The document evaluates the anticancer activity of Semecarpus anacardium methanolic extract on skin cancer cell lines. It finds that the extract has an IC50 of 200 μg/ml for inhibiting A431 skin squamous cell carcinoma cells, which is a lower dose than the targeted drug imatinib. Morphological analysis and staining shows the extract changes cancer cell morphology and induces apoptosis through reactive oxygen species generation, but does not affect normal skin cells even at higher doses. The results suggest the extract has anti-cancer activity in skin cancer cells while being non-toxic to normal cells.

1. Supervisor
Dr. Snober S. Mir
Associate Professor
Department of Biosciences
Integral University
Lucknow.
Presented by
Saba Sheikh
Ph.D. Scholar
Department of Biosciences
Integral University
Lucknow.
Evaluation of Anticancer activity of Semecarpus anacardium
methanolic extract on Skin cancer cell line

2. INTRODUCTION
 Cancer is one of the major public health problems worldwide and accounts for an
estimated 2.5 million cases in India alone.
The incidence rate of skin cancer entities is dramatically increasing worldwide.
About 2 out of 10 skin cancers are squamous cell carcinoma and is the second most
common skin cancer
Squamous cell carcinoma of skin tend to grow and likely to invade fatty tissues just
beneath the skin, and are more likely to spread to lymph nodes and/or distant parts of the
body.

3.  In the wake of resistance to chemotherapy and escalating side effects of
synthetic drugs/compounds, all possible avenues are being explored to develop
potential anticancer drug for skin cancer.
Much research has been geared towards the evaluation of plant extracts as
prophylactic agents, which offer great potential to inhibit the carcinogenic process.
Extracts may include more active agents, such as vitamins, minerals, alkylating
agents, anti-metabolites and anti-oxidants and which have potential for the
management of several human clinical conditions.
………..

4. OBJECTIVES
The objectives of the study which are as follows is to test the anti-cancer activity of
promising Indian medicinal plant such as Semecarpus anacardium L. on A431 skin SCC
cell line as well as their effects on HaCaT normal skin cell line
To quantitate the anti-proliferative activity of the extracts.
To identify the apoptosis induced by the selected medicinal plant extracts.
To determine the effect of extracts on ROS generation.

5. MATERIAL AND METHODS
 Preparation of Semecarpus anacardium nut methanolic extract by Soxhlet extraction.
Sub-culturing of A431 skin caner cell line and HaCaT cell line procured from NCCS
 MTT assay for quantification of proliferation of cells after treatment
 Morphological analysis by Heamatoxylin/Eosin Staining
 Propidium Iodide staining for apoptosis detection.
DCFDA staining for ROS generation.

6. RESULTS
Graph A show % inhibition of A431 skin SCC line by Semecarpus anacardium L.
methanolic extract at 24hrs.
The IC50 value is 200 µg/ml
 MTT ASSAY

7. RESULTS
 MTT ASSAY In Positive control and in HaCaT normal cell line
Graph C shows the cytotoxicity to cells
after using a molecular targeted drug
Imatinib as positive control IC50 at a
dose of 600µg/ml
C)
D)
Graph D shows that there is no significant
inhibition of normal cell after treatment of
much higher dose of plant extracts

8. RESULTS
Morphological analysis By Heamatoxylin/Eosin staining
A-Control
B- 200µg/ml S. anacardium
C-Control HaCaT
D- 400µg/ml S. anacardium
Morphological analysis showing the anti-proliferative effect of extract at IC50 dose in
comparison to cancerous and normal controls.
Also showing no effects on normal skin cells HaCaT at higher (400µg/ml) doses of extracts
A B
C D

9. The Pictures Showing the PI+ cells in treated cells but no PI+ in Normal as
well as in cancerous control
A-Control A431 cells
B-200µg/ml of S. anacardium
G-Control HaCaT normal cells
H-400µg/ml of S. anacardium
PI staining for apoptosis detection
RESULTS

10. RESULTS
DCFDA staining for ROS
A-Control A431 skin SCC
B-200µg/ml S. anacardium
C- Control HaCaT normal skin
D-400 µg/ml S. anacardium
These picture showing that the fluorescence is more in treated cells as compared to cancer
control which indicate the generation of ROS .
Interestingly florescence is not seen in normal control as well as in treated normal cells
which shows the absence of ROS.
A B
C D

11. CONCLUSION
 It was found by the result of MTT assay that IC50 dose of S. anacardium is 200µg/ml
which is much lesser dose in comparison to a molecular targeted drug imatinib IC50
dose 600µg/ml in A431 skin SCC cell line.
It is also observed by MTT assay of normal treated cells that extract is not cytotoxic
while using their higher dose i.e 400µg/ml.
It is found by the H/E staining that the extracts changes the morphology of cancer
treated cells but exerting no effect on normal cells at higher dose.
DCFDA staining pic. shows, that the extract liberated ROS which may be a cause of
Apoptosis in cancer cells as identified by PI staining but its absence in normal treated
cell showing extracts non-toxicity.
It can be concluded by the results, that the extracts showing the anti-cancer activity by
altering the morphology, inducing apoptosis and liberating ROS in cancer treated cells
while non toxic to the normal cells.

12. ACKNOLEDGEMENT

13. THANK YOU