Name: synthesis and characterization of an aspirin-fumarate prodrug that inhibits nf&#954;b activity and breast cancer stem cells
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Description: Watch this Scientific Journal Video about Synthesis and Characterization of an Aspirin-fumarate Prodrug that Inhibits NFÎºB Activity and Breast Cancer Stem Cells at JoVE.com

This work was supported by grants provided by the National Institutes of Health (NIH), R01 CA200669 to JF and R01 CA121107 to GRJT, and by a postdoctoral fellowship grant from Susan G. Komen for the Cure to IK (PDF12229484).

Bir miktar aspirin-fumarat Ön ilaç GTCpFE 1. sentezi <ol><li> metanol, 0.81 ml (20 mmol) ölçülmesi ve yuvarlak dipli bir şişeye (10 mi) su içinde karıştırarak, plastik bir piston şırınga kullanılarak. bir buz-su banyosu içinde balon yerleştirilmesi ile 0 ° C&#39;ye kadar elde edilen karışımın soğutun. 4-hidroksibenzil alkol (2.48 mg, 20 mmol) ilave edilmiş ve çözelti berrak olana kadar, reaksiyon karışımı karıştırılmıştır. </li><li> O -acetylsalicyloyl klorür, istenen mik...

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In this protocol, we demonstrated the synthesis of an ASA prodrug, GTCpFE, where the fumarate pharmacophore was incorporated to improve the anti-NF&#312;B activity in breast cancer cells. GTCpFE is an effective NF&#312;B inhibitor, whereas ASA itself is not, even at much higher concentrations. The fumarate moiety has anti-inflammatory properties as shown by its ability to inhibit NF&#312;B signaling in a variety of cell lines and tissues14,25-29. The prodrug strategy described herein, is amendable to other mal...

Enflamasyon böyle metastazı 1&#39;e insidansı ve tanıtım, ve sonunda ilerleme olarak tümörogenez birden yönlerini altında yatan bir özelliğidir. Meme kanserinde, bu daha da metastaz ve nüks hem meme kanseri insidansı azalma ve riski ile ilişkilidir (salisilik asit, ASA asetil) klasik non-steroid anti-inflamatuar ilaç aspirin o düzenli kullanımını gösteren epidemiyolojik gözlemlerle desteklenmektedir 2,3. ASA esas Meme kanseri 4,5 yukarı regüle edilir, siklooksijenaz-2 aktivitesini inhibe ederek etki eder. Ancak, ASA anti-kanser etkileri de anormal nükleer faktör KB&#39;yi (NFKB) 6-8 sinyal bastırılması aracılık edebilir. Bir kuralsız NFKB yolu tedavisine 9-11 tümör hücre sağkalım, çoğalması, göç, işgali, anjiogenezisi ve direncini arttırır, çünkü bu önemlidir. NFKB yolu aktivasyonu da monte edilmesi için çok önemlidirBir bağışıklık tepkisi. Bu nedenle, uzun süreli NFKB inhibisyonunun gerekli olduğu bir anti-kanser terapisi, bir uzun süreli bir bağışıklık bastırma kapsayan zararlı yan etkileri göz önünde bulundurulmalıdır. Bu nedenle, ASA tedavi optimizasyonu için iyi bir başlangıç ​​noktası olarak hizmet verebilir. Kanser terapisinde ASA uygulama için bir sınırlama örneğin ülser ve mide 12,13 kanama gibi, mide-bağırsak toksisitesi ile bağlantılı olan siklooksijenaz 2 ve NFKB inhibisyonu için gerekli olan yükseltilmiş dozlar vardır. Ancak, ester ön ilacı haline ASA dönüştürerek, ASA gastrointestinal toksisite azaltabilir. ayrıca gücünü artırmak ve / veya işlevsellik eklemek için, ek yapı elemanları veya yardımcı farmakoforlarına da ester ön tasarımına dahil edilebilir. Bu tür bir farmakofor NFKB yolu fumarattır karşı daha önce NFKB yolu inhibisyonu 14,15 için önemli olduğu gösterilmiştir ki, ASA gücünü arttırmak için ilave edildi. 15, GTCpFE ve bu hibrid molekül henüz NFKB yolunun karşı güçlü güvenli olacağını varsaydık. Meme kanseri hücreleri içinde, anti-NFKB aktivitesini ve NFKB yaşama ve büyüme 16-21 için sinyal kullanan meme kanseri kök hücreleri (rulmanları) 15, bloke etme kabiliyetini test etmiştir. Biz NFKB yolunun karşı GTCpFE gücü önemli ölçüde ASA 15 üzerinde geliştirilmiş olduğunu bulmak. Buna ek olarak, GTCpFE blok mammosphere oluşumu ve CSC yüzey işaretleyici CD44 + CD24 azaltır - GTCpFE CSCS 15 yok etme kapasitesine sahip olduğunu gösteren, immünfenotip. Bu sonuçlar, aynı zamanda, meme CSCS hedefleyebilir etkili bir anti-enflamatuvar ajan olarak aspirin fumarat ön-ilaç oluşturmak. Meme kanseri tedavisi açısından, GTCpFE agresif ve ölümcül hastalığın tedavisinde bir potansiyele sahip olabilir.

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			<td height="33" style="height:33px;width:216px;">RPMI 1640 Red Medium</td>
			<td style="width:123px;">Life Technologies&#160;</td>
			<td style="width:119px;">11875-093</td>
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			<td height="33" style="height:33px;width:216px;">IMEM</td>
			<td style="width:123px;">Corning</td>
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			<td height="33" style="height:33px;width:216px;">DMEM / F12 Medium</td>
			<td style="width:123px;">ThermoFisher</td>
			<td style="width:119px;">21041-025</td>
			<td style="width:205px;">Warm up to 37&#176;C before use</td>
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			<td height="33" style="height:33px;width:216px;">MEM NEAA 10mM 100X</td>
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			<td height="33" style="height:33px;width:216px;">Penicillin Streptomycin</td>
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			<td height="33" style="height:33px;width:216px;">Insulin</td>
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			<td style="width:119px;">I-1882</td>
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		<tr height="33">
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			<td style="width:205px;"></td>
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		<tr height="33">
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			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;">NF&#954;B-RE Luciferase Construct&#160;</td>
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			<td height="33" style="height:33px;">CyAn ADP Analyzer&#160;</td>
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			<td height="33" style="height:33px;">QCapture Software</td>
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			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;">Summit Software</td>
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			<td height="33" style="height:33px;">GraphPad Software</td>
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			<td style="width:119px;"></td>
			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;width:216px;">TRIzol</td>
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			<td height="33" style="height:33px;width:216px;">M-MLV Reverse Transcriptase</td>
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			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;width:216px;">100 mM dNTP Set</td>
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			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;width:216px;">Random Hexamers&#160;</td>
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			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;width:216px;">Fast SYBR Green Master Mix</td>
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			<td></td>
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		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Costar 96W, ultra low attachment&#160;</td>
			<td style="width:123px;">Corning</td>
			<td style="width:119px;">3474</td>
			<td></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">HBSS, 1X</td>
			<td style="width:123px;">ThermoFisher</td>
			<td style="width:119px;">14025134</td>
			<td></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">CD44-APC conjugated antibody&#160;</td>
			<td style="width:123px;">BD Pharmingen</td>
			<td style="width:119px;">560990</td>
			<td style="width:205px;">Store at 4&#176;C, protect from light</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">CD24-PE antibody</td>
			<td style="width:123px;">BD Pharmingen</td>
			<td style="width:119px;">560991</td>
			<td style="width:205px;">Store at 4&#176;C, protect from light</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Recombinant Human TNF&#945;</td>
			<td style="width:123px;">Fisher</td>
			<td style="width:119px;">210TA100&#160;</td>
			<td></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">CCL2 Primer Forward Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">AGAATCACCAGCAGCAAGTGTCC</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">CCL2 Primer Reverse Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">TCCTGAACCCACTTCTGCTTGG</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">ICAM1 Primer Reverse Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">TGACGAAGCCAGAGGTCTCAG</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">ICAM1 Primer Forward Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">AGCGTCACCTTGGCTCTAGG</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">TNF Primer Forward Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">AAGGGTGACCGACTCAGCG</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">TNF Primer Reverse Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">ATCCCAAAGTAGACCTGCCCA</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">36B4 Primer Forward Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">GTGTTCGACAATGGCAGCAT</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">36B4 Primer Reverse Sequence</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">GACACCCTCCAGGAAGCGA</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Sodium Hydroxide</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">S5881-500G</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">4-Hydroxybenzyl Alcohol</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">20806-10G</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">O-Acetylsalicyloyl Chloride</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">165190-5G</td>
			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;width:216px;">Phosphorous Pentoxide</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">79610-100G</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Ethyl Fumaroyl Chloride</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">669695-1G</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Sodium Sulfate</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">246980-500G</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">4-Dimethylaminopyridine</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">714844-100ML</td>
			<td style="width:205px;">0.5&#160;M in tetrahydrofuran</td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Triethylamine</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">T0886-100ML</td>
			<td style="width:205px;"></td>
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			<td height="33" style="height:33px;width:216px;">Toluene</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">244511-100ML</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Ethyl Acetate</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">270989-100ML</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:33px;width:216px;">Tetrahydrofuran</td>
			<td style="width:123px;">Sigma-Aldrich</td>
			<td style="width:119px;">401757-2L</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:216px;">400 MHz FT NMR spectrometer&#160;</td>
			<td style="width:123px;"></td>
			<td style="width:119px;"></td>
			<td style="width:205px;">See Bruker&#8217;s Avance User&#8217;s Manual, version 000822 for details</td>
		</tr>
	</tbody>
</table>

synthesis and characterization of an aspirin-fumarate prodrug that inhibits nf&#954;b activity and breast cancer stem cells

Enflamasyon metastazı kanser insidansını ve tanıtım, ve sonunda ilerlemesini altında yatan bir kanser özelliğidir. Bu nedenle, standart kanser alaylara karşı bir anti-inflamatuar ilaç eklenmesi hasta sonuçlarını artırabilir. Bu tür bir madde, aspirin (asetilsalisilik asit, ASA), kanserinin kimyasal yollardan önlenmesine ve anti-tümör aktivitesi için incelenmiştir. siklooksijenaz 2-prostaglandin ekseni inhibe yanı sıra, ASA anti-kanser aktiviteleri, aynı zamanda nükleer faktör ĸB (NFĸB) inhibisyonuna bağlanmıştır. Uzun süreli ASA kullanımının sindirim toksisiteye neden olabilir, çünkü bir ön stratejisi başarıyla uygulanmaktadır. Bu ön ilacın içinde ASA karboksilik asit maskeli ve ek farmakoforlarına dahil edilir tasarımı. Bu protokol, biz aspirin fumarat ön ilacı GTCpFE sentezlendi ve meme kanseri hücrelerinde NFĸB yolunun önlemesini, özelliği, kanser zayıflaması mil benzeri uygun açıklamaktadırkravat, önemli bir NFĸB bağımlı fenotip. ASA ASA yapısına eklenmesi fumarat önemli ölçüde etkinliğine katkıda bulunduğunu belirten, herhangi bir inhibe edici aktiviteye yoksun ise GTCpFE, meme kanseri hücre hatlarında NFĸB yolu inhibite eder. Immünfenotip - Buna ek olarak, GTCpFE mammosphere oluşumunu engelleme ve kanser kök hücresi ile ilişkili CD44 + CD24 hafifleterek önemli anti-kanser kök hücre aktivitesi gösterir. Bu sonuçlar kemoprevensiyon ve kanser tedavisi için geliştirilmiş bir anti-inflamatuar ilaçlar geliştirmek için uygun bir strateji oluşturmak.

Şekil 1 &#39;de, sitokin aspirin fumarat ön-ilaç, GTCpFE, ve önleyici aktivite kimyasal yapısı, meme kanseri hücrelerinde NFĸB yolu kaynaklı gösterilmiştir. GTCpFE Hücrelerarası adezyon molekülü 1 (ICAM-1), kemokin CC Motif Ligand 2 (CCL2), ve Tümör Nekroz Faktörü (TNF) (Şekil şekilde NFĸB hedef genlerin her iki NFĸB uç noktaları NFĸB RE lusiferaz aktivitesi (Şekil 1 b) ve ekspresyonunu inhibe MCF-7 göğüs ka...

Watch this Scientific Journal Video about Synthesis and Characterization of an Aspirin-fumarate Prodrug that Inhibits NFÎºB Activity and Breast Cancer Stem Cells at JoVE.com

Synthesis and Characterization of an Aspirin-fumarate Prodrug that Inhibits NF&#954;B Activity and Breast Cancer Stem Cells

This procedure will demonstrate how we synthesized and characterized the anti-NF&#954;B and anti-cancer stem cell activity of an aspirin-fumarate prodrug.

NFKB Aktivite ve Meme Kanseri Kök Hücreler önler Sentezi ve bir Aspirin-fumarat ön-ilacının Karakterizasyonu

Inflammation is a cancer hallmark that underlies cancer incidence and promotion, and eventually progression to metastasis. Therefore, adding an ...

The overall goals of this procedure are to demonstrate how the anti-inflammatory aspirin-fumarate prodrug GTCpFE is synthesized, and how it improves anti-NFkappaB and anti-cancer stem cell activity in breast cancer cells. This method can help answer key questions in the cancer therapeutics field by showing how we can repurpose or even improve anti-inflammatory drugs to target therapy resistant breast cancer stem cells. The main advantage of this technique is that we utilize the multidisciplinary approach to obtain and characterize the improved aspirin-fumarate prodrug.Demonstrating the synthesis of the aspirin-fumarate prodrug GTCpFE will be Loruhama Delgado-Rivera, a graduate student in Dr.Gregory Thatcher's laboratory. Whereas the anti-NFkappaB activity of GTCpFE in breast cancer cells will be demonstrated by Gergana Georgieva, a pharmacy student researcher in Dr.Jonna Frasor's laboratory. Using a plastic plunger syringe, measure 0.81 milliliters of methanol and mix it in 10 milliliters of water in a round-bottom flask.Cool the resulting mixture to zero degrees Celsius by placing the flask in an ice water bath. Add 2.48 milligrams of 4-hydroxybenzyl alcohol to the reaction mixture. Stir until the solution is clear.Next, prepare a solution of O-acetylsalicyloyl chloride in anhydrous toluene by dissolving the solute in the solvent in a separate flask. Using a plastic plunger syringe add this solution to the 4-hydroxybenzyl alcohol mixture and leave the reaction stirring at zero degrees celsius. Monitor the reaction using thin-layer chromatography, or TLC, as described in the text protocol.When the reaction is complete remove the ice water bath and allow the reaction to stir at room temperature for 20 hours. Filter the precipitate using a Buchner funnel with a fritted disc of medium porosity. Then, place the solid in a scintillation vial and leave the compound in vacuo overnight in a desiccator at room temperature.Next, seal a thoroughly dried round-bottom flask with a septum and pierce it with a needle that is connected to a vacuum pump system. Inflate a balloon using argon gas and connect it to a plastic syringe with a needle. Insert the needle connected to the argon balloon through the septum sealing the round-bottom flask and remove the needle connected to the vacuum pump.Then, add 4-hydroxymethyl-phenyl ester, of 2-acetyloxybenzoic acid, 4-dimethylaminopyridine and trimethylamine to a separate flask. Dissolve the chemicals in five milliliters of anhydrous tetrahydrofuran. With a plastic syringe attached to a needle add the solution to the sealed round-bottom flask.Cool the mixture down to zero degrees Celsius by placing the flask in an ice water bath. To the previous mixture, add a solution of ethyl fumaroyl chloride and anhydrous tetrahydrofuran dropwise over a period of 10 minutes. Stir the resulting mixture for two to four hours monitoring the reaction by TLC as described in the protocol.Extract the reaction mixture by adding it to a separatory funnel along with 100 milliliters of ethyl acetate and 50 milliliters of brine. Cap the funnel and shake it vigorously. Tilting the funnel to the side of the cap, slowly open the valve to allow air to escape.Once the extraction is complete, remove the aqueous phase and repeat the extraction two more times. After removing the aqueous phase, dry the mixture by adding sodium sulfate to the organic phase until the solid does not clump up when the glassware is stirred. Using another Buchner funnel with a fritted disk filter the mixture to a round-bottom flask to remove the sodium sulfate.Then, evaporate to dryness using a rotary evaporator with the temperature set at 40 degrees celsius. Next, prepare a column with silica gel and the appropriate solvent phase. Once the compound has been added, add a layer of sodium sulfate to protect the column as the solvent is added.As the column runs, monitor the eluate by TLC. Spot every other test tube as they are collected from the column. When the product of interest dilutes, mix all the samples containing pure product into a large round-bottom flask.Dry the products using a rotary evaporator with the bath temperature set at 40 degrees celsius. Perform cell culture and transfect the cells with DNA for the NFkappaB response element luciferase reporter assay as described in the text protocol. Then dissolve drug stock solutions of the GTCpFE or aspirin drugs in dimethyl sulfoxide at 1000x concentration.After 16 hours of transfection and 1 microliter of vehicle or different drug stock solutions to each well. After adding the drugs, incubate the cells for two hours at 37 degrees Celsius. To activate the NFkappaB pathway, at the proinflammatory cytokine, TNFalpha, into each well for a final concentration of 10 nanograms per milliliter.Also include a TNFalpha alone control. After incubating the cells for four hours at 37 degrees Celsius, aspirate the medium. Store the cells at minus 80 degrees Celsius.Measure luciferase using a luciferase reporter assay system according the manufacturer's instructions. To perform the mammosphere assay, first prepare mammosphere medium as described in the text protocol. Next prepare singles cells of the MDA-MB-231 cell line by trypsin digestion of monolayer cultures and filter through mesh sieves.After manually counting the single dissociated cells, plate them in 96 well ultra low attachment plates at a density of 400 cells per well. Then place the cells in the incubator at 37 degrees Celsius overnight. The next day, add different concentrations of GTCpFE to a final volume of 100 microliters.Conduct every treatment in triplicate. After seven days of culture in the incubator at 37 degrees Celsius, acquire images via imaging software using an inverted microscope. Manually count the number of mammospheres greater than 75 microns in diameter.To measure the CD44 high, CD24 low cancer stem cell immunophenotype, trypsonize MDA-MB-231 cells with 0.25%trypsin for five minutes at 37 degrees Celsius. After counting the cells using a hemocytometer, seed the cells at three million cells per dish in 10 milliliter of medium as described in the text protocol. Then add vehicle or GTCpFE to the cells and incubate at 37 degrees Celsius for 72 hours.Following incubation trypsinize the cells and distribute one million trypsinized cells in five milliliter polystyrene tubes. The tubes should contain two milliliters of 1x HBSS buffer supplemented with 2%FBS and be labeled as test or control. To stain for the surface marker CD44 and CD24, spin the cells down, aspirate the buffer, then add 20 microliters of each conjugated antibody and 80 microliters of the HBSS buffer to test tubes for a final volume of 100 microliters.Next, add 100 microliters of the same buffer to the control tubes. Also include CD44 APC-conjugated antibody and CD24 PE antibody single stained contols or the ITG immuno isotype controls. Incubate the cells in the dark at four degrees Celsius for 30 minutes.Spin down the cells for five minutes at 400 times g and reconstitute them in 200 microliters of buffer. Keep the cells on ice and in the dark. FACS analysis can now be performed.GTCpFE inhibits NFkappaB-RE luciferase activity and expression of NFkappaB target genes, such as Intercellular Adhesion Molecule 1, Chemokine C-C Motif Ligand 2, and Tumor Necrosis Factor in MCF-7 breast cancer cells. The calculated inhibitory concentration at 50%of both endpoints is approximately 20 micromolar GTCpFE. By comparison, 200 micromolar aspirin shows no inhibitory activity in breast cancer cells.This indicates that the prodrug strategy of adding the Fumarate pharmacore to aspirin significantly improves it's anti-NFkappaB activity. GTCpFE inhibits mammosphere formation of MDA-MB-231 breast cancer cells in a dose dependent manner. Similar to NFkappaB pathway inhibition in adherent cultures, the IC50 value for mammosphere formation is approximately 20 micromolar.Also measured was the population of cells expressing the CD44 high, CD24 low immunophenotype which is a bonafide cancer stem cell surface marker in breast cancer. GTCpFE pretreament resulted in a significant depletion of the CD44 high, CD24 low population in MDA-MB-231 cells. Together these results establish GTCpFE's ability to effectively target breast cancer stem cells.After watching this video, you should have a good understanding of how the design and synthesize in aspirin fumarate prodrug and how to characterize its anti-NFkappaB and anti-cancer stem cell activity on breast cancer cells. We first had the idea for this method when aspirin failed to inhibit the NFkappaB pathway in breast cancer cells as it was previously reported in the literature. The implications of this technique extend toward anti-cancer stem cell therapy, because we showed that by targeting multiple proinflammatory pathways we can in fact, effectively eradicate breast cancer stem cells.Although this method can provide insight into how to design and synthesize and characterize anti-inflammatory agents in breast cancer, this can be applied to other malignancies where multiple proinflammatory pathways are active and contribute to the pathology of the disease. Following this procedure, other methods such as the aldefluor assay and in vivo tumorigenicity can be utilized in order to asses the effect of the drug on breast cancer stem cells. The technique can pave the way for researchers in cancer therapeutics to explore prodrug design of antiinflammatory agents.GTCpFE may serve as a prototype for developing new fumarate and aspirin based antiinflammatory agents. And this could also be the basis of a new class of anti-cancer stem cell agents.

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Sentez ve karakterizasyonu plasental kondroitin sülfat (plCSA) - Lipid hedefleme - polimer nano tanecikleri

An effective cancer therapeutic method reduces and eliminates tumors with minimal systemic toxicity. Actively targeting nanoparticles offer a promising ...

Flow Cytometric Detection of Newly-formed Breast Cancer Stem Cell-like Cells After Apoptosis Reversal

Apoptozis ters sonra yeni kurulan meme kanser kök hücre benzeri hücre akış sitometrik algılama

Cancer recurrence has long been studied by oncologists while the underlying mechanisms remain unclear. Recently, we and others found that a phenomenon ...

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

Üç Afatinib dirençli akciğer adenokarsinoma PC-9 hücre hatları Afatinib artan dozlarda ile geliştirilen kurulması ve karakterizasyonu

Acquired resistance to molecular target inhibitors is a severe problem in cancer therapy. Lung cancer remains the leading cause of cancer-related death in ...

Spatial and Temporal Control of Murine Melanoma Initiation from Mutant Melanocyte Stem Cells

Mutant melanosit kök hücrelerinden Murine Melanoma başlangıcı uzamsal ve temporal kontrol

Cutaneous melanoma is well known as the most aggressive skin cancer. Although the risk factors and major genetic alterations continue to be documented ...

Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors

Jinekolojik ve Meme Kanseri Tümörlerinden Kanser Kök Hücre Kürelerinin Alınması

Cancer stem cells (CSC) are a small population with self-renewal and plasticity which are responsible for tumorigenesis, resistance to treatment and ...

Radiosensitivity of Cancer Stem Cells in Lung Cancer Cell Lines

Akciğer Kanseri Hücre Hatlarında Kanser Kök Hücrelerinin Radyosensitivite

The presence of cancer stem cells (CSCs) has been associated with relapse or poor outcomes after radiotherapy. Studying radioresistant CSCs may provide ...

Differentiation of Mouse Breast Epithelial HC11 and EpH4 Cells

Fare Meme Epitel HC11 ve EpH4 Hücrelerinin Farklılaşması

Cadherins play an important role in the regulation of cell differentiation as well as neoplasia. Here we describe the origins and methods of the induction ...

Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model

Ortotopik Meme Kanseri Modeli Kullanarak Üçlü Negatif Meme Kanseri Eğitimi

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less ...

Orthotopic Transplantation of Breast Tumors as Preclinical Models for Breast Cancer

Meme Kanseri için Preklinik Modeller Olarak Meme Tümörlerinin Ortopedik Transplantasyonu

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. ...