Chemosensitizing effect of aqueous extract of sweet fennel in cisplatin treated HeLa cells
Wafaa S.Ramadan, Khalid H. Sait, Nisreen M. Anfinan and HeshamSait
Outline
• Introduction• Materials And Methods• Results• Discussion• Conclusion and recommendation• Acknowledgment• References
• Cervical carcinoma
• Clinical and cost effectiveness of cisplatin
• Nephrotoxic effect of cisplatin
The clinical outcomes of cervical cancerpatients treated with cisplatin was based oncisplatin chemo testing in vitro.
• Role of Natural bioactive compounds and herbal remedies
• The value of Sweet fennel (foeniculum vulgaremill.)
• Sweet fennel has exhibited cytotoxicity against five different lymphoblastic cell lines.
The aim of present work was to evaluate the effect of theinexpensive medicinal plant, sweet fennel, with cisplatin on cancercervix cell line.
M&M
Preparation of aqueous extract of
fennel
Culture of HeLa cellsCytotoxicity
test (MTT assay)
Gas chromatography
–mass spectrometry
(GC-MS):
High-performance
liquid chromatograp
hy (HPLC)
Calculation of
combination index
Electron microscopic
studies
Statistical analysis
Experimental design
Ccis Fen80 70 50CisFen80 70 60 5060
The following groups of Hela cells were incubated with cisplatin or fennel for 24hrs.
Group I(C ): untreated Hela cells used as negative control
Group II(cis): treated with cisplatin (50µg /ml).
Group III(Fen): treated with aqueous extract of sweet fennel (50, 60, 70 and 80 µg/ml).
Group IV (CisFen): treated with cisplatin and aqueous extract of fennel simultaneously
Mean O.D of treated cells/Mean O.D of control cells ×100%
Percentage of cell viability
Combination index
𝐶𝐶𝐶𝐶 =cisplatin % + sweet fennel %
cisplatin/sweet fennel %
Table I: Analysis of aqueous extract of fennel using GC-MS and HPLC
Chemical category Composition %
α-pinene 1.5
β-myrcene 0.6
limonene 0.8
Phenyl propanoids 23.0
Hydrocarbons 0.1
PhenolsThe rest are impurties
12.0
Cytotoxicity test
0
20
40
60
80
100
120
Percentage of cell viability 43.6%
85.23%
11.23%
Effect of cisplatin (50/µg/ml) and different concentrations of fennel (50, 60, 70, & 80µg/ml) on the cell viability of HeLa cells. The results are expressed as mean ± SD and were analysed by one –way ANOVA followe by LSD post hoc test.
Table 2: Combination index (CI); analysis of combined treatment of HeLa with sweet fennel and cisplatin for 24 hours.
CI > 1 antagonism; CI < 1 synergism; CI = 1 additive effect
Conc. of cisplatinµg /ml
Conc. of sweet Fennelµg /ml
CI
50 50 0.8260 1.0870 1.1180 1.13
The predominant function of autophagy is to promote cell survival.
Autophagy has been reported to initiate cell death in response to intracellular damage caused by chemotherapeutic agents.
A complex relationship exists between autophagy and apoptosis
CONCLUSIONAND RECOMMENDATION
The combination of cisplatin and 50µg/ml of aqueousextract of fennel could enhance cervical cancer growthinhibition through synergistic action.
This work was supported by grants-in-aid for scientific research from scientific chair of Prof. Abdullah Hussein Basalamah for gynecological cancer at king Abdulaziz University Hospital.
Acknowledgment
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