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Vimentin significantly promoted gallbladder carcinoma mestastasis. Authors: Ping Dong, Xiao-wei He, Jun Gu, Wen-guang WU, Mao-Ian Li, Jia-hua Yang, Ling Zhang, Qi-chen Ding, Jian-hua Lu, Jia-Sheng Mu, Lei Chen, Song-gang Li, Liang-fu Ding, Jian-wei Wang, and Ying-bin Liu. Exponents: Angélica Rivera Cermeño and Carolina Sarmiento. UPB Medellín. 2012
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Page 1: Seminariobiology

Vimentin significantly promoted gallbladder carcinoma mestastasis.

Authors: Ping Dong, Xiao-wei He, Jun Gu, Wen-guang WU, Mao-Ian Li, Jia-hua Yang, Ling Zhang, Qi-chen Ding, Jian-hua Lu, Jia-Sheng Mu, Lei Chen, Song-gang Li, Liang-fu Ding, Jian-wei Wang, and Ying-bin Liu.

Exponents: Angélica Rivera Cermeño and Carolina Sarmiento.UPB Medellín.

2012

Page 2: Seminariobiology

CANCER: Is a term used to refer to diseases in wich abnormal cells divide with no control and are able to invade other tissues.

INTRODUCTION:

Taken of : http://reddemedicosorthomolecularylongevidad.blogspot.com/2011/03/el-cancer-conoce-porque-se-forma.html

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GALLBLADDER CANCER:1. Cancer that forms in tissues of the gallbladder.2. Begins in the innermost layer of this tissue and spreads through the outer layers as it grows.3. Is rare, most commonly presented on Native Americans.

INTRODUCTION:

Taken of :http://www.healthgiants.com/2009/11/25/gallbladder-problems-is-surgery-mostly-needed/

Page 4: Seminariobiology

INTRODUCTION:

GALLBLADDER CANCER: Symptomps include:

•Jaundice •Pain above the stomach•Fever•Nausea and vomiting•Bloating•Lumps in the abdomen

Taken of : http://www.beatbloating.com/

Page 5: Seminariobiology

INTRODUCTION:

METASTASIS: The spread of cancer cells from the initial or primary site of disease to another part of the body; also : the process by which such spreading occurs.

/

Taken of :http://trialx.com/curetalk/wp-content/blogs.dir/7/files/2011/05/diseases/Metastasis-3.jpg

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INTRODUCTION:

VIMENTIN:Polypeptide that copolymerizes with other subunits to form the intermediate filament cytoskeleton of mesenchymal cells.

/

Taken of :http://www.abcam.com/Vimentin-antibody-Neural-Stem-Cell-Marker-ab45939.html

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INTRODUCTION:

/

VIMENTIN

Contractility

Cell

Migration Stiffness Proliferation Invasion

Type III cytoeskeleton

protein

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RELATION METASTASIS/VIMENTIN:

INTRODUCTION:

Process of migration of abnormal cells during a divission process

IF protein important in migration Vimentin migration.

Vimentin

Metastasic level

Taken of :http://www.patentdocs.org/patent_profile/Taken of :http://www.virtualmedicalcentre.com/anatomy

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Evaluate vimentin as a possible GBC marker and therapeutic target based on the relation between this protein and GBC-SD and GBC-SD/M3 metastasic level.

GENERAL OBJECTIVE:

/

Taken of :http://www.medcitynews.com/2011/09/virus-for-cancer-treatment-progresses-in-trials-morning-read/b0006421-breast-cancer-cells/

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METÓDOS:

/

1. ¿Porqué?: Son las células que se presentan en alta proporción en el cáncer de vesícula biliar.

2. ¿Para qué? Para hallar la actividad metástasica de las mismas y su expresión de vimentina.

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METÓDOS:

/

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METÓDOS:

/

Tomado de: http://nano.iitm.ac.in/facilities-maldi.htm

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METÓDOS:

Tomado dehttp://www.experimentosnuevos.com/2010/11/

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METÓDOS:

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METÓDOS:

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METÓDOS:

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METÓDOS:

siRNA1, siRNA2, siRNA3

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METÓDOS:

Tomado de: http://medicinew.blogspot.com/2009/11/una-proteina-clave-para-promover-la.html

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RESULTADOS:

• Comparaciónde la potencia

Motilidad

Invasión Célula

Entre

GBC-SD/M3

GBC-SD

Figura 1. Confirmación de la propiedades invasivas de GBC-SD/M3 in vitro.

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RESULTADOS:

Promedio Numero de células invasoras

Campo

Potencia (In vitro)Motilidad

Invasión

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RESULTADOS:Figura 2. Análisis de la metástasis asociada a biomarcadores entre GBC-SDy GBC-SD/M3.

RT-PCR semicuantitativa

Análisis de metástasis

GBC-SD y GBC-SD/M3

Altamente metastasico

Escasamente metastasico

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RESULTADOS:

• Expresión de los genes relacionados con metástasis fue detectada por PCR en tiempo real.

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RESULTADOS:

Figura 3. Proteínas expresadas diferencialmente fueron identificados entre GBC-SD/M3 y GBC SD-utilizando 2-DE y MALDI-TOF-MS.

GBC-SD/M3

GBC-SDComparados

SelecciónPerfiles proteomicos Tres experimentos

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RESULTADOS:

Ampliación de las proteínas expresadas diferencialmente de vimentina y por GBC-SD/M3GBC-SD. C: MALDI-TOF-MS de vimentina.

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RESULTADOS:

Figura 5. Análisis para la expresión vimentina en GBC-SD/M3 yGBC-SD.

RNA total dos células se extrajeron RT-PCR PCR-q Western Blot

Page 26: Seminariobiology

RESULTADOS:

Valores Media Tres experimentos independientes.

* P <0,05 vs GBC-SD.

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RESULTADOS:

Treinta microgramosde células enteraslisadas

10% de SDS-PAGE membrana de PVDF

Detección Nivel de tubulina

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RESULTADOS:

Figura 6. Alteración en la migración y la invasión deVimentina-siRNA GBC-SD/M3 células.

PCR-q Eficacia ARN-Vimentina en GBC-SD/M3

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RESULTADOS:

Western Blot Eficacia Si-ARN-Vimentina GBC-SD/M3

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RESULTADOS:

Migración

Page 31: Seminariobiology

RESULTADOS:

Invasión

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RESULTADOS:

Figura 8. Análisis de Western Blot y estudio inmunohistoquímico de la expresión de vimentina en tumores primarios y metástasis de las muestras de carcinoma de vesícula biliar.

Anti-vimentina anticuerpo monoclonal

+ anticuerpo secundario

Quimioluminiscencia

Page 33: Seminariobiology

RESULTADOS:

MicrofotografíasTumores primarios de carcinoma de vesícula biliar

Metástasis Inmunohistoquímica

Page 34: Seminariobiology

DISCUSSION:

Investigator Statement Student Concept

Omary MB, Coulombe PA, McLean WHI. Pallari HM, Eriksson JE.

“IFs are constructed from two-chain α-helical coiled-coil molecules arranged on an imperfect helical lattice and have been widely used as markers for distinguishing individual cell types within a tissue and identifying the origins of metastatic tumors.”

Agreement

The expression patterns of IFs are cell- and tissue-typespecific, providing each major cell type with a relativelyspecific “fingerprint” of IF proteins.

Page 35: Seminariobiology

DISCUSSION:

Hendrix MJ, Seftor EA, Chu YW.

“However, in some cancers, particularly malignant melanoma and breast carcinoma, there is a strong indication that Vimentin and keratin IFs are coexpressed, thus presenting as a dedifferentiated or interconverted (between epithelial and mesenchymal) phenotype.”

Agreement

The expression of vimentin inepithelial cells is essential to the successive shape change through the interaction with actin, keratin and other IFs.This might be one of the reasons why vimentin is always associated with the migratory status ofepithelial cells.

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DISCUSSION:

Chu YW, Seftor EA, Romer LH, Hendrix MJ. Quan Z, Gu J, Dong P, Lu J, Wu X, Wu W, et al.

“We reported the statistically significant upregulation of vimentin in human metastatic GBC as compared to its paternal low metastatic cell line, and we did some researches about tumor apoptosis mechanism.”

Agreement

Immunoblot and immunohistochemical analysis on 12 human GBC specimens showed consistently increased vimentin expression in metastases compared with primary tumors.

Page 37: Seminariobiology

DISCUSSION:

Hu L, Lau SH, Tzang CH, Wen JM, Wang W, Xie D, et al.

Direct evidence has been provided to link overexpression of vimentin in human hepatocellular carcinoma cells with increased metastasis activity

Agreement

Vimentin screened from comparative proteomic analysis of two GBC cell strains with different metastatic potentials has been shown to be overexpressed by 3.8-fold in high metastatic GBC-SD/M3 cells as compared with low metastatic GBC-SD cells.

Page 38: Seminariobiology

CONCLUSIONES:

• Vimentin tissue level may reflect the metastasis progression in some GBC and may be a useful marker for predicting tumor metastasis.

• Advances in the fields of proteomics give a reason for the discovery of novel markers to improve diagnosis.

• Progress in proteomics studies can be useful to identify therapeutic targets for the treatment of differentes types of cancer.

• These types of studies - like PCR-q and Western Blot- have great clinical importance and are a therapeutic target for the treatment of patients with metastatic GBC.

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MAPS:

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MAPS:

Page 41: Seminariobiology