1. The study investigated the effects of casticin on DNA damage and repair in human lung cancer A549 cells. 2. Casticin suppressed the total viable cell number of A549 cells by inducing DNA condensation and damage, leading to cell death and reduced viability. 3. Casticin induced DNA fragmentation in A549 cells, related to reduced levels of DNA damage and repair proteins, through cytotoxic effects and reduced viable cell numbers.

1. P R E S E N T A D O
P O R : D A N I E L A C A R O L I N A V I L L A L B A C E L L I Y
J O R G E E N R I Q U E Y A B U R B E D O Y A
U N I V E R S I D A D P O N T I F I C I A B O L I V A R I A N A
P R E G R A D O M E D I C I N A
T E R C E R S E M E S T R E .

2. F I R S T A I D
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An Introduction
DEFINITIONS
Small cell lung cancer (SCLC) : is the more aggressive
form of lung cancer.The cancer cells tend to grow
quickly and travel to other parts of the body, or
metastasize
Non-small cell lung cancer (NSCLC):It starts when
healthy cells in the lung change and grow out of control,
forming a mass called a "tumor," "lesion," or "nodule."
1.- Lung cancer: type of cancer that begins in the lungs. Is
the single most significant cause of cancer mortality .
worldwide.
2.- A549 cells: this are the cells that cause Human lung
carcinoma.

3. F I R S T A I D
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4. F I R S T A I D
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An Introduction
DEFINITIONS
1.- Casticin : is an active flavonoid , isolated from a chinese
herb called Vitex Fructus , that is used to treat inflamation in
china , its
been
reported in various bilogical and pharmacological activities
2.- DNA damage: alteration in DNA structure that is capable
of causing celullar injury and reduces viability or reprodutive
fitness of
the
organism
3.- BRCA1 and p53: tumor supressor gene and protein.

5. F I R S T A I D
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6. Objetive
Investigate the effects and molecular
mechanism of casticin on DNA damage and
repair in human lung cancer A549 cells
F I R S T A I D
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7. Methods

8. COMET ASSAY
Capable of measuring DNA damage and repair at
individual cell level, it uses the capacity if negative
charged fragments of DNA to be pulled through an
agarose gel in response to an electric field, looking
like a comet.
It was used to evaluate DNA damage in the study.
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9. WESTERN BLOTTING
Involves the use of gel eletrophoresis to separate
proteins from the sample, the separeted proteins
are trasnfered from the gel to the surface of a
membrane, the membrane is exposed to a specific
antibody against the protein under study, the
antibody is detected using a chemical or
radioactive marker.
Used to examine the effects of caticin on the levels
of DNA damage asociated proteins in the study.
F I R S T A I D
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10. INMUNOFLUORESCENCE ASSAY
Involve an immunostaining that makes use of
antibodies chemically bound to a fluorescent
substance to demonstrate the presence of a
certain molecule
It was applied to detect the expression and
translocation of p-H2A.X by focal spectral
microscopy.
F I R S T A I D
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11. DNA GEL ELECTROPHORESIS
It consists on, applying a current through a gel
containing the molecules of interest. Based on
their size and charge, the molecules will travel
through the gel in different directions or at
different speeds, separating them from one
another.
It was applied to detect de DNA damage of cells
A549 cells treated with casticin .
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12. RESULTS

13. RESULTS

15. Conclusion
1.- Casticin suppressed the total viable cell number of A549, with
DNA condensation and damage, leading to cell death, reducing
A549 cell viability
2.- Casticin induced DNA fragmentation in A549 cells in vitro,
related to the protein level of DNA damage and repair in the cells.
Reducing the viable cell numbers.
3.- Induced cytotoxicity and reduced cell proliferation in human
breast cancer, gastric cancer and myeloma. And decreased lung
cancer by inducing apoptosis by activating FoxO3a.

16. Thank you for the
attetion!
Gracias!

17. C A S T I C I N S U P P R E S S E D T H E T O T A L V I A B L E C E L L
N U M B E R O F A 5 4 9 , W I T H D N A C O N D E N S A T I O N A N D
D A M A G E , L E A D I N G T O C E L L D E A T H :
E X E R T S A N T I - I N F L A M M A T O R Y E F F E C T A N D I T
R E D U C E D T H E L E V E L S O F I L - 6 T U M O R N E C R O S I S
F A C T O R . P R O D U C I N G L A T E R C I T O T O X I C E F F E C T S
V I A R E D U C T I O N O F V I A B L E N U M B E R S A N D
I N F ¿ D U C T T I O N O F D N A D A M A G E
I N D U C E D C Y T O T O X I C I T Y A N D R E D U C E D
C E L L P R O L I F E R A T I O N :
B Y B L O C K I N K T H E P H O S P H O R Y L A T I O N O F
A K T A N D M T O R , P R O T E I N S , I N D I C A T I N G
T H E I N V O L V E M E N T O F A K T / M T O R
S I G N A L I N G P A T H W A Y .
A N D P R O M O T E D F O X 3 A
D E S P H O S P H P R Y L A T I O N A N D F O X M 1
I N A C T I V A T I O N , L E D I N G T O G R O W T H
S U P P R E S S I O N A N D C E L L C Y C L E A R R E S T .
C A S T I C I N I N D U C E D D N A F R A G M E N T A T I O N I N A 5 4 9
C E L L S :
C A S T I C I N I N D U C E D T H E C O N C E N T R A T I O N O F D N A
A N D L A T E R P R O D U C E D D A M A G E ( B Y D N A
F R A G M E N T A T I O N . L E A D I N G T H E C E L L T O C Y C L E
A R R E S T A N D L A T E R A P O P T O S I S
W E S T E R N B L O T T I N G
D N A G E L E L E C T R O P H O R E S I S
C O M E T A S S A Y
I N M U N O F L U O R E S C E N C E A S S A Y
I M P O R T A N T M E T H O D S :
1 .
2 .
3 .
4 .
L U N G C A N C E R : T Y P E O F C A N C E R T H A T B E G I N S
I N T H E L U N G S .
D E F I N I T I O N S :
1 .
2 . A 5 4 9 C E L L S : T H I S A R E T H E C E L L S T H A T C A U S E
H U M A N L U N G C A R C I N O M A .
3 . C A S T I C I N : I S A N A C T I V E F L A V O N O I D , T H A T I S
U S E D T O T R E A T I N F L A M A T I O N I N C H I N A
4 . D N A D A M A G E : A L T E R A T I O N I N D N A S T R U C T U R E
T H A T I S C A P A B L E O F C A U S I N G C E L U L L A R I N J U R Y
Daniela Villalba Celli