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

INTRODUCTION

• 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.

AIM OF WORK

The aim of present work was to evaluate the effect of theinexpensive medicinal plant, sweet fennel, with cisplatin on cancercervix cell line.

MATERIALS AND METHODS

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 %

RESULTS

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

Electron microscopic study

Controluntreated

50µgciplatin

50µgfennel

80µgfennel

Electron microscopic study

50µg fennel+ 50µg cisplatin

DISCUSSION

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

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.

THANK YOU

Professor Wafaa S. Ramadan