Recurrent somatic DICER1 mutations were found in nonepithelial ovarian cancers. These mutations occurred in codons encoding metal binding sites within the RNase IIIb catalytic centers of DICER1. The mutations were restricted to these sites and did not completely abolish DICER1 function, but instead disrupted microRNA processing in a cell-type specific manner, leading to oncogenesis in certain cell types like ovarian epithelial cells. Validation studies found these DICER1 mutations in a high percentage of additional nonepithelial ovarian tumor samples.

1. RECURRENT SOMATIC DICER1
MUTATIONS
IN NONEPITHELIAL OVARIAN
CANCERS
ALIREZA 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 biology
Universidad Pontificia Bolivariana
2012

2. INTRODUCTION
Cancer is the uncontrolled
growth of abnormal cells in the
body. Cancerous cells are also
Cells are the building
called malignant cells
blocks of living things.
Cancer grows out of Cancer can develop in
normal cells in the almost any organ or
body tissue
Cancer appears to
occur when the growth It can also occur when
of cells in the body is cells forget how to die.
out of control and cells Cell doesn't apoptosis
divide too quickly.

3. INTRODUCTION
There are many
causes of cancers

4. 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.

5. 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.

6. 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.

7. 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.

8. INTRODUCTION

9. 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
http://www.bbc.co.uk/mundo/ciencia_ prevalencia mutaciones DICER1
tecnologia/2009/08/090821_adn_fals
ificaco_pea.shtml

10. MÉTODOS- SECUENCIACIÓN
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
http://proy-genomahumano.blogspot.com/2010/05/proyecto-genoma-
humano.html

11. MÉTODOS- SECUENCIACIÓN
El transcriptoma  conjunto
de ARN mensajero resultante
de la traducción del genoma
El exoma  genoma formado
por los exones  codificantes Los datos fueron depositados
de los genes ARNm  al ser en el archivo Europeo del
traducido dara lugar a una Genoma-Fenoma
proteina
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/

12. MÉTODOS- SECUENCIACIÓN
- Nucleótidos
4.Nucleótidos didesoxi modificados que han
Secuenciación perdido el grupo
(ddATP, ddTTP, ddCTP
Sanger y ddGTP) hidroxilo 3' de la
desoxirribosa
análisis más detallado 5. Nucleótidos 
de la estructura del 3. Cebador o incorporarse a la
ADN averiguar la "primer“, suele ser cadena de ADN
secuencia de marcado naciente, no se une
nucleótidos. por el extremo 3¨
2. Un enzima que 6. Una vez incorporado
1. El ADN molde o
replique el ADN  un nucleótido didesoxi
segmento de ADN que
ADN Polimerasa I del se termina la síntesis
se desea secuenciar
bacteriofago T4. de la cadena de ADN.

13. MÉTODOS - MARCAJE
MARCAJE CON ISOTOPOS:
• Incubacion de la proteina DICER1
• 5'-32phosphate (32P) es un isótopo
1 radioactivo de fósforo
• Etiquetaron sustrato de ARN de oligonucleotidos
• Oligonucleótido secuencia corta
2 de ADN o ARN, 50 PB pares de bases o menos.
• se analizaron los productos de escisión por medio
de electroforesis en gel y fosforimaginacion.
3

14. MÉTODOS - MARCAJE
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 y
este se coloca en una cámara de
electroforesis, que luego se conecta a
una fuente de alimentación
http://www.taringa.net/posts/ciencia-educacion/9792687/Aprende-de-
Electroforesis-y-hacela-vos-mismo_.html

15. MÉTODOS - MARCAJE
Cuando la corriente eléctrica se aplica, las
moléculas más grandes se mueven lentamente
a través del gel mientras que las moléculas
más pequeñas se mueven más rápido
http://javeriana.edu.co/Facultades/Ciencias/neurobioquimica/libros/celular/electroforesis.html

16. MÉTODOS - MARCAJE
PHOSPHORIMAGING-
FOSFORIMAGINACION:
Autorradiografía estándar que
es mucho más precisa en la
cuantificación de la cantidad de
radiactividad en una sustancia.
1. http://quimicoclinico.wordpress.com/2008/02/03/toma-de-muestras-
sanguinea-venosa-y-gases-arteriales/
2. http://escritoriodocentes.educ.ar/datos/catalizadores.html

17. RESULTADOS.

18. RESULTADOS.

19. DISCUSSION
AUTHORS PUBLICATION AGREEMENT
OR DISAGREEMENT
A survey of cancer
cell lines showed that 4 of
Slade I, et al AGREEMENT
781 had truncating
changes in DICER1. 34
Somatic mutations
in DICER1 (albeit not the
Berger MF, et hot-spot mutations
AGREEMENT
al. described here) have been
reported in a single patient
with prostate cancer. 35

20. DISCUSSION
AUTHORS PUBLICATION AGREEMENT
OR DISAGREEMENT
Among 823 patients with a AGREEMENT
variety of primitive tumors,
there was a low prevalence
Slade I, et al
of germline DICER1
mutations that are likely to
cause loss of function.34
Recent studies suggest
that miRNA* species may
Yang JS, Guo be important in gene
AGREEMENT
L, et al regulation rather than
simply being an inert
strand.38,39

21. MAPA CONCEPTUAL
Mariana Tamayo Correa

22. MAPA CONCEPTUAL
María Alejandra Tobón

23. 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/

24. 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/

25. 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.

26. 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