Name: models of bone metastasis
Uploaded: 2012-09-04T13:00:00
Description: Watch this Scientific Journal Video about Models of Bone Metastasis at JoVE.com

Forfatterne anerkender følgende finansieringskilder: P01CA040035 (FE / JAS) og VA Career Development Award (JAS).

1. Cell Vedligeholdelse <ol><li> Mange knogle metastatiske subkloner af MDA-MB-231 findes der har variable tilbøjelighed til knogle kolonisering og vækst. Til vore eksperimenter anvender vi en klon afledt af GFP-mærkede MDA-MB-231 udviklet på UTHSCA 2. Denne særlige sub-klon vil danne synlige osteolytiske knoglelæsioner på cirka 3 uger efter inokulering med aflivning efter 4 uger. </li><li> Opretholde cellerne i DMEM indeholdende 10% FBS og 0,1 mg / ml penicillin-streptomycin ved 37 ° C i 5-8% CO <s...

<ol>
	<li>Sterling, J. A., Edwards, J. R., Martin, T. J., Mundy, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+the+biology+of+bone+metastasis:+how+the+skeleton+affects+tumor+behavior.">Advances in the biology of bone metastasis: how the skeleton affects tumor behavior.</a> Bone. 48, 6-15 (2011).</li><li>Yoneda, T., Sasaki, A., Mundy, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteolytic+bone+metastasis+in+breast+cancer.">Osteolytic bone metastasis in breast cancer.</a> Breast Cancer Res. Treat. 32, 73-84 (1994).</li><li>Johnson, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TGF-beta+promotion+of+Gli2-induced+expression+of+parathyroid+hormone-related+protein,+an+important+osteolytic+factor+in+bone+metastasis,+is+independent+of+canonical+Hedgehog+signaling.">TGF-beta promotion of Gli2-induced expression of parathyroid hormone-related protein, an important osteolytic factor in bone metastasis, is independent of canonical Hedgehog signaling.</a> Cancer Res. 71, 822-831 (2011).</li><li>Li, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+TGF-beta+Responsiveness+in+Prostate+Stromal+Cells+Alters+Chemokine+Levels+and+Facilitates+the+Development+of+Mixed+Osteoblastic/Osteolytic+Bone+Lesions.">Loss of TGF-beta Responsiveness in Prostate Stromal Cells Alters Chemokine Levels and Facilitates the Development of Mixed Osteoblastic/Osteolytic Bone Lesions.</a> Mol. Cancer Res. , (2012).</li><li>Yoneda, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Actions+of+bisphosphonate+on+bone+metastasis+in+animal+models+of+breast+carcinoma.">Actions of bisphosphonate on bone metastasis in animal models of breast carcinoma.</a> Cancer. 88, 2979-2988 (2000).</li><li>Rose, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteoactivin+promotes+breast+cancer+metastasis+to+bone.">Osteoactivin promotes breast cancer metastasis to bone.</a> Mol. Cancer Res. 5, 1001-1014 (2007).</li><li>Sterling, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hedgehog+signaling+molecule+Gli2+induces+parathyroid+hormone-related+peptide+expression+and+osteolysis+in+metastatic+human+breast+cancer+cells.">The hedgehog signaling molecule Gli2 induces parathyroid hormone-related peptide expression and osteolysis in metastatic human breast cancer cells.</a> Cancer Res. 66, 7548-7553 (2006).</li><li>Lu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VCAM-1+promotes+osteolytic+expansion+of+indolent+bone+micrometastasis+of+breast+cancer+by+engaging+alpha4beta1-positive+osteoclast+progenitors.">VCAM-1 promotes osteolytic expansion of indolent bone micrometastasis of breast cancer by engaging alpha4beta1-positive osteoclast progenitors.</a> Cancer Cell. 20, 701-714 (2011).</li><li>Oyajobi, B. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+myeloma+in+skeleton+of+mice+by+whole-body+optical+fluorescence+imaging.">Detection of myeloma in skeleton of mice by whole-body optical fluorescence imaging.</a> Mol. Cancer Ther. 6, 1701-1708 (2007).</li><li>Johnson, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+live+animal+micro-CT+allows+for+quantitative+analysis+of+tumor-induced+bone+destruction.">Longitudinal live animal micro-CT allows for quantitative analysis of tumor-induced bone destruction.</a> Bone. 48, 141-151 (2011).</li><li>Sterling, J., Johnson, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Imaging Techniques for the Detection and Examination of Breast Cancer Metastasis to Bone. 46, (2011).</li><li>Steinbauer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GFP-transfected+tumor+cells+are+useful+in+examining+early+metastasis+in+vivo,+but+immune+reaction+precludes+long-term+tumor+development+studies+in+immunocompetent+mice.">GFP-transfected tumor cells are useful in examining early metastasis in vivo, but immune reaction precludes long-term tumor development studies in immunocompetent mice.</a> Clin. Exp. Metastasis. 20, 135-141 (2003).</li><li>Oyajobi, B. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual+effects+of+macrophage+inflammatory+protein-1alpha+on+osteolysis+and+tumor+burden+in+the+murine+5TGM1+model+of+myeloma+bone+disease.">Dual effects of macrophage inflammatory protein-1alpha on osteolysis and tumor burden in the murine 5TGM1 model of myeloma bone disease.</a> Blood. 102, 311-319 (2003).</li></ol>

Undersøgelser af tumor-induceret knoglesygdom afhængige af adskillige dyremodeller, hvoraf de to mest almindeligt anvendte er intrakardiale og intratibial vaccinationer. I mange tilfælde intrakardiale er den bedste mulighed for at studere metastaser til knogler, men betyder det ikke repræsentere den fulde metastatisk proces, og derfor yverfedt pad eller andre ortotopisk injektioner bør ideelt set anvendes mod herpå. Tumorceller ikke metastaserer let fra det primære sted i de fleste undersøgelser med humane celle...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td></tr><tr><td> MDA-MB-231 humane brystcancerceller </td><td> Oprindeligt ATCC-Internt klonet og valgt </td><td></td></tr><tr><td> DMEM </td><td> GIBCO </td><td> 11.885 </td></tr><tr><td> PBS </td><td> Mediatech Inc </td><td> 21-040-CV </td></tr><tr><td> FBS </td><td> Atlas Biologisk </td><td> F-0050a </td></tr><tr><td> Trypsin </td><td> GIBCO </td><td> 15.400 </td></tr><tr><td> Faxitron </td><td> Faxitron </td><td></td></tr><tr><td> Maestro </td><td> CRi </td><td></td></tr><tr><td> μCT scanner </td><td> Scanco </td><td></td></tr><tr><td> Athymiske nøgne mus </td><td> Harlan </td><td> Fox nu - / - </td></tr><tr><td> Insulinsprøjter (300 ul, 28,5 g) </td><td> Becton Dickinson </td><td> 309.300 </td></tr><tr><td> IVIS </td><td> Caliper </td><td></td></tr><tr><td> Bestrålede Rodent Diet </td><td> Teklad </td><td> 2920x </td></tr></tbody></table>

models of bone metastasis

Knoglemetastaser er en almindelig forekomst i flere maligniteter, herunder bryst-, prostata og lunge. Når de er etableret i knogler, tumorer er ansvarlige for signifikant morbiditet og mortalitet 1. Der er således et stort behov for at forstå de molekylære mekanismer, der styrer etablering, vækst og aktivitet af tumorer i knogler. Adskillige in vivo modeller er blevet etableret for at studere disse begivenheder, og hver har særlige fordele og begrænsninger. Den mest almindeligt anvendte model anvender intrakardiale inokulering af tumorceller direkte i den arterielle blodtilførsel af athymiske (nøgne) mus BalbC. Denne procedure kan anvendes på mange forskellige tumortyper (herunder PC-3 prostatacancer, lungecarcinom, og muse mamma fedtpude tumorer), men i dette manuskript vil vi fokusere på brystkræft model, MDA-MB-231. I denne model bruger vi en meget knogle-selektiv klon, oprindeligt stammer i Dr. Mundy gruppe i San Antonio 2, at hsom siden er blevet transficeret for GFP-ekspression og re-klonet ved vores gruppe 3. Denne klon er et knogle metastatisk variant med en høj osteotropism og meget lidt metastaser til lunge, lever, eller binyrerne. Mens intrakardiale injektioner er mest almindeligt anvendt til studier af knoglemetastaser 2, i visse tilfælde intratibial 4 eller yverfedt pad indsprøjtninger er mere passende. Intrakardiale injektioner udføres typisk ved anvendelse af humane tumorceller med det formål at overvåge senere stadier af metastaser, især evnen af ​​cancerceller til at standse i knogler, overlever, prolifererer og fastslå tumorer, der udvikler sig til cancer-induceret knoglesygdom. Intratibial injektioner foretages, hvis der fokuseres på forholdet mellem kræftceller og knogler efter en tumor har spredt til knogler, som korrelerer nogenlunde fast metastatisk knoglesygdom. Ingen af ​​disse modeller rekapitulerer tidlige trin i den metastatiske proces forud for emboli og indgangaf tumorceller til kredsløbet. Hvis overvågningen af ​​primær tumorvækst eller metastase fra det primære sted til knogler, derefter bryst-fedtpuden inokulationer foretrækkes normalt, men vil meget få tumorcellelinier konsekvent metastaserer til knogle fra det primære site, med 4T1 knogle-præferentielle kloner, en musemammatumorvirus carcinom, være undtagelsen 5,6. Dette håndskrift detaljer inokulering procedurer og fremhæver centrale skridt i efter inokulering analyser. Konkret omfatter det cellekultur, tumorcelle inokulering procedurer for intrakardiale og intratibial vacciner, samt kortfattet information om ugentlig overvågning af x-ray, fluorescens og histomorfometriske analyser.

Watch this Scientific Journal Video about Models of Bone Metastasis at JoVE.com

Models of Bone Metastasis

Dyremodeller ofte anvendes til at studere cancermetastase til knogler. I denne protokol vil vi beskrive to almindelige metoder til tumorinoculation for knoglemetastaser undersøgelser og kort beskrive nogle af de analyser, der anvendes til at overvåge og kvantificere disse modeller.

Modeller af knoglemetastaser

Bone metastases are a common occurrence in several malignancies, including breast, prostate, and lung. Once established in bone, tumors are responsible ...

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Medicine

Experimental Metastasis Assay

Metastasis is the leading cause of death in cancer patients.  To understand the mechanism of metastasis, an experimental metastasis assay was established ...

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

In vitro mesothelial Clearance Assay at modeller de tidlige stadier af kræft i æggestokkene Metastase

Ovarian cancer is the fifth leading cause of cancer related deaths in the United States1. Despite a positive initial response to therapies, 70 to 90 ...

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Multi-modal Imaging af angiogenese i en nøgen rotte model af Breast Cancer knoglemetastaser Brug Magnetic Resonance Imaging, Volumetrisk Computed Tomography og Ultralyd

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

Præklinisk evaluering af tyrosinkinasehæmmere for Behandling af akut leukæmi

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

En ortotopisk musemodel for Human Prostata Cancer metastaser

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models

Justerbar Stivhed, Eksterne fixator for Rat Femur osteotomi og Segmentoplysninger knogledefekt Modeller

The mechanical environment around the healing of broken bone is very important as it determines the way the fracture will heal. Over the past decade there ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

Højfrekvente Ultrasound Imaging af Mouse Cervikal lymfeknuder

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

En &quot;Patient-Like&quot; ortotopisk Syngene musemodel af hepatocellulært carcinom Metastase

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Kvantificering af Intra-peritoneal Kræft i æggestokkene Metastase

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Evaluation of Lung Metastasis in Mouse Mammary Tumor Models by Quantitative Real-time PCR

Evaluering af lungemetastase i musebrysttumor modeller af kvantitativ real-time PCR

Metastatic disease is the spread of malignant tumor cells from the primary cancer site to a distant organ and is the primary cause of cancer associated ...