The document discusses several topics: - Leukemia, its symptoms, and characteristics. It is a cancer of blood forming cells in the bone marrow. - Caspases, which are key proteins that transduce and execute apoptotic signals. They activate initiators and executors of apoptosis. - MAPKs, the three major pathways (ERK, JNK/SAPK, p38MAPK) that regulate cell proliferation, survival, and differentiation. - The objective of the study was to investigate the cytotoxic effects of 15 synthetic flavonoids against human leukemia cell lines and understand the mechanism of cell death, including caspase activation, MAPK pathway activation, and reactive oxygen species generation.

2. INTRODUCTION
LEUKAEMIA
o*Weakness *Fever
•*Tiredness
•*Shortness of breath
•*Infections are more
frequent, more severe
and last longer
•* Purpura, heavy
periods, nosebleeds,
bleeding gums
•*Bleeding and
bruising
Is a cancer which
starts in blood
forming tissue,
usually the bone
marrow. It leads to the
over production of
abnormal white blood
cells. Type of white cell
affected:
-Myeloid or -
Lymphatic
The way the disease
progresses
-Acute or -Chronic
Flavonoids are
natural pigments present
in vegetables, they
protect the body from
damage by oxidizing
agents
PROPIERTIES;
*Antiinflammatory
*Antiviral
*antiallergic effects
*Protective role in heart
diseases, cancer and
different pathologies.

3. INTRODUCTION
CASPASES
Cysteine-aspartic
protease
oAre key proteins for the
transduction and ejecution
of the apoptotic signals
induced by several stimuli.
Are present within the cell
as inactive precursors that
need a proteolytic cleavaje
in order to be active.
Initiators
Executors
Activated by
autoproteolysis when
translocated to specific
cell compartments or
trough the coupling of
adapters and or activators.
Responsible for the final
cleavage of diverse
substrates that mediate
the morphologic changes
during apoptosis.
Among these there are
signalization, DNA
repairing, structure,
transcription proteins,
Mediate intracellular signal transduction in
response to various stimuli and are involved in
the regulation of proliferation, survival and
differentiation
There are three major
MAPK pathways in
mammals:
ERK 1/2
C- Jun NH2-terminal
kinases Stress-activated
protein kinases (JNK/SAPK)
The p38 mitogen-activated
protein kinases (p38MAPK).
MAPKs

4. GENERAL OBJETIVE
• The main aim of this work was to investigate the cytotoxic effects of
15 synthetic flavonoids against human leukaemia cell lines and to
understand the mechanism of cell death of the most active. We
have specifically evaluated the activation of the caspase cascade
and the MAPK pathway as well as the generation of reactive
oxygen species.

5. Cultivo celular y ensayos de
citotoxicidad en líneas
celulares de cáncer humano
• Reducción metabólica del Bromuro de 3-(4,5-
dimetiltiazol-2-ilo)-2,5-difeniltetrazol (MTT) realizada
por la enzima mitocondrial succinato-deshidrogenasa
en un compuesto coloreado de color azul (formazan),
permitiendo determinar la funcionabilidad
mitocondrial de las células tratadas.
• La capacidad de las células para reducir al MTT 
indicador de la integridad de las mitocondrias y su
actividad funcional es interpretada como una medida
de la viabilidad celular.

6. 2 tipos
DIRECTA:
La cantidad de Ac
marcado es
directamente
proporcional a la
cantidad del Ag
presente.
INDIRECTA:
Se hace reaccionar el Ag con la
muestra y después se hace
recaccionar con el Ac marcado.
Ac fluorescente fijado
Ag presente en muestra
Detección de
proteínas por
unión a una
molócula de
fluorocromo

7. Por electroforesis se
separan las
proteínas
Transferidas a una
membrana
absorbente
Se busca la proteína
de interés con
anticuerpos
específicos contra
ella
Unión
antígeno-anticuerpo
Se estudia la
presencia de la
proteína en el
estracto y se analiza
su cantidad con
respecto a otras
proteínas

8. RESULTADOS
Fig. 2. C

9. Fig. 2. D
Fig. 2. E
RESULTADOS

10. RESULTADOS
Fig. 4. AFig. 3. A

11. RESULTADOS
Fig. 5. C
Fig.6. A

12. DISCUSSION
AUTHOR DISCUSSION AGREE DESAGREE
D.S. Bellows, B.N. Chau, P. Lee,
Y. Lazebnik, W.H. Burns, J.M.
Hardwick
“The Bcl-2 cleavage products are potentially proapoptotic
and this event may promote caspase activation and
apoptosis via a mitochondrial pathway.”
D.G. Kirsch, A. Doseff, B.N.
Chau, D.S. Lim, N.C. de Souza-
Pinto, R. Hansford, M.B. Kastan,
Y.A. Lazebnik, J.M. Hardwick,
“The release of cytochrome c by mitochondria is triggered
by the 22- kDa fragment of Bcl-2 generated by caspase-
3.”
M.E. Guicciardi, M. Leist, G.J.
Gores
“Since the broad-spectrum caspase inhibitor did not
totally block the cell death triggered by 11, it is possible
that alternative cell death mechanisms might be involved,
such as the lysosomal pathway which involves a partial
rupture of lysosome membrane and subsequent re- lease
of cathepsins into the cytosol.”

13. CONCLUSIONS
• 1. Compounds or synthetic substances that act on specific points in the
process of apoptosis or on the factors that regulate it, provide starting
points for research in pathologies where there is no control of the
proliferation of cells with uncorrected intracellular damage.
• 2. Because flavonoids can not be synthesized de novo by the human
organism, we would consider that diets rich in these compounds are
favorable for health due to their multiple properties as antiinflammatory
since they exert an inhibition on inflammatory PGs, they also inhibit
thyroid peroxidase. therapeutic action in gout disease, among other
properties and of course its action on the process of apoptosis that take
great importance in cancer diseases.