Anticancer effect of lemongrass oil and citral emulsion on cervical cancer cell lines (HeLa and ME-180)

1. Anticancer effect of lemongrass oil and citral emulsion on
cervical cancer cell lines (HeLa and ME-180)
Kavisa Ghosh
Department of Zoology, Annamalai University, Annamalainagar-608 002

2. CANCER
 Heterogeneous group of diseases
 Characterized by:
 Second leading cause of death
 Gender specific

3. CARCINOGENESIS

4. CERVICAL CANCER
• World wide: third most common cancer in women (GLOBOCAN 2008, IARC)
• In India: first threat followed by breast and ovary cancer
(GLOBOCAN 2008, IARC)

5. CERVICAL CANCER
• Malignant tumour in the tissues of the cervix
• Two main types of cervical cancer:
 Squamous cell carcinoma (80%)
 Adenocarcinoma
• Metastasise to other parts of the body through the lymphatic system
• Early changes rarely cause any symptoms
• Doctors encourage women to have regular Pap tests
• Human papillomavirus (HPV) infection is the cause of cervical cancer
• HPV types 16 and 18 are responsible for 70%
• Survival rate:
 Pre-invasive cervical cancer lesions is almost 100%
 Localized cervical cancer- five year survival rate is 92%

6. The Prognosis of Cervical Cancer
HPV Infection Mild cervical
dysplasia
Severe
dysplasis
Cervical
cancer
Extremely
common
among women
of reproductive
age.
A small
percentage of
cases lead to
abnormal cell
changes.
Usually temporary.
Some cases,
however, progress
to serve dysplasia.
Severe
dysplasia is far
less common
than mild
dysplasia. It
can progress to
cancer in 10-15
years.
Most
common
among
women in
their 50s and
60s.

7. East-Indian Lemongrass (Oil density-0.89 g/ml)
Family: Poaceae
Genus: Cymbopogon
Species: flexuosu
Pharmalogical effects:
• Stress-related disorders
• Antifungal
• Antimicrobial
• Anticancer
Bioactive compounds:
• Geranial
• Geranyl acetate
• Neral
• α-bisabolol
• Sesquiterpene isointermedeol

8. CITRAL
• Important constituent of lemongrass oil (75-85 %)
• Other source of citral : Backhousia citrata, Leptospermum citratum,
Ocimum gratissimum , etc.
• Mixture of two isomeric acyclic monoterpene aldehydes
• The trans-citral is known as geranial or citral A
• The cis-citral is known as neral or citral B
• Used in perfumes, flavouring agent and for fortifying lemon oil
• Synthesis of vitamin A, ionone, and methylionone
• Mask the smell of smoke
• Density - 0.89 g/ml
Pharmacological effects:
• Antimicrobial (bactericidal)
• Anticancer
• Insecticidal
• Antifungal

9. AIM OF THE STUDY
The present study aims to evaluate the anticancer effect of lemongrass oil and
citral emulsion on cervical cancer cell lines (HeLa and ME-180) in vitro.
The following aspects were analysed to test this hypothesis:

10. EXPERIMENTAL DESIGN
Group 1:
Control (HeLa)
Group 1:
Control (ME-180)
Group 2:
HeLa + Lemongrass oil emulsion
(200μg/ml)
Group 2 :
ME-180 +Lemongrass oil emulsion
(200 μg/ml)
Group 3 :
HeLa + Citral emulsion (500μg/ml)
Group 3 :
ME-180 + Citral emulsion(300μg/ml)

11. Methods Techniques used
Emulsion preparation Sonicated in MEM for 20 sec
Emulsion size Dynamic light scattering (DLS)
instrument
Cytotoxic effect MTT Assay (Tetrazolium dye)
Intracellular ROS generation 2’,7’- dichlorofluorescin diacetate
(DCFH-DA) dye
Mitochondrial Transmembrane
Potential (ΔΨm)
Rhodamine-123 staining
Apoptotic morphological
changes
Dual staining

13. DETERMINATION OF CITRAL CONTENT
Test Results Obtained Limit As Per IS
327/1991
Citral Content in
lemongrass oil (Sodiun
Bisulphite method)
78.50% 75.0% Minimum
ANALYSIS OF PARTICLE SIZE AND SIZE DISTRIBUTION
Fig. A: Average size of the lemongrass oil emulsion was 267 nm and
Fig. B: average size of the citral emulsion was found to be 270 nm

14. Microscopic images of HeLa cells treated with emulsions for 24 h.
(A)
(B)
(C)
(D)

15. Microscopic images of ME-180 cells treated with emulsions for 24 h.
(A)
(B)
(C)
(D)

16. DISCUSSION
Lemongrass oil and citral emulsion (24 h incubation):

17. SUMMARY

18. CONCLUSION
Lemongrass oil emulsion (200 μg/ml)- inhibit the cell growth in both the
cell lines.
 Citral - 500 μg/ml for HeLa & 300 μg/ml for ME-180.
 Lemongrass oil had higher activity than citral for both the cell lines.
 There may be synergy between the citral and other components with
anticancer activity in lemongrass oil.
Future investigations:
 In vitro anticancer efficacy in in vivo models.
 Understanding the various properties of lemongrass oil and citral emulsions.
 Ointment based anticancer drugs, which can be used in situ.

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