Name: establishing dual resistance to egfr-tki and met-tki in lung adenocarcinoma cells in vitro with a 2-step dose-escalation procedure
Uploaded: 2017-08-11T14:00:00
Description: Watch this Scientific Journal Video about Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure at JoVE.com

Vi takke medlemmerne af Institut for Molekylær onkologi for deres tankevækkende kommentarer og nyttige råd, og Editage for deres hjælp med engelsksprogede redigering. Dette arbejde blev støttet af den MEXT-støttet Program for den strategiske forskningsfond på Private universiteter (2012-2016) fra ministeriet for uddannelse, kultur, sport, videnskab og teknologi i Japan. Tohru Ohmori modtaget forskningsstøtte (2015-2016) fra Boehringer-Ingelheim (Ingelheim, Tyskland).

1. fastlægge Gefitinib-resistente celler
<ol>
	<li>Bestemme den oprindelige gefitinib koncentration af 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromid (MTT) assay.
	<ol>
		<li>Kultur PC-9 celler i vækstmediet (RPMI-1640 medium med 10% FBS og 1% antibiotika (penicillin: 100 U/mL og streptomycin: 100 µg/mL)), i en 5% CO2 inkubator ved 37 ° C.</li>
		<li>Justere celler til 1,0 × 105 celler/mL ved hjælp af en automatiseret celle counter (Se tabel over materia...

<ol>
	<li>Lynch, T. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activating+mutations+in+the+epidermal+growth+factor+receptor+underlying+responsiveness+of+non-small-cell+lung+cancer+to+gefitinib.">Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib.</a> N Engl J Med. 350 (21), 2129-2139 (2004).</li><li>Paez, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EGFR+mutations+in+lung+cancer:+correlation+with+clinical+response+to+gefitinib+therapy.">EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy.</a> Science. 304 (5676), 1497-1500 (2004).</li><li>Pao, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EGF+receptor+gene+mutations+are+common+in+lung+cancers+from+"never+smokers"+and+are+associated+with+sensitivity+of+tumors+to+gefitinib+and+erlotinib.">EGF receptor gene mutations are common in lung cancers from "never smokers" and are associated with sensitivity of tumors to gefitinib and erlotinib.</a> Proc Natl Acad Sci U S A. 101 (36), 13306-13311 (2004).</li><li>Mitsudomi, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+of+the+epidermal+growth+factor+receptor+gene+predict+prolonged+survival+after+gefitinib+treatment+in+patients+with+non-small-cell+lung+cancer+with+postoperative+recurrence.">Mutations of the epidermal growth factor receptor gene predict prolonged survival after gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence.</a> J Clin Oncol. 23 (11), 2513-2520 (2005).</li><li>Sequist, L. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotypic+and+histological+evolution+of+lung+cancers+acquiring+resistance+to+EGFR+inhibitors.">Genotypic and histological evolution of lung cancers acquiring resistance to EGFR inhibitors.</a> Sci Transl Med. 3 (75), 75ra26 (2011).</li><li>Engelman, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MET+amplification+leads+to+gefitinib+resistance+in+lung+cancer+by+activating+ERBB3+signaling.">MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling.</a> Science. 316 (5827), 1039-1043 (2007).</li><li>Schmidt, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+mutations+of+the+MET+proto-oncogene+in+papillary+renal+carcinomas.">Novel mutations of the MET proto-oncogene in papillary renal carcinomas.</a> Oncogene. 18 (14), 2343-2350 (1999).</li><li>Olivero, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+mutation+in+the+ATP-binding+site+of+the+MET+oncogene+tyrosine+kinase+in+a+HPRCC+family.">Novel mutation in the ATP-binding site of the MET oncogene tyrosine kinase in a HPRCC family.</a> Int J Cancer. 82 (5), 640-643 (1999).</li><li>Janne, P. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AZD9291+in+EGFR+inhibitor-resistant+non-small-cell+lung+cancer.">AZD9291 in EGFR inhibitor-resistant non-small-cell lung cancer.</a> N Engl J Med. 372 (18), 1689-1699 (2015).</li><li>Bean, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MET+amplification+occurs+with+or+without+T790M+mutations+in+EGFR+mutant+lung+tumors+with+acquired+resistance+to+gefitinib+or+erlotinib.">MET amplification occurs with or without T790M mutations in EGFR mutant lung tumors with acquired resistance to gefitinib or erlotinib.</a> Proc Natl Acad Sci U S A. 104 (52), 20932-20937 (2007).</li><li>. NCT02468661 - ClinicalTrials.gov Available from: <a target="_blank" href="https://clinicaltrials.gov/ct2/show/NCT02468661">https://clinicaltrials.gov/ct2/show/NCT02468661</a> (2017)</li><li>Yamaoka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+Resistance+Mechanisms+to+Combination+Met-TKI/EGFR-TKI+Exposure+in+Met-Amplified+EGFR-TKI-Resistant+Lung+Adenocarcinoma+Harboring+an+Activating+EGFR+Mutation.">Acquired Resistance Mechanisms to Combination Met-TKI/EGFR-TKI Exposure in Met-Amplified EGFR-TKI-Resistant Lung Adenocarcinoma Harboring an Activating EGFR Mutation.</a> Mol Cancer Ther. 15 (12), 3040-3054 (2016).</li><li>Mosmann, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+colorimetric+assay+for+cellular+growth+and+survival:+application+to+proliferation+and+cytotoxicity+assays.">Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.</a> J Immunol Methods. 65 (1-2), 55-63 (1983).</li><li>Cifone, M. A., Fidler, I. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlation+of+patterns+of+anchorage-independent+growth+with+in+vivo+behavior+of+cells+from+a+murine+fibrosarcoma.">Correlation of patterns of anchorage-independent growth with in vivo behavior of cells from a murine fibrosarcoma.</a> Proc Natl Acad Sci U S A. 77 (2), 1039-1043 (1980).</li><li>Ercan, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amplification+of+EGFR+T790M+causes+resistance+to+an+irreversible+EGFR+inhibitor.">Amplification of EGFR T790M causes resistance to an irreversible EGFR inhibitor.</a> Oncogene. 29 (16), 2346-2356 (2010).</li><li>Ogino, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emergence+of+epidermal+growth+factor+receptor+T790M+mutation+during+chronic+exposure+to+gefitinib+in+a+non+small+cell+lung+cancer+cell+line.">Emergence of epidermal growth factor receptor T790M mutation during chronic exposure to gefitinib in a non small cell lung cancer cell line.</a> Cancer Res. 67 (16), 7807-7814 (2007).</li><li>Ando, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+sensitivity+to+tumor+necrosis+factor+alpha+in+non-small+cell+lung+cancer+cells+with+acquired+resistance+to+gefitinib.">Enhancement of sensitivity to tumor necrosis factor alpha in non-small cell lung cancer cells with acquired resistance to gefitinib.</a> Clin Cancer Res. 11 (24 Pt 1), 8872-8879 (2005).</li><li>Suda, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reciprocal+and+complementary+role+of+MET+amplification+and+EGFR+T790M+mutation+in+acquired+resistance+to+kinase+inhibitors+in+lung+cancer.">Reciprocal and complementary role of MET amplification and EGFR T790M mutation in acquired resistance to kinase inhibitors in lung cancer.</a> Clin Cancer Res. 16 (22), 5489-5498 (2010).</li><li>Shien, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+resistance+to+EGFR+inhibitors+is+associated+with+a+manifestation+of+stem+cell-like+properties+in+cancer+cells.">Acquired resistance to EGFR inhibitors is associated with a manifestation of stem cell-like properties in cancer cells.</a> Cancer Res. 73 (10), 3051-3061 (2013).</li><li>Sagawa, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+three+cisplatin-resistant+endometrial+cancer+cell+lines+using+two+methods+of+cisplatin+exposure.">Establishment of three cisplatin-resistant endometrial cancer cell lines using two methods of cisplatin exposure.</a> Tumour Biol. 32 (2), 399-408 (2011).</li><li>Ritter, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+breast+cancer+cells+selected+for+resistance+to+trastuzumab+in+vivo+overexpress+epidermal+growth+factor+receptor+and+ErbB+ligands+and+remain+dependent+on+the+ErbB+receptor+network.">Human breast cancer cells selected for resistance to trastuzumab in vivo overexpress epidermal growth factor receptor and ErbB ligands and remain dependent on the ErbB receptor network.</a> Clin Cancer Res. 13 (16), 4909-4919 (2007).</li></ol>

Protokollen beskrevet her kan bruges til oprettelse af erhvervede dual-modstand i kræftceller ved hjælp af en 2-trins dosis-eskalering metode in vitro. Mange forskningsartikler har rapporteret, at PC-9 celler udviklet EGFR-TKI (gefitinib eller erlotinib) modstand gennem T790M EGFR mutationer15,16. Men vi gjorde ikke opdager T790M mutationer i exon 20 af vores PC-9MET1000 celler af direkte sekventering (figur 4 c). Derudove...

Muterede mutationer i epidermal vækstfaktor receptor (EGFR) er fundet i en delmængde af patienter med ikke-småcellet lungekræft (NSCLC) og er vigtige prædiktive biomarkører for denne sygdom1,2. Disse aktivering EGFR mutationer oftest forekomme som en i-frame sletning i exon 19 (delE746-A750) eller et punkt mutation erstatter leucin med arginin på codon 858 af exon 21 (L858R) i EGFR tyrosin kinase domæne3,4. Behandling med EGFR-tyrosin kinase hæmmere (TKIs) er en klinisk effektiv behandling for patienter med NSCLC husly EGFR mutationer. Men, denne terapi er begrænset af den uundgåelige erhvervelse af modstand mod EGFR-TKIs som gefitinib og erlotinib. Den mest almindelige erhvervet resistens opstår gennem en sekundær T790M mutation i exon 20 af EGFR, som blev fundet i ca. 60% af patienter, der har erhvervet EGFR-TKI (gefitinib eller erlotinib) modstand. Andre molekylære mekanismer forbundet med erhvervet modstand mod EGFR-TKIs er aktivering af bypass signalering forårsaget af markedsøkonomisk forstærkning, transformation af små-cellet lungekræft og epitelial til mesenchymale overgangen5,6. Receptoren for den hepatocyt vækst faktor, kodet af genet MET som er dysregulatedin mange tumorer, er blevet rapporteret til at være en vigtig kandidat til målrettede behandlinger7,8.
Strategier for at overvinde den erhvervede modstand mod EGFR-TKIs er nu under klinisk evaluering. Prækliniske og kliniske undersøgelser har vist, at tredje generation EGFR-hæmmere såsom osimertinib er effektive til patienter med T790M mutation9. Væksten i EGFR-mutant kræftformer med markedsøkonomisk forstærkning kan desuden hæmmes af kombineret behandling med EGFR - og markedsøkonomisk-TKIs6,10. Kliniske forsøg vurderer en kombination af markedsøkonomisk behandling og EGFR-hæmmere hos patienter med erhvervet modstand mod gefitinib og erlotinib er igangværende nu11.
For at studere den molekylære mekanisme af erhvervet modstand mod kemoterapeutika, behandles cellelinjer, der i første omgang reagerer på hæmmere løbende med disse hæmmere. Der findes to metoder til at etablere erhvervet modstand i cellelinjer. Ene er trinvis dosis-eskalering, og den anden er høj koncentration eksponering. Trinvis eskalering metode er blevet mere almindeligt brugt, fordi det har en højere succesrate. Cellerne er først udsat for lave koncentrationer af hæmmere, næsten en tiendedel af værdien halv-maksimal hæmmende koncentration (IC50), og koncentrationen er gradvist derefter steg med 10-30% indtil målet koncentration opnås. Høj koncentration eksponering metode indebærer at udsætte celler til den sidste dosis af inhibitor dyrkningsmedium, hvilket er mere end værdien IC50 så forældrenes cellerne dræbes næsten helt. Denne høj koncentration eksponering metode har meget en lavere succesrate end metoden trinvis eskalering.
I en tidligere undersøgelse, blev kombineret behandling med EGFR-TKI og markedsøkonomisk-TKI vist sig at være effektiv i en in vitro- system. Erhvervet modstand mod en EFGR-TKI, gefitinib, blev etableret gennem gradvis optrapning i 12 måneder, sammen med MET-forstærkning, og således en gefitinib-resistente cellelinje kaldet PC-9MET1000 blev genereret12.
Protokollen præsenteres her er en totrinsprocedure på dosis-eskalering søges EGFR-muteret NSCLC PC-9 celler med dobbelt modstand mod EGFR-TKI, gefitinib og en MET-TKI, PHA665752 (figur 1). Denne metode til oprettelse af erhvervet resistente i cellelinjer er relativt nemme at udføre, med en høj succesrate. Den beskrevne protokol kan ændres for anvendelse med hæmmere af andre Molekylær målet narkotika og cytotoksiske stoffer samt.

<table width="970">
	<tbody>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">RPMI-1640</td>
			<td style="width:177px;">Wako</td>
			<td style="width:119px;">189-02025</td>
			<td style="width:309px;">with L-Glutamine and Phenol Red</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CellTiter 96</td>
			<td style="width:177px;">Promega</td>
			<td style="width:119px;">G4100</td>
			<td>Non-Radioactive Cell Proliferation Assay</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">FBS</td>
			<td style="width:177px;">gibco</td>
			<td style="width:119px;">26140-079</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">Pen Strep</td>
			<td style="width:177px;">gibco</td>
			<td style="width:119px;">15140-163</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">gefitinib</td>
			<td style="width:177px;">Selleck</td>
			<td style="width:119px;">S1025</td>
			<td>stocked 10 mM/EMSO at -20&#186;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">PHA665752</td>
			<td style="width:177px;">Selleck</td>
			<td style="width:119px;">S1070</td>
			<td>stocked 10 mM/EMSO at -20&#186;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">96-well plate</td>
			<td style="width:177px;">IWAKI</td>
			<td style="width:119px;">860-096</td>
			<td>flat bottom with lid, low evaporation type</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">TC10 Automated Cell Counter</td>
			<td style="width:177px;">BIO RAD</td>
			<td>#1450010</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="width:365px;height:26px;">CellTiter 96 Non-Radioactive Cell Proliferation Assay</td>
			<td style="width:177px;">Promega</td>
			<td style="width:119px;">G4100</td>
			<td style="width:309px;">Dye Solution and Solubilization/Stop Solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Powerscan HT microplate reader</td>
			<td>BioTek</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="width:365px;height:26px;">GraphPad Prism v.5.0 software</td>
			<td style="width:177px;">GraphPad Inc.</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="width:365px;height:26px;">PC-9</td>
			<td style="width:177px;">Riken BioResource Center</td>
			<td style="width:119px;">RCB4455</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">P100 dish</td>
			<td style="width:177px;">FALCON</td>
			<td align="right" style="width:119px;">353003</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishing dual resistance to egfr-tki and met-tki in lung adenocarcinoma cells in vitro with a 2-step dose-escalation procedure

Resistens udgør en stor udfordring i kræftbehandling. Generation af resistente sublines in vitro- er nødvendige for at opdage nye mekanismer til at overvinde denne udfordring. Her, en 2-trins dosis-eskalering metode til fastlæggelse af dual-modstand mod en epidermal vækstfaktor receptor (EGFR)-tyrosin kinase inhibitor (TKI), gefitinib og en MET-TKI, PHA665752, er beskrevet. Denne metode er baseret på enkle trinvis dosis-eskalering af inhibitorer for inducerende erhvervet modstand i cellelinjer. Den alternative metode til generering af resistente sublines indebærer udsætter celler for høje koncentrationer af inhibitor i ét trin. Metoden trinvis dosis-eskalering har en højere mulighed for med held inducerende erhvervet modstand end denne metode. Aktivering EGFR er mutationer biomarkører for en reaktion på behandling med EGFR-TKI, som er en anvendt første linje behandling af ikke-småcellet lungekræft (NSCLC), at havnen disse mutationer. Men trods rapporter om effektive svar, brug af EGFR-TKI er begrænset, fordi tumorer uundgåeligt erhverve modstand. De vigtigste mekanismer bag EGFR-TKI modstand omfatte en sekundær mutation på webstedet gatekeeper, T790M i exon 20 af EGFR og en bypass signal om markedsøkonomisk behandling. En mulig løsning på problemet vil således være en kombination af EGFR-TKI og markedsøkonomisk-TKI. Denne kombinerede behandling har vist sig at være effektiv i en in vitro- undersøgelse model. Erhvervede gefitinib-modstand blev etableret gennem MET-forstærkning af trinvis dosis-eskalering af gefitinib i 12 måneder, og en cellelinje opkaldt PC-9MET1000 blev genereret i en tidligere undersøgelse. For at yderligere undersøge mekanismerne af erhvervede MET-TKI og EGFR-TKI blev modstand, en MET-TKI, PHA665752, administreret til disse celler med trinvis dosis-eskalering i overværelse af gefitinib i 12 måneder. Denne protokol har også været anvendt med succes for en række Kombinationsbehandlinger at etablere erhvervet resistens over for andre hæmmer molekyler.

Oversigt over protokollen præsenteret i dette håndskrift er vist i figur 1, inklusive induktion af erhvervede dual-modstand mod gefitinib og PHA665752 i PC-9 celler af en 2-trins dosis-eskalering procedure. Figur 2 viser et fald i udbredelsen af forældrenes PC-9 celler som koncentrationen af gefitinib stigninger. Dette indikerer, at PC-9 celler blev følsomme over for gefitinib efter proceduren dosis-eskalering blev udført. <...

Watch this Scientific Journal Video about Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure at JoVE.com

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

En in vitro- metode til fastlæggelse af dobbelt modstand mod en EGFR-TKI og en MET-TKI i kræftceller er beskrevet. Denne metode er nyttig for at udvikle behandlinger til patienter med EGFR-mutationer, som udviser sygdomsprogression trods EGFR-TKI behandling med MET-forstærkning. Det kan også ændres for hæmmere rettet mod andre molekyler.

Oprettelse af dobbelt modstand mod EGFR-TKI og MØDTE TKI i lunge adenocarcinom celler In Vitro med en 2-trins dosis-eskalering Procedure

Drug resistance is a major challenge in cancer therapy. The generation of resistant sublines in vitro is necessary for discovering novel mechanisms to ...

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Cancer Research

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

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Etablering og karakterisering af tre Afatinib-resistente lunge adenokarcinom PC-9 cellelinjer udviklet med stigende doser Afatinib

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Tumor associated macrophages (TAMs) account for a large percentage of cells in the tumor mass for different types of cancers. Glioblastoma (GBM), a ...

In Vitro Evaluation of Oncogenic Transformation in Human Mammary Epithelial Cells

In Vitro evaluering af onkogene transformation i humane mammary epitelceller

Tumorigenesis is a multi-step process in which cells acquire capabilities&#160;that allow their growth, survival, and dissemination under hostile ...

Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model

Brug computer-baseret billedanalyse til at forbedre kvantificeringen af lungemetastase i 4T1 Breast Cancer Model

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. ...

Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo

Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo

Kemisk bøjning af et renset DEC-205-rettet antistof med protein i fuld længde til målretning af musededritiske celler In Vitro og In Vivo

Targeted antigen delivery to cross-presenting dendritic cells (DC) in vivo efficiently induces T effector cell responses and displays a valuable approach ...