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Naoghare, Pravin K.,Tak, Yu Kyung,Kim, Min Jung,Han, Eunyoung,Song, Joon Myong Blackwell Publishing Ltd 2011 Basic & Clinical Pharmacology & Toxicology Vol.109 No.4
<P><B>Abstract: </B> Understanding the role of oncomirs allows new insights into the development of modern therapeutic approaches for the repression of multiple oncomirs in cancer cells. At present, no suitable approach is available to repress the development of multiple oncomirs in cancer cells. Herein, we report that argonaute 2 (AGO2) could be a unique molecule to regulate the development of multiple oncomirs in cancer cells. Knock‐down of AGO2 by custom‐made AGO2 siRNA resulted in the induction of apoptosis in myeloid leukaemia cells (HL‐60). Further investigations revealed that knock‐down of AGO2 by custom‐made AGO2 siRNA in HEK‐293 cells resulted in silencing of the expression of target genes vascular endothelial growth factor A and histone deacetylase 2, which are known to be involved in the development of myeloid leukaemia. From these results, it can be predicted that AGO2 could regulate siRNA‐mediated RNAi pathways in cancer cells. Furthermore, we investigated the possible implication of AGO2 in drug‐induced apoptosis. Investigations revealed that treatment with the newly synthesized drug analogue SH‐03[{(7S,7aR,13aS)‐9,10‐dimethoxy‐3,3‐dimethyl‐7,7a,13,13atetrahydro‐3H‐chromeno[3,4‐b]pyrano[2,3‐h]chromen‐7‐ol}] could induce AGO2‐mediated apoptosis in myeloid leukaemia cells via intrinsic apoptotic pathways independent of Dicer.</P>
Simultaneous quantitative monitoring of drug-induced caspase cascade pathways in carcinoma cells
Naoghare, Pravin K.,Ki, Hyeon A.,Paek, Seung-Mann,Tak, Yu Kyung,Suh, Young-Ger,Kim, Sang Geon,Lee, Kyeong-Hee,Song, Joon Myong Royal Society of Chemistry 2010 Integrative biology Vol.2 No.1
<P>Caspases are the key mediators of apoptosis. The caspase cascade includes a series of events leading to the activation of initiator and downstream caspases in a cell. Analysis of the caspase cascade in intact cells, however, has generally been limited as the simultaneous monitoring of upstream and downstream caspases is not well executed. In an effort to monitor the activation of caspase cascades in an intact cell, high-content cellular imaging that allows simultaneous quantitative monitoring of caspase activation has been developed. This has great significance for the exploration of various cellular caspases involved in apoptotic pathways as possible therapeutic targets in the process of drug discovery. To explore the potential of simultaneous monitoring of caspase-mediated apoptotic pathways, human myeloid leukemia HL-60 cells were treated with SH-03 {(7S,7<I>aR</I>,13<I>aS</I>)-9,10-dimethoxy-3,3-dimethyl-7,7<I>a</I>,13,13<I>a</I>-tetrahydro-3<I>H</I>-chromeno [3,4-<I>b</I>]pyrano[2,3-<I>h</I>]chromen-7-ol} (a newly synthesized candidate), camptothecin or naringenin (agents known to induce apoptosis) with or without caspase inhibitors. SH-03 or naringenin treatment initiated the caspase cascade through an intrinsic apoptotic pathway, whereas camptothecin treatment triggered both intrinsic and extrinsic caspase cascades. We now report a new approach based on uniform threshold intensity distribution that facilitates rapid, quantitative monitoring of drug-induced caspase cascades through multi-spectral and multicolor imaging cytometry.</P> <P>Graphic Abstract</P><P>Simultaneous monitoring of caspase cascades. The developed concept may have fundamental significance in establishing image-based assays for elucidating simultaneous cellular events. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=b916481b'> </P>