Haploinsufficiency of Casitas B-Lineage Lymphoma Augments the Progression of Colon Cancer in the Background of Adenomatous Polyposis Coli Inactivation

1. SEMINAR
BY:Santiago Castrillón Montoya
Corina Del Pozo Anderson
UNIVERSIDAD PONTIFICIA BOLIVARIANA
TERCER SEMESTRE
BIOLOGIA MOLECULAR
2020

3. INTRODUCTION
Colorectal cancer (CRC):
Is the third most commonly
diagnosed malignancy in the world.
It still constitutes the fourth leading
cause of cancer-related deaths.
It is a common, lethal, and
prevalent tumor.
CRC-related deaths are
secondary to progression of the
disease and metastases.

4. C-CBL (Casitas B-lineage
lymphoma)
B-Catetin nuclear: signaling
path= WNT
AXIN-2
APC (tumor suppressor
adenomatous polyposis coli)
INTRODUCTION

5. OBJECTIVE
.
Analyze the Haploinsufficiency of Casitas
B-Lineage Lymphoma Augments the
Progression of Colon Cancer in the
Background of Adenomatous Polyposis
Coli Inactivation

6. MÉTODOS

7. PCR
(Reacción de cadena
polimerasa)
- Es una técnica cuyo
objetivo es la amplificación
directa o indirectamente de
un gen o un pedazo de DNA
o RNA.

8. INMUNO
HISTOQUÍMICA
- Tecnica que se basa en una
reaccion entre un antigeno y un
anticuerpo para determinar la
presencia de proteinas celulares.
- Esa reaccion se da sobre un
tejido que es identificada por
medio de un anticuerpo
marcado.

9. With genetic engineering,
we will be able to increase the
complexity of our DNA, and
improve the human race.
Stephen Hawking
RT-PCR of
CBL mRNA
- Se produjo un cDNA
mediante la transcriptasa
inversa
-RNeasy mini kit
- Se midió la concentración
de ese ARNm de cCBL por la
PCR en tiempo real

10. WESTERN
BLOT
-Detecta la cantidad de proteina
expresada al interactuar con un
antigeno especifico
-Luego se miden por una
electroforesis en gel para mirar
la expresión del gen CBL
+/+ HOMOCIGOTO
+/- HETEROCIGOTOGráficas:

11. RESULTADOS

12. Figura 2
A) Los ratones muestran
tumorigénesis espontánea en su
intestino delgado y grueso.
GRÁFICA:
APC 14/+ CBL +/+ homocigotico
APC 14/+ CBL +/- hetorocigotico.

13. Figura 2
C) Los lisados de los testículos
mostraron hasta un 50% de
reducción de la proteína c-Cbl
en los ratones heterocigoticos

14. Figura 2
D) La expresión c-Cbl se redujo
en el epitelio colónico,
especialmente en las células en
los ápices de las criptas

15. Hiperplasia atipica de
intestino delgado
Adenocarcinoma en:
Intestino grueso y
delgado
APC14+/+ c-CBL Homocigoto
APC14-/+ c-CBL Heterocigoto
GRÁFICA:
HAPLOINSUFICIENCIA

16. Analizar el axin2 con
quien interactua la b-
catenina potenciando
su acción en la vía de
señalización Wnt
Flecha amarilla :Citosol
Flecha blanca: núcleo

17. Discussion

18. In combination with
previous data, this work
supports the notion that c-
Cbl down-regulates two
key components of
CRCepithelial
tumorigenesis and tumor-
induced angiogenesis
However, further studies are
needed to elucidate
potential underlying
mechanisms that
compensate for the loss of
ubiquitin ligase, which are
known to undergo
autoubiquitination or
deubiquitination
1.Rahimi N (41)
2. De Bie P,
Ciechanover A
(38)

19. 3. Shasar M (10) In CRC cells, c-Cbl
suppresses the growth of
colon cancer cells by
down-regulating Wnt
signaling and suppressing
nuclear b-catenin

20. CONCLUSION
We conclude that the detection and quantification of
genetic material has shown a great impact in the area of
health, mainly in oncology. The development of tests has
transformed molecular diagnosis into a key tool for the
treatment of future patients.
1.
2. The application of molecular biology has allowed an
important advance in the knowledge on the pathophysiology
of conditions generated by genetic alterations.

21. 3. The ability to generate molecular methods is an interesting
alternative to study, since it allows to know and understand in
a wider way as this possible genetic defect, can become a
specific phenotype.