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Molecular Biology Seminary
Evert Jiménez Medicine
24/08/2010
http://formacionbiblioteca.udea.edu.co/moodle/course/view.php?id=143&topic=1
Expression of bone morphogenetic protein-2 and
its receptors in epithelial ovarian cancer and their
influence on the prognosis of ovarian cancer
Patients.
AUTHOR: YING MA
http://formacionbiblioteca.udea.edu.co/moodle/course/view.php?id=143&topic=1
Ma et al. Journal of Experimental & Clinical Cancer Research 2010, 29:85http://www.jeccr.com/content/29/1/85
INTRODUCTION
This study seek determinate the expression of bone morphogenetic protein-2 (BMP-2) and its receptors BMPRIA,BMPRIB, and BMPRII in epithelial ovarian cancer (EOC) and to analyze their influence on the prognosis of ovarian cancerpatients.
CANCERCancer is a mutation or abnormal activation of genes that control cell growth and cell mitosis which end in excess of malignant cells with growth and division beyond the normal limits.
http://www.medicosdeelsalvador.com/Detailed/Im_genes_M_dicas/Coloproctolog_a/C_ncer_de_Ovario_946.html
OVARIOIs the Gonad femenine. is responsible for produce female sex hormones and ovulation. There are two with almond form. They connect with the uterus through the fallopian tubes and suspended by the broad ligament of ovary .
http://www.ferato.com/wiki/index.php/Ovario
BMP-2BMP is a member of the transforminggrowth factor-β superfamily. Initially, it was thoughtto induce bone formation and condrogenesis in vivo,and current evidence suggests that it also participates insome biological processes of cells, such as proliferation,Differentiation and apoptosis.
http://www.treslacteospordia.org.uy/alternativas_chicos.php
The propose of this study is determinate theexpression of BMP-2 and its receptors in epithelial ovariancancer, benign ovarian tumors, and normal ovariantissue and to analyze their influence on the five-year survivalrate and average survival time of ovarian cancerpatients.
GENERAL OBJETIVE
MATERIALES Y METODOS
Amplificación del RNA a través de la síntesis previa de su cDNA, utilizando una transcriptas inversa, seguido de varios ciclos de PCR convencional. Con esta técnica se puede determinar la expresión de genes en diversos tejidos.
Proteínas son separadas por electroforesis en gel según su tamaño. Luego las proteínas se transfieren a un filtro que se incuba con anticuerpos los cuales reaccionan con la proteína de interés y luego se es identificado.
Es la identificación de un tejido por medio de una interacciones especifica entre un antígeno y un anticuerpo ( visiblemente marcado) las células se colorean para demostrar , identificar y localizar la molécula en interés.
RT-PCR
INMUNOHISTOQUIMICA
WESTERN BLOT
MATERIALES Y METODOS
RT-PCR
WESTERN BLOT
INMUNOHISTOQUIMICA
RNA total extraido de 100mg especimenes
1.5% agarosa gel electroforesis conBromuro de etidio.
la proteína fue extraída de 100 mgespecímenesfueron analizadas con el método de BradfordPoliacrilamida gel electroforesisFiltro de nitrocelulosa
Anticuerpos
Parafina
RESULTADO
RT-PCR
WESTERN BLOT
INMUNOHISTOQUIMICA
Positivamente manchada BMP-2 y sus receptores BMPRIA,BMPRIB y BMPRII. Se localizaron principalmente en el citoplasmade las células de cáncer de ovario y apareció como de color marrón claro.
Los niveles de expresión del ARNm de BMP-2, BMPRIB, y BMPRII en los tejidos de cáncer de ovario fue significativamente menor que los de los tumores benignos ováricos o normal tejido. No hubo diferencias significativas en BMPRIA ARNm niveles de expresión que se observaron entre los tres tipos de Tejidos.
El contenido relativo de las proteínas BMP-2, BMPRIB, yBMPRII en el tejido de cáncer de ovario fue significativamente menorque los de los tumores benignos ováricos o normaltejido. No hubo diferencias significativas en proteínas BMPRIAnivel de expresión se observaron entre los tres tipos deTejidos.
Figure 1 The mRNA expression of BMP-2 and its receptors detected by RT-PCR 1: Ovarian cancer tissue; 2: Benign ovarian tumor tissue; 3: Normal ovarian tissue; M: Marker.
1 32
BMP-2
BMRIA
B-actin
BMRII
BMRIB
43KD
75KD
50KD
20KD
66KD
2 The protein expression of BMP-2 and its receptors detected by western blot 1: Ovarian cancer tissue; 2: Benign ovarian tumor tissue; 3: Normal ovarian tissue.
DISCUSSION
http://churchwhisperer.com/2009/09/01/after-much-discussion/
Author What he/she said According with the investigation
Urist MR In 1965, Urist successfully induced heterotopic bone formationby grafting decalcified bovine bone into musclesand skin.
Le Page, et all. detected the expression of BMP-2 in ovarian cancer tissues.
Kiyozuka Y,et all. confirmed that BMP-2 wasinvolved in the formation of serous ovarian cancer psammomabodies.
Miyazono K, et all. BMP cannot act without its receptors, BMPRI (BMPRIA and BMPRIB) and BMPRII, which arelocated on chromosomes 10q23, 4q22-24, and 2q33-34.BMPRIA mediates growth stimulation signals, andBMPRIB transfers growth inhibition signals
CONCLUSION
There is not clarity about if the BMP-2 stimulate the growth of cancerigenas
cell or acts like a protector.
BMP-2 is expressed in ECO, normalovarian tissue and benign ovarian tumor
patients.
Is possible that if BMP-2 is lower or alterated in normal ovario tissue may be
appear ovarian cancer
Is necessary a study with a much larger sample in order to gain more accurate
results
THANK`S
http://www.valledellili.org/sitio/index.php?option=com_content&view=article&id=446%3Aen-el-aparato-reproductor-tambien-hay-riesgo&catid=38%3Anuestras-noticias&Itemid=112&lang=es