, and it was harvested at 70% confluence and sub-cultured.

• Trypsinized cells were reseeded in fresh media at a density of approximately

105 cells per milliliter and kept at 37°C in a humidified 5% CO2 incubator.
 Methodology
3. In-vitro cytotoxicity assay
• MTT test method was employed to evaluate the degree of cell viability.

• In 96-well plates with cells per well and media, cells (MCF7) were cultivated. They were then incubated
for 24 hours. Then, a fresh medium and 100 µl of medium containing S. palmetto plant extracts at
various concentrations (12.5, 25, 50, 100, 200, and 400 µg/ml) were added to the cells on the plates.

• Parallel to each test, a negative control was run in triplicate using all vehicle components other than plant
extract.

• The medium was removed from each well and 20 ul/well of mTT dye solution diluted in PBS was added.

• Plates were incubated for 4 hours. After that, each well received 100 µl of DMSO, which was properly
mixed in to dissolve the dye crystals before it was immediately incubated for 15 minutes at room
temperature in the dark.
 Methodol-
ogy
3. In-vitro cytotoxicity assay
 Methodology

4. Cell migration assay ( wound-healing technique).

• A 6-well plate was seeded with a significant number of MCF7 cells and left to incubating overnight.

• The following day, pipette tips were used to create a straight wound line through the 100% confluent adherent
cell layer. PBS was used to remove the floating cells, and new, fresh RPMI media was added in their stead.

• Using an inverted microscope with a camera, photos of the wound's healing were captured at time points of 0,
24, and 72 hours after the addition of methanolic S. palmetto crude extract to the wells (Nikon, Japan).
4. Cell migration assay ( wound-healing
technique).
Methodology
5. DNA fragmentation.
• The s.palmetto stem extracts were given to MCF7 cancer cells for 6 and 24 hours.

• After incubation, cells were collected using 1X trypsin and centrifuged before being rinsed with PBS buffer.
After that, DNA was isolated from cell pellets using a Promega Genomic DNA purification kit in accordance
with the manufacturer's instructions after being resuspended in 600 µl of lysis buffer (Promega, USA).

• Then of the DNA solution was transferred to a 1.0% agarose gel, and electrophoresis was performed on
the gel for 2 hours at 80 V using a running buffer of 0.5X Tris-borate-EDTA (TBE) buffer.

• Under a UV light source, DNA in the gel was stained with ethidium bromide.
Methodology
5. DNA fragmentation.
 Results
• Cytotoxicity assay (mTT assay)

• Methanol S.palmetto was proven not to specifically destroy MCF7 breast

cancer cells through selective cytotoxicity

• The cytotoxicity of methanol extract against malignant MCF7 cells was low
to moderate (---).
  Cell migration results (angiogenesis)
Results 0 Hours 24 hours 72 hours

Control

S.palmetto
 Results
 DNA fragmentation results (apoptosis)

figure 1: Agarose gel electrophoresis (1.0%) of DNA

extracted from MCF7 cells treated with IC50 of non-
hemolytic methanol extracts of Sabal palmetto plant after
6 h and 24 h incubation. Marker; 1Kb molecular weight
marker. Control; untreated MCF7 cells (control) after 6 h
and 24 h.
 Discussion and Recommendations

1. It would be interesting to analyze the extract at various doses to corroborate the

findings given that the results reveal low cytotoxicity and no anti-cancer impact.

2. Investigating the mechanism underlying the observed angiogenesis effect as well as

the potential Sabal Palmetto Methanolic Extract mechanism of action might be beneficial
in this situation..

3. It would be intriguing to research the extract on different cancer cell lines and compare
the outcomes given the observed activation of apoptosis.
 Discussion and Recommendations
4. Additionally, it would be beneficial to examine the extract's effects on healthy cell lines or
normal cells to determine if they differ from those on cancer cells.

5. Additionally, it would be beneficial to examine the extract on animals because this will provide
insight into the potential benefits and drawbacks of the extract.

6. The extract should also be studied in conjunction with other medications to determine
whether it increases or decreases the potency of those medications.

 Overall, the findings of your study are intriguing, but more research into the effects of
S.palmetto Methanolic Extract on other cell lines and the mechanism of action of the extract
on cancer cells would be helpful.
 Conclusion

• This is the first study to show that S. palmetto stem extracts had anti-cancer properties.

• Extracts from fruits and stems are thought to be a viable source of anti-cancers.

• The methanol extract produced the least cytotoxicity against MCF7 cells and had an IC50
value more than 300, therefore it cannot be regarded as an effective treatment for breast
cancer cells, but it has a remarkable angiogenetic effect on these cells where it prevents
metastasis of the breast cancer cells.

• As a result, stems of S. palmetto are a promising source of physiologically active metabolites

that could be exploited to create new cancer treatments or enhance existing ones.
Biology and biotech-
nology
Thank you