Presentado por: Juana Hoyos Palacio Jennifer Posada Caro
Presentado por: Juana Hoyos Palacio Jennifer Posada Caro
INTRODUCTION
CSN1-CSN8COP9 CSN5
Catalytic center of COP9
Regulation of tumor suppressors ad oncogen
productosstability of intracellular regulators of cell process
Overexpressed in a variety of human cancers
Ubiquitin proteasome pathway
Removal of NEDD8 from CRL
CANCER
Cancer is a potentially fatal disease caused by environmental factors that mutate genes encoding critical cell-regulatory proteins. The resultant aberrant cell behavior leads to an abnormal cell proliferation that destroy surrounding normal tissue and can spread to vital organs resulting in a disseminated disease.
GASTRIC CANCER Gastric cancer (GC ) is an uncontrolled growth of cells in the inner surface of the stomach. Early gastric cancer has no associated symptoms; so most symptoms of gastric cancer reflect advanced disease and by the time they develop, the disease is almost invariably too far advanced for curative procedures.
• Indigestion• Nausea or vomiting• Dysphagia• Postprandial fullness• Loss of appetite• Melena or pallor from anemia• Hematemesis• Weight loss
GASTRIC CANCER Gastrointestinal stromal tumor Carcinoid tumor
Adenocarcinoma
p53P53 is a nuclear phosphoprotein of 53 kDa, encoded by a 2 gene located on the small arm of chromosome 17. It contains five functional domains: N‐terminal domain, Proline rich domain, the large, central core domain, the oligomerization domain and the basic C‐terminal domain.
CSN5
CSN5
Proteasome
APOPTOSIS
OBJECTIVE
Investigate the molecular mechanisms by which CSN5 is involved in gastric cancer.
Materiales y Métodos
Características de los pacientes
Cáncer gástrico +
diagnostico preoperatorio
+ ausencia de tratamiento
preoperatorio con radio o quimioterapia.
Materiales y Métodos Cultivo
Celulas GES + AGS + MKN-45
Cultivo
Anexos 10% suero bobino
37º 5% CO2
Todas las celulas tuvieron un 50% de
crecimiento antes de la transfección
Cultivo
Materiales y Métodos Transfección
CSN5 siRNA siLentFect Lipid Reagent
Non-specific siRNA = control
-se encubaron muestras en un nuevo medio
- después se lisaron
Materiales y Métodos Homogenización
ANGT + GCT Lavado 2 veces con PBS Homogenizacion Centrifugar
Centrifugar sobrenadantes
Remover sobrenadante
Medicion proteinas simples Leupeptina
AprotininaPepsatina
Materiales y Métodos SDS-PAGE Incubación con ac
Western Blot
Materiales y Métodos Variabilidad celular
- 3000 cèlulas de AGS- 2000 cèlulas de MKN45- Ya con su tranfecciòn- Labilidad medida con Cell
count kit
Materiales y Métodos FITC
- Se cultivaron las células del cáncer gástrico con V-FITC
- Se analizò la actividad apoptótica con FACScan Cytometer / microscopio fluorescencte
RESULTADOS
Figura 1
Figura 2
RESULTADOS
Figura 4
RESULTADOS
Figura 4
RESULTADOS
Figura 5
Discusión Author Exposure Opinion
Brady, Gil-Gomez “Bax can be directly regulated by p53 and showed increased expression in certain tissues after apoptotic stimule “
true
Zhang, Chen, Su, Yang, Lee “CSN5 has been reported as a negative regulator of p53”
true
Discusión Author Exposure Opinion
Reed “Bax is a pro-apoptotic member of the Bcl2 famimly proteins, which serve as critical regulators of pathways involved in apoptosis, acting to either inhibit or promote cell death”
True
Roder “Gastric cancer remains one of the leading causes of cancer death worldwide”
True
Conclusion -We think this study is important because now researchers can go more over and think about a key to prevent or fight against gastric cancer
-There are many people that could be beneficiated with this investigation because is another step foreward to know more about gastric cancer, one of the most deathly cancer ever.
-Investigators should apply this investigation to other type of cancer, so they can know more out it and how to handle with it
-Science should invest in new treatments for patient so that they can get more succesfull results in treatment.
Jennifer
Juana