Oncogenes y Antioncogenes Dr. Daniel R. CIOCCA IMBECU ...
Oncogenes y Antioncogenes Dr. Daniel R. CIOCCA IMBECU ...
Oncogenes y Antioncogenes Dr. Daniel R. CIOCCA IMBECU ...
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<strong>Oncogenes</strong> y <strong>Antioncogenes</strong><br />
<strong>Dr</strong>. <strong>Daniel</strong> R. <strong>CIOCCA</strong><br />
<strong>IMBECU</strong> (Unidad Ejecutora del CONICET)<br />
Fund. Argentina Investigación Cáncer<br />
MENDOZA
Oncogénesis Mamaria<br />
INICIADORES - Alteración genética<br />
Mutación hereditaria (BRCA1, etc..) o Adquirida<br />
(radiaciones, metabolismo endógeno, químicos, fármacos,<br />
virus, hormonas)<br />
oncogenes-antioncogenes<br />
PROMOTORES – Expanden alt. genética<br />
Hormonas<br />
FACILITADORES – Disminuyen defensas<br />
Inmunodepresores (stress, fármacos, enfermedades,<br />
hormonas)
<strong>Oncogenes</strong><br />
Proto-oncogenes: . Multiplicación celular<br />
. Diferenciación<br />
. Muerte/sobrevida<br />
Alteración dominante<br />
. Mutaciones puntuales<br />
. Amplificación (múltiples copias de ADN)<br />
. Translocación (rearreglos estructurales)<br />
<strong>Oncogenes</strong>
Daños endógenos Daños exógenos<br />
Durante la replicación del ADN:<br />
incorporación de U<br />
Desaminación de A, G o C<br />
Hidrólisis de uniones N-glucosílicas:<br />
despurinación/despirimidación<br />
Oxidación de bases: ERO<br />
Radiaciones: ionizante y UV<br />
Carcinógenos químicos:<br />
benzopireno,<br />
hidrocarburos aromáticos policíclicos<br />
<strong>Dr</strong>ogas antineoplásicas
Ph cromosome, CML<br />
t(9;22)(q34;q11)<br />
Abl codifica tirosina quinasa<br />
que cuando se fusiona con<br />
Bcr es constitutivamente activa<br />
Cromosomas involucrados Bandas afectadas por los puntos de ruptura
Genes Supresores Tumorales (<strong>Antioncogenes</strong>):<br />
. Multiplicación celular<br />
. Reparación daños al ADN<br />
. Muerte/sobrevida<br />
Mutaciones recesivas<br />
Metilaciones<br />
Pérdida del Antioncogen<br />
Cascada de Defectos del Genoma/Proteoma
P<br />
HPV-16<br />
E7<br />
Rb-Mediated Regulation of Cyclin E Transcription<br />
Rb<br />
HDAC-1<br />
P<br />
Cyclin<br />
CDK<br />
Pol II<br />
Pol II<br />
HDAC-1<br />
Rb<br />
E2F<br />
E2F<br />
T-Ag<br />
Cyclin E<br />
CDK 2<br />
HDAC-1<br />
Rb<br />
T-Ag
Rayos X<br />
Radicales libres<br />
Agentes alquilantes<br />
Reacciones espontáneas<br />
U G<br />
G<br />
Uracilo<br />
Sitio abásico<br />
8-oxoguanina<br />
Rupturas de simple cadena<br />
T<br />
Luz UV<br />
Hidrocarburos<br />
aromáticos<br />
policíclicos<br />
T<br />
T<br />
(6-4)PP<br />
Aductos<br />
DCP<br />
C<br />
Rayos X<br />
Antineoplásicos<br />
(Cisplatino)<br />
G G<br />
Uniones cruzadas<br />
Rupturas de doble cadena<br />
Errores de<br />
replicación<br />
A<br />
G<br />
C<br />
T<br />
Bases mal apareadas<br />
Inserciones<br />
Deleciones<br />
BER NER HR, EJ MMR<br />
Síndromes con reparación<br />
del ADN defectuosa<br />
HSP in DNA repair<br />
Nadin & Ciocca<br />
In press<br />
Síndrome Mecanismo gen/proteína Inestabilidad Relación<br />
afectado afectada genómica con cáncer<br />
Xeroderma NER (+/- TCR) XPC/XPC Mutaciones de piel<br />
Pigmentosum XPA/XPA puntuales inducido por UV<br />
(XP) XPD/XPD o por exposición a<br />
XPG/XPG agentes químicos<br />
XPF/XPF<br />
HNPCC MMR hMLH1/hMSH2 Mutaciones Cáncer colorectal<br />
hMSH6/hPMS1/hPMS2 puntuales<br />
BRCA1/BRCA2 HR BRCA1/BRCA2 Aberraciones Cáncer de mama<br />
cromosómicas (ovario)<br />
Ataxia Telangiectasia DSB repair ATM Aberraciones Linfomas<br />
cromosómicas
Genomic analysis of a spontaneous model of breast cancer metastasis<br />
to bone revels a role for the extracellular matrix<br />
Eckhardt BL et al (Robin Anderson). Peter MacCallum Cancer Center, Australia
Primary tumour expression of the cysteine cathepsin inhibitor Stefin A inhibits<br />
distant metastasis in breast cancer<br />
BS Parker, DR Ciocca, et al<br />
Journal of Pathology 214:337-346 (2008).<br />
Cysteine cathepsin inhibitor Stefin A as a gene differentially expressed in primary and<br />
metastatic mammary tumours.<br />
In primary tumours, Stefin A expression correlated inversely with metastatic potential<br />
in 4T1-derived lines and was not detected in tumour cells in culture, indicating induction<br />
only within the tumour microenvironment. Enforced expression of Stefin A in the highly<br />
metastatic 4T1.2 cell line significantly reduced spontaneous bone metastasis following<br />
orthotopic injection into the mammary gland.<br />
There have been no reports of Stefin A expression in metastases in vivo and the relative<br />
expression of Stefin A and the cysteine cathepsins, including their cellular localization,<br />
has not been studied in bone metastasis. In this report, we document the expression and<br />
localization of Stefin A in mouse and human breast tumours and provide functional<br />
evidence for its potential role as a prognostic/therapeutic in advanced breast cancer.
RNA isolated from primary tumours was<br />
reverse-transcribed and real-time RT–PCR was<br />
used to detect Stefin A1, A2 and A3 expression<br />
relative to GAPDH. RTA, relative transcript<br />
abundance. ∗∗p < 0.05 (4T1 to 4T1.2/ 4T1.35)<br />
Stefin A reduces spontaneous metastasis to bone. Stefin A1-containing or<br />
BV-retroviral vectors were used to infect 4T1.2neo1 tumour cells.<br />
A.- Real-time RT–PCR and western blot detection of Stefin A1 RNA and protein expression (flag tag<br />
protein) in pooled and single cell clones.<br />
B.- RT–QPCR detection of tumour burden in spine (using neomycin-tagged 4T1.2 cells) in mice<br />
injected with 4T1.2 BV or 4T1.2 StfA1. RTB is the relative tumour burden (left) and plasma<br />
calcium serum concentration (mg/dl) in tumour-bearing and non-tumour-bearing mice (right).
PEFF sections of reduction mammoplasty tissue, primary<br />
breast tumours and metastases in lung and bone were stained<br />
with mouse anti-human Stefin A or 1B5 hybridoma<br />
supernatant control and visualized with DAB.<br />
Kaplan–Meier analysis comparing disease-free<br />
survival (DFS) (B)<br />
and death due to cancer (DDC) (C)
Co-expression of Stefin A and cathepsin B in lung and bone metastases. Stefin A fluorescence is visualized in red and<br />
cathepsin B in green (E), and these are shown both separately and as a merged image. Sections were additionally stained<br />
with PI (coloured blue) to visualize the nuclei (F).<br />
Final model selected using Akaike information criteria. The model was<br />
initially fitted with all variables (Table 1), including Stefin A staining.
CONCLUSIONS<br />
In a multivariate disease-free survival analysis (Cox proportional hazards<br />
model), Stefin A expression remained a significant independent prognostic factor<br />
in patients with invasive ductal carcinoma (p = 0.0014), along with grade and<br />
progesterone receptor (PR) status.<br />
We propose that Stefin A, as a cysteine cathepsin inhibitor, may be a marker of<br />
increased cathepsin activity in metastases.<br />
Using immunohistology, the cathepsin inhibitor was detected co-expressed with<br />
cathepsin B in lung and bone metastases in both the murine model and human<br />
tissues.<br />
We conclude that Stefin A expression reduces distant metastasis in breast cancer<br />
and propose that this may be due to the inhibition of cysteine cathepsins, such as<br />
cathepsin B.
Multi-step mammary tumor progression<br />
C3(1) SV40 Tag Transgenic Model<br />
NL Dysplasia DCIS Inv Ca Met Ca<br />
1 month<br />
p53 + Rb<br />
Inactivation<br />
6 months<br />
(100% Incidence)<br />
? ? ?<br />
Maroulakou et al. Proc. Natl. Acad. Sci. USA 91:11236, 1994
Categorización de los Eventos<br />
Moleculares en Cáncer de Mama<br />
Porteros: BRCA, p53, Rb<br />
Esporádicos: HER-2/neu, c-myc, ras<br />
Otros: Bcl-2, Hsps, ciclinas, VEGF<br />
Daño al ADN,<br />
alteración ciclo celular<br />
Mayor estímulo para<br />
crecimiento<br />
Falla apoptosis, y otras<br />
moléc., angiogénesis
Carga<br />
genética Daños innatos al ADN<br />
Daños adquiridos<br />
Fallas controles de proliferación<br />
Fallas controles de apoptosis<br />
Edad<br />
Cáncer<br />
Declinación de la inmunidad<br />
Cambios hormonales<br />
Disminución mecanismos reparativos<br />
Acumulación de daños al genoma/proteoma