5- prof.wafaa s.ramadan - presentation for conference -

2. Chemosensitizing effect of aqueous extract of
sweet fennel in cisplatin treated HeLa cells
Wafaa S.Ramadan, Khalid H. Sait, Nisreen M. Anfinan and Hesham
Sait

3. Outline
• Introduction
• Materials And Methods
• Results
• Discussion
• Conclusion and recommendation
• Acknowledgment
• References

4. INTRODUCTION

5. • Cervical carcinoma
• Clinical and cost effectiveness
of cisplatin
• Nephrotoxic effect of cisplatin

6. The clinical outcomes of cervical cancer
patients treated with cisplatin was based on
cisplatin chemo testing in vitro.

7. • Role of Natural bioactive compounds and herbal
remedies
• The value of Sweet fennel (foeniculum vulgare
mill.)
• Sweet fennel has exhibited cytotoxicity against
five different lymphoblastic cell lines.

8. AIM OF WORK

9. The aim of present work was to evaluate the effect of the
inexpensive medicinal plant, sweet fennel, with cisplatin on cancer
cervix cell line.

10. MATERIALS AND
METHODS

11. 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

12. Experimental design
C
cis 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

13. 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 %

14. RESULTS

15. 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
Phenols
The rest are impurties
12.0

16. 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.

17. 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 cisplatin
µg /ml
Conc. of sweet Fennel
µg /ml
CI
50 50 0.82
60 1.08
70 1.11
80 1.13

18. Electron microscopic study
Control
untreated
50µg
ciplatin
50µg
fennel
80µg
fennel

19. Electron microscopic study
50µg fennel+ 50µg
cisplatin

20. DISCUSSION

21.  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

22. CONCLUSION
AND RECOMMENDATION
The combination of cisplatin and 50µg/ml of aqueous
extract of fennel could enhance cervical cancer growth
inhibition through synergistic action.

23. 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

24. BIBLIOGRAPHY
• Dupont, N.C. And B.J. Monk, chemotherapy in the management of cervical carcinoma. Clin adv
hematol oncol, 2006. 4(4): p. 279-86.
• Aggarwal, b.B., Et al., From traditional ayurvedic medicine to modern medicine: identification of
therapeutic targets for suppression of inflammation and cancer. Expert opin ther targets, 2006. 10(1): p.
87-118.
• Pharmacopia, e., <Eu pharmacopia - contents of the 5th edition.Pdf>, in europian eu pharmacopia.
2005.
• Jakubowicz-gil, j., Et al., Cell death in hela cells upon imperatorin and cisplatin treatment. Folia
histochem cytobiol, 2012. 50(3): p. 381-91
• Kondo, y., Et al., The role of autophagy in cancer development and response to therapy. Nat rev cancer,
2005. 5(9): p. 726-34.

25. THANK YOU
Professor Wafaa S. Ramadan