seminario biologia
TRANSCRIPT
RECURRENT SOMATIC DICER1
MUTATIONS
IN NONEPITHELIAL OVARIAN
CANCERSALIREZA HERAVI-MOUSSAVI, PH.D., MICHAEL S. ANGLESIO, PH.D.,
S.-W. GRACE CHENG, PH.D., JANINE SENZ, B.SC., WINNIE YANG, B.SC., ET AL
Mariana Tamayo Correa ID: 000192750
María Alejandra Tobón Arango. ID: 000192513
Third Semester – Molecular biologyUniversidad Pontificia Bolivariana
2012
INTRODUCTION
Cancer is the uncontrolledgrowth of abnormal cells in thebody. Cancerous cells are alsocalled malignant cells
Cells are the building blocks of living things.
Cancer grows out of normal cells in the
body
Cancer appears to occur when the growth of cells in the body is
out of control and cells divide too quickly.
It can also occur when cells forget how to die.
Cell doesn't apoptosis
Cancer can develop in almost any organ or
tissue
INTRODUCTION
There are manycauses of cancers
INTRODUCTION
-Malignant tumor that grows in any part of
the ovary. Often in the epithelium.
-Generally, no symptoms.
-Sixth most common malignancy in women.
-High mortality.
-It is more common in women over 50 years.
-It is diagnosed in advanced stages.
-More common in nulliparous women.
INTRODUCTION
DICER1 gene encodes a protein synthesis that
bears his name, DICER1. It is present in many
organisms as Dicer.
Dicer: Is an endoribonuclease of the RNase III
family that cleaves double-stranded RNA
(dsRNA) and pre-microRNA (miRNA) into short
double-stranded RNA fragments.
INTRODUCTION
DICER1 mutations are common in non-
epithelial ovarian cancer, these mutations do not
affect completely the activity of DICER1, except, the
alteration in a kind of specific cells which is a novel
mechanism that disrupts
the microRNA processing that will cause cancer.
These mutations were restricted to the
codons encoding the metal binding sites within
the RNase IIIb catalytic centers, the enzyme
complex from which the DICER1 takes part.
GENERAL OBJECTIVE.
Demonstrate experimentally that DICER1 mutations in
the RNase family IIIb, are largely related to non-
epithelial ovarian tumors predominantly
in Sertolli cells and Leydig, that such mutations are
attributed to the addition of metal fragments in the
catalytic center of RNase IIIb, and that such changes do
not affect completely the function of DICER1, the
impacts are oncogenic only in certain types of cells like
ovarian epithelial cells.
INTRODUCTION
MÉTODOS- MUESTRA
Cohorte: n=14
-Tumores de células de la granulosa juveniles (JGCT)
-Tumores en las células de sertoli y leyding (SLCT)
-Tumores de células primitivas del saco vitelino (PGCTYS)
Validación de la cohorte n=411
Tumores adicionales fueron usados para la validación y la confirmación de la prevalencia mutaciones DICER1http://www.bbc.co.uk/mundo/ciencia_
tecnologia/2009/08/090821_adn_fals
ificaco_pea.shtml
Secuencia:
1. La secuenciación y el análisis de transcriptoma
2. Ejecutaron la captura del exoma de regiones del código genético
3.Datos fueron depositados en el archivo EGAS00001000135
4. Se realizó Secuenciación de Sanger en la región de DICER1
5. DICER1 codifica el dominio RNasa IIIb
MÉTODOS- SECUENCIACIÓN
http://proy-genomahumano.blogspot.com/2010/05/proyecto-genoma-
humano.html
El transcriptoma conjunto de ARN mensajero resultante de la traducción del genoma
El exoma genoma formado por los exones codificantes de los genes ARNm al ser
traducido dara lugar a una proteina
Los datos fueron depositados en el archivo Europeo del
Genoma-Fenoma
http://noticias.lainformacion.com/ciencia-y-tecnologia/ciencias-general/definen-el-transcriptoma-completo-de-la-cianobacteria-
anabaena-decisiva-en-la-fijacion-del-nitrogeno-atmosferico_dhZQytF9OU5WqyElI9eIM3/
MÉTODOS- SECUENCIACIÓN
Secuenciación Sanger
análisis más detallado de la estructura del ADN averiguar la
secuencia de nucleótidos.
1. El ADN molde o segmento de ADN que se desea secuenciar
2. Un enzima que replique el ADN
ADN Polimerasa I del bacteriofago T4.
3. Cebador o "primer“, suele ser
marcado
4.Nucleótidos didesoxi
(ddATP, ddTTP, ddCTPy ddGTP)
- Nucleótidos modificados que han
perdido el grupo hidroxilo 3' de la
desoxirribosa
5. Nucleótidos incorporarse a la cadena de ADN
naciente, no se une por el extremo 3¨
6. Una vez incorporado un nucleótido didesoxise termina la síntesis de la cadena de ADN.
MÉTODOS- SECUENCIACIÓN
MÉTODOS - MARCAJE
1
• Incubacion de la proteina DICER1
• 5'-32phosphate (32P) es un isótopo radioactivo de fósforo
2
• Etiquetaron sustrato de ARN de oligonucleotidos
• Oligonucleótidosecuencia corta de ADN o ARN, 50 PB pares de bases o menos.
3• se analizaron los productos de escisión por medio
de electroforesis en gel y fosforimaginacion.
MARCAJE CON ISOTOPOS:
Técnica de electroforesis en gel: separación de moléculas basadas en el tamaño y la carga, que pueden hacerse pasar a través de un gel hecho de agar o de poliacrilamida.
Las moléculas se dispersan en el gel yeste se coloca en una cámara deelectroforesis, que luego se conecta auna fuente de alimentación
http://www.taringa.net/posts/ciencia-educacion/9792687/Aprende-de-
Electroforesis-y-hacela-vos-mismo_.html
MÉTODOS - MARCAJE
http://javeriana.edu.co/Facultades/Ciencias/neurobioquimica/libros/celular/electroforesis.html
Cuando la corriente eléctrica se aplica, lasmoléculas más grandes se mueven lentamentea través del gel mientras que las moléculasmás pequeñas se mueven más rápido
MÉTODOS - MARCAJE
1. http://quimicoclinico.wordpress.com/2008/02/03/toma-de-muestras-
sanguinea-venosa-y-gases-arteriales/
2. http://escritoriodocentes.educ.ar/datos/catalizadores.html
PHOSPHORIMAGING-FOSFORIMAGINACION:
Autorradiografía estándar que es mucho más precisa en la
cuantificación de la cantidad de radiactividad en una sustancia.
MÉTODOS - MARCAJE
RESULTADOS.
RESULTADOS.
AUTHORS PUBLICATION AGREEMENT
OR DISAGREEMENT
Slade I, et al
A survey of cancer
cell lines showed that 4 of
781 had truncating
changes in DICER1. 34
AGREEMENT
Berger MF, et
al.
Somatic mutations
in DICER1 (albeit not the
hot-spot mutations
described here) have been
reported in a single patient
with prostate cancer. 35
AGREEMENT
DISCUSSION
AUTHORS PUBLICATION AGREEMENT
OR DISAGREEMENT
Slade I, et al
Among 823 patients with a
variety of primitive tumors,
there was a low prevalence
of germline DICER1
mutations that are likely to
cause loss of function.34
AGREEMENT
Yang JS, Guo
L, et al
Recent studies suggest
that miRNA* species may
be important in gene
regulation rather than
simply being an inert
strand.38,39
AGREEMENT
DISCUSSION
MAPA CONCEPTUAL
Mariana Tamayo Correa
MAPA CONCEPTUAL
María Alejandra Tobón
CONCLUSIONS
The identification
of specific sequences of
gene mutations
associated with cancer is
important for epidemiological
studies to determine the early
detection of mutation in order
to find appropriate treatment.
http://www.bubbleinfo.com/2011/10/19/hot-new-idea/
CONCLUSIONS
Mutations in DICER1(Hot
spot)in Nonepithelial Ovarian
Cancers are highly frecuent
(60%) these mutations are in
primitive tumors and
embryonal tumors
suggests involvement of deve
loping cells, these
mutations are rare incancer
patients.
http://www.bubbleinfo.com/2011/10/19/hot-new-idea/
CONCLUSIONS
-People who have mutations
in the germline of DICER1 are predisposed to non-
epithelial ovarian tumors, whose appearance is explained by the
addition of metal fragments to the catalytic center of the
RNase IIIb enzyme by specific sites called "hot-spots" by
inhibiting the expression of miRNA generating a oncogenic
response.
-Non-epithelial ovarian tumors of highest rate
of presentation related to the mutation of DICER1 of
the RNase IIIb family are tumors of Setolli and Lyding
cells.
BIBLIOGRAFÍA
http://by168w.bay168.mail.live.com/default.aspx#!/mail/InboxLight.as
px?n=1723511564!n=731457103&fid=1&fav=1&mid=59a66730-
591e-11e1-94f2-002264c154b4&fv=1
-http://es.wikipedia.org/wiki/Northern_blot
-http://www.cancer.org/Espanol/cancer/ovario/Guiadetallada/
-http://www.ovationsforthecure.org/indexAwareness.php
-http://www.ambrygen.com/tests/dicer1-syndrome