<meta charset="utf8"></head><body>Yeni Demir Şelatörlerinin Tespiti için Floresan Testi

<ol>
	<li>Hsu, P. P., Sabatini, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+cell+metabolism:+Warburg+and+beyond.">Cancer cell metabolism: Warburg and beyond.</a> Cell. 134 (5), 703-707 (2008).</li><li>Hsu, M. Y., Mina, E., Roetto, A., Porporato, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron:+An+essential+element+of+cancer+metabolism.">Iron: An essential element of cancer metabolism.</a> Cells. 9 (12), 2591 (2020).</li><li>Torti, S. V., Torti, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+and+cancer:+More+ore+to+be+mined.">Iron and cancer: More ore to be mined.</a> Nat Rev Cancer. 13 (5), 342-355 (2013).</li><li>Zhang, C., Zhang, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+homeostasis+and+tumorigenesis:+Molecular+mechanisms+and+therapeutic+opportunities.">Iron homeostasis and tumorigenesis: Molecular mechanisms and therapeutic opportunities.</a> Protein Cell. 6 (2), 88-100 (2015).</li><li>Torti, S. V., Manz, D. H., Paul, B. T., Blanchette-Farra, N., Torti, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+and+cancer.">Iron and cancer.</a> Annu Rev Nutr. 38, 97-125 (2018).</li><li>Welch, D. R., Hurst, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+the+hallmarks+of+metastasis.">Defining the hallmarks of metastasis.</a> Cancer Res. 79 (12), 3011-3027 (2019).</li><li>Lehmann, U., 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=Epigenetic+defects+of+hepatocellular+carcinoma+are+already+found+in+non-neoplastic+liver+cells+from+patients+with+hereditary+haemochromatosis.">Epigenetic defects of hepatocellular carcinoma are already found in non-neoplastic liver cells from patients with hereditary haemochromatosis.</a> Hum Mol Genet. 16 (11), 1335-1342 (2007).</li><li>Fonseca-Nunes, A., Jakszyn, P., Agudo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+and+cancer+risk--a+systematic+review+and+meta-analysis+of+the+epidemiological+evidence.">Iron and cancer risk--a systematic review and meta-analysis of the epidemiological evidence.</a> Cancer Epidemiol Biomarkers Prev. 23 (1), 12-31 (2014).</li><li>Abdelaal, G., Veuger, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversing+oncogenic+transformation+with+iron+chelation.">Reversing oncogenic transformation with iron chelation.</a> Oncotarget. 12 (2), 106-124 (2021).</li><li>Brown, R. A. 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=Altered+iron+metabolism+and+impact+in+cancer+biology,+metastasis,+and+immunology.">Altered iron metabolism and impact in cancer biology, metastasis, and immunology.</a> Front Oncol. 10, 476 (2020).</li><li>Menyh&#225;rt, O., 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=Guidelines+for+the+selection+of+functional+assays+to+evaluate+the+hallmarks+of+cancer.">Guidelines for the selection of functional assays to evaluate the hallmarks of cancer.</a> Biochim Biophys Acta. 1866 (2), 300-319 (2016).</li><li>Basu, 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=Discovering+novel+and+diverse+iron-chelators+in+silico.">Discovering novel and diverse iron-chelators in silico.</a> J Chem Inf Model. 56 (12), 2476-2485 (2016).</li><li>Prus, E., Fibach, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+cytometry+measurement+of+the+labile+iron+pool+in+human+hematopoietic+cells.">Flow cytometry measurement of the labile iron pool in human hematopoietic cells.</a> Cytometry A. 73 (1), 22-27 (2008).</li><li>Cold Spring Harb. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Phosphate-buffered saline (pbs). 2006 (1), (2006).</li><li>Kontoghiorghes, G. J., Pattichis, K., Neocleous, K., Kolnagou, 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+design+and+development+of+deferiprone+(l1)+and+other+iron+chelators+for+clinical+use:+Targeting+methods+and+application+prospects.">The design and development of deferiprone (l1) and other iron chelators for clinical use: Targeting methods and application prospects.</a> Curr Med Chem. 11 (16), 2161-2183 (2004).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Ammonium iron(II) sulfate hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>215406</td>
			<td>other wise known as FAS</td>
		</tr>
		<tr>
			<td>Calcein</td>
			<td>Sigma-Aldrich</td>
			<td>C0875</td>
			<td></td>
		</tr>
		<tr>
			<td>Deferiprone</td>
			<td>Sigma-Aldrich</td>
			<td>379409</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#8242;s Phosphate Buffered Saline</td>
			<td>Sigma-Aldrich</td>
			<td>D5652</td>
			<td>magnesium and calcium free</td>
		</tr>
		<tr>
			<td>Greiner CELLSTAR 96 well plates</td>
			<td>Sigma-Aldrich</td>
			<td>M0812</td>
			<td>any optically transparent 96 well plate will work</td>
		</tr>
	</tbody>
</table>

quantifiable and inexpensive cell-free fluorescent method to confirm the ability of novel compounds to chelate iron 

Phenotypic changes to cells that relate to the development of cancer through a common set of altered biological capabilities are now commonly referred to as the hallmarks of cancer. Amongst them are changes resulting from the reprogramming of energy metabolism, which are widespread in cancer cell biology1. Such metabolic reprogramming includes an increased requirement for iron to support rapid proliferation and tumor growth2. This thirst for iron leads to dysregulated iron metabolism, which in and of itself is considered a hallmark of cancer3,4, with dysregulation occurring at all stages5. The hallmarks of metastasis, more recently proposed by Welch and Hurst, include a role for iron6 since iron can induce oxidative stress, and this can, in turn, mediate changes to the genome, epigenome, and proteome, enhancing the possibility of metastasis7. A link between iron levels and an increased occurrence of cancer has been demonstrated through epidemiological studies8.
Since cancer cells require large amounts of iron, they are susceptible to iron deficiency and, therefore, iron chelation. We have recently published a review article highlighting the potential for iron chelation in reversing several hallmarks of cancer through NDRG1 disrupting oncogenic signaling pathways9. However, the use of iron chelation as stand-alone cancer therapy has not yielded positive results in clinical trials due to their toxicity, short half-life, rapid metabolism, and emerging resistance mechanisms. Nevertheless, iron chelators have shown promise in in vitro and in vivo investigations, indicating that more work is needed to develop effective iron chelators for cancer therapy. Specific iron chelation is a validated strategy in anticancer drug discovery, but only a few classes have been reported to date10.
The identification and characterization of novel iron chelators requires the ability to measure their effect on several endpoints. Many of these (such as proliferation, apoptosis, reactive oxygen species formation) are routinely measured and have been outlined and reviewed in the literature as methods to evaluate the hallmarks of cancer11. When evaluating a novel iron chelator, many groups routinely examine the effect on anti-proliferative and redox activities as well as effects on iron influx or efflux. In silico prediction techniques12 further increase the growing pool of iron chelators that can be screened.
Screening of iron chelators requires the ability to measure their effect on iron levels as a means to effectively demonstrate proof of principle. Currently, the most common method to do so is flow cytometry13 which is costly, time consuming and poorly quantifiable. The choice of assay is often based on the availability of experimental equipment, speed, and cost of an assay. Therefore, the ability to chelate iron can be an overlooked end point measure due to the high costs of equipment and the challenge to quickly and reproducibly quantify the strength of chelation. Here, we describe a quantifiable and inexpensive cell free fluorescent method to confirm the ability of novel compounds to chelate iron.

<meta charset="utf8"></head><body>Yazar Spotlight: Yeni Demir Şelatörlerinin Kanser Hücresi Metabolizması Üzerindeki Etkisinin Değerlendirilmesi

Yeni bileşiklerin demiri şelatlama yeteneğini doğrulamak için ölçülebilir ve ucuz hücresiz floresan yöntemi 

We developed a fluorescent assay that can quickly and inexpensively confirm the ability of novel compounds to chelate iron. Iron chelators are a promising class of compound that can be used to target cancers. Chelators'disruption of cancer's metabolic adaptions may lead to a new therapeutic strategy in cancer treatment.There is a research focus on the development of iron chelators to disrupt the metabolic adaptions of cancers. However, there are no inexpensive and simple screening techniques for the observation of a novel molecule's ability to chelate iron. There is an unmet need for a simple, inexpensive assay for screening novel iron chelators.This assay will allow cancer biologists to observe the chelation ability of novel compounds before investing time and resource into cellular screens of biological activity. Using this assay, we were able to observe the iron chelation ability of newly developed iron chelators. We have then focused on the question of how these chelators disrupt metabolism in cancer cells and how these cells respond to this disruption.We are fascinated with how cancer cells respond to the metabolic disruptions created by the treatment with iron chelators. Understanding iron metabolism in cancers is fundamental to the development of novel therapeutics and new treatment regimes.

Read this Scientific Article Protocol about Quantifiable and Inexpensive Cell-Free Fluorescent Method to Confirm the Ability of Novel Compounds to Chelate Iron at JoVE.com

Cancer cells require large amounts of iron to maintain their proliferation. Iron metabolism is considered a hallmark of cancer, making iron a valid target ...

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Research

JoVE Journal

Cancer Research

Quantifiable and Inexpensive Cell-Free Fluorescent Method to Confirm the Ability of Novel Compounds to Chelate Iron 

237 Views.  Northumbria University. We developed a fluorescent assay that can quickly and inexpensively confirm the ability of novel compounds to chelate iron. Iron chelators are a promising class of compound that can be used to target cancers. Chelators'disruption of cancer's metabolic adaptions may lead to a new therapeutic strategy in cancer treatment.There is a research focus on the development of iron chelators to disrupt the metabolic adaptions of cancers. However, there are no inexpensive and simple screening tech...