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KYSE450 were purchased from Die Deutsche Managementsystem Zertifizierungsgesellschaft mbH (DSMZ, Braunschweig, Germany)." ; ns1:mentions . _:b6156047 rdf:type ns4:Section ; dc:title "results" ; ns4:contains _:b6156056 , _:b6156057 , _:b6156058 , _:b6156059 , _:b6156060 , _:b6156061 , _:b6156062 , _:b6156048 , _:b6156049 , _:b6156050 , _:b6156051 , _:b6156052 , _:b6156053 , _:b6156054 , _:b6156055 . _:b6156048 rdf:type ns1:Context ; rdf:value "MPM-2 is an antibody that recognises a group of proteins whose epitopes are exclusively phosphorylated during mitosis \u2013 particularly early prophase to metaphase (Davis et al, >>1983<<) and it is commonly used as an indicator of mitotic disturbance." ; ns1:mentions . _:b6156049 rdf:type ns1:Context ; rdf:value "As a positive control, cells were treated with nocodazole, a known inducer of metaphase arrest (Vassilev et al, >>2006<<). Treatment of KYSE450 cells with nocodazole for 18\u2009h resulted in a synchronisation of the entire cell population in the G2/M phase and an increase in MPM-2 labelling from 0.15 to 23.5% (Figure 2B). In all cells treated with 25\u2009\u03BCM of" ; ns1:mentions . _:b6156050 rdf:type ns1:Context ; rdf:value "Other studies have reported that curcumin can induce MC, which is currently defined primarily by morphology (Kroemer et al, >>2009<<). We therefore examined the morphology of curcumin-treated cells. Figure 3A shows representative examples of both treated and untreated cell lines. After 24\u2009h treatment with concentrations as low as 15\u2009\u03BCM, a marked elevation in the number" ; ns1:mentions . _:b6156051 rdf:type ns1:Context ; rdf:value "The dramatic vacuolisation of the cytoplasm without apparent loss of nuclear material is consistent with the described macrostructure of autophagy (F) (Figure 3A) (Kroemer et al, >>2009<<). This type of autophagic morphology in the KYSE450 cell line has also been observed by our group in response to treatment with cytotoxic drugs such as 5-fluorouracil and cisplatin (unpublished data). Interestingly, the features of MC and" ; ns1:mentions . _:b6156052 rdf:type ns1:Context ; rdf:value "Various papers have suggested that MC is an independent death process, whereas others have proposed that MC is just a consequence of mitotic failure that leads to apoptosis (Castedo et al, >>2004<<; Vakifahmetoglu et al, 2008). We therefore looked for the presence of active caspase-3, which is an indicator of classical apoptosis." ; ns1:mentions . _:b6156053 rdf:type ns1:Context ; rdf:value "Various papers have suggested that MC is an independent death process, whereas others have proposed that MC is just a consequence of mitotic failure that leads to apoptosis (Castedo et al, 2004; Vakifahmetoglu et al, >>2008<<). We therefore looked for the presence of active caspase-3, which is an indicator of classical apoptosis." ; ns1:mentions . _:b6156054 rdf:type ns1:Context ; rdf:value "Autophagy is a process that sequesters cytoplasmic proteins or organelles into a lytic compartment that facilitates degradation and re-cycling (Xie and Klionsky, >>2007<<). This can promote cell survival during cell stress; however, excessive autophagy can also lead to cell death (Degterev and Yuan, 2008). To investigate whether autophagy coexisted in the cell lines after curcumin treatment, we used the" ; ns1:mentions . _:b6156055 rdf:type ns1:Context ; rdf:value "This can promote cell survival during cell stress; however, excessive autophagy can also lead to cell death (Degterev and Yuan, >>2008<<). To investigate whether autophagy coexisted in the cell lines after curcumin treatment, we used the MDC assay, which is a selective fluorescent marker of autophagic vesicles (Biederbick et al, 1995). The dye shows diffuse staining in" ; ns1:mentions . _:b6156056 rdf:type ns1:Context ; rdf:value "To investigate whether autophagy coexisted in the cell lines after curcumin treatment, we used the MDC assay, which is a selective fluorescent marker of autophagic vesicles (Biederbick et al, >>1995<<). The dye shows diffuse staining in non-autophagic cells, but is sequestered into punctate vesicular staining when autophagy is induced. We therefore examined distribution of the dye in the three curcumin-sensitive cell lines (OE21, OE33" ; ns1:mentions . _:b6156057 rdf:type ns1:Context ; rdf:value "The primary target of curcumin is unknown; however, an emerging body of evidence suggests that this compound may achieve its diverse effects through inhibition of the UPS (Henke et al, >>1999<<; Uhle et al, 2003; Jana et al, 2004; Si et al, 2007)." ; ns1:mentions . _:b6156058 rdf:type ns1:Context ; rdf:value "The primary target of curcumin is unknown; however, an emerging body of evidence suggests that this compound may achieve its diverse effects through inhibition of the UPS (Henke et al, 1999; Uhle et al, >>2003<<; Jana et al, 2004; Si et al, 2007)." ; ns1:mentions . _:b6156059 rdf:type ns1:Context ; rdf:value "The primary target of curcumin is unknown; however, an emerging body of evidence suggests that this compound may achieve its diverse effects through inhibition of the UPS (Henke et al, 1999; Uhle et al, 2003; Jana et al, >>2004<<; Si et al, 2007)." ; ns1:mentions . _:b6156060 rdf:type ns1:Context ; rdf:value "primary target of curcumin is unknown; however, an emerging body of evidence suggests that this compound may achieve its diverse effects through inhibition of the UPS (Henke et al, 1999; Uhle et al, 2003; Jana et al, 2004; Si et al, >>2007<<). We therefore looked for evidence of the accumulation of poly-ubiquitinated proteins after treatment." ; ns1:mentions . _:b6156061 rdf:type ns1:Context ; rdf:value "UPS-mediated degradation of cyclin B1 is then required for cell-cycle progression through mitosis (Kops et al, >>2005<<; Vassilev et al, 2006)." ; ns1:mentions . _:b6156062 rdf:type ns1:Context ; rdf:value "UPS-mediated degradation of cyclin B1 is then required for cell-cycle progression through mitosis (Kops et al, 2005; Vassilev et al, >>2006<<). If the UPS system is inhibited in these cell lines, then cyclin B1 is likely to accumulate and may contribute to the major disturbances in mitotic checkpoints. We therefore examined expression after treatment with 15 and 25\u2009\u03BCM of" ; ns1:mentions . _:b6156063 rdf:type ns4:Section ; dc:title "discussion" ; ns4:contains _:b6156088 , _:b6156089 , _:b6156090 , _:b6156091 , _:b6156092 , _:b6156093 , _:b6156094 , _:b6156095 , _:b6156080 , _:b6156081 , _:b6156082 , _:b6156083 , _:b6156084 , _:b6156085 , _:b6156086 , _:b6156087 , _:b6156072 , _:b6156073 , _:b6156074 , _:b6156075 , _:b6156076 , _:b6156077 , _:b6156078 , _:b6156079 , _:b6156064 , _:b6156065 , _:b6156066 , _:b6156067 , _:b6156068 , _:b6156069 , _:b6156070 , _:b6156071 , _:b6156096 . _:b6156064 rdf:type ns1:Context ; rdf:value "Curcumin has shown potential as both a chemopreventive agent and a chemotherapeutic agent in various gastrointestinal cancers, and initial studies in oesophageal cancer have been encouraging (Ushida et al, >>2000<<). In this study, we have investigated the effects of curcumin on panel of genetically and phenotypically heterogeneous oesophageal cancer cell lines." ; ns1:mentions . _:b6156065 rdf:type ns1:Context ; rdf:value "The most sensitive cell lines (KYSE450, OE21 and OE33) underwent mitotic arrest with morphological features of MC (Castedo et al, >>2004<<; Vitale et al, 2005)." ; ns1:mentions . _:b6156066 rdf:type ns1:Context ; rdf:value "The most sensitive cell lines (KYSE450, OE21 and OE33) underwent mitotic arrest with morphological features of MC (Castedo et al, 2004; Vitale et al, >>2005<<). The published literature is divided as to whether MC is an event that precedes apoptotic cell death or is a distinct mechanism of cell death (reviewed in Vakifahmetoglu et al, 2008). Although morphological analysis did show apoptotic" ; ns1:mentions . _:b6156067 rdf:type ns1:Context ; rdf:value "The published literature is divided as to whether MC is an event that precedes apoptotic cell death or is a distinct mechanism of cell death (reviewed in Vakifahmetoglu et al, >>2008<<). Although morphological analysis did show apoptotic cells in OE21 and OE33, these are coexistent but not dominant features of curcumin-treated cell populations. If apoptosis was the only death mechanism, the low level of apoptosis is not" ; ns1:mentions . _:b6156068 rdf:type ns1:Context ; rdf:value "Other authors have reported biochemical features of apoptosis in cell populations showing MC morphology \u2013 primarily caspase activation, although indeed with minimal caspase 3 cleavage (Rashmi et al, >>2004<<, 2005). Another study also reported that caspase activation was not required for MC and that activation of caspases varied in different cell lines (Mansilla et al, 2006). Clearly, limited caspase activation in a heterogeneous population" ; ns1:mentions . _:b6156069 rdf:type ns1:Context ; rdf:value "Other authors have reported biochemical features of apoptosis in cell populations showing MC morphology \u2013 primarily caspase activation, although indeed with minimal caspase 3 cleavage (Rashmi et al, 2004, >>2005<<). Another study also reported that caspase activation was not required for MC and that activation of caspases varied in different cell lines (Mansilla et al, 2006). Clearly, limited caspase activation in a heterogeneous population of" ; ns1:mentions . _:b6156070 rdf:type ns1:Context ; rdf:value "Another study also reported that caspase activation was not required for MC and that activation of caspases varied in different cell lines (Mansilla et al, >>2006<<). Clearly, limited caspase activation in a heterogeneous population of cells (as in OE21 cells) does not necessarily mean that the MC cells are activating caspases. We have found minimal caspase 3 cleavage in two of our four cell lines" ; ns1:mentions . _:b6156071 rdf:type ns1:Context ; rdf:value "Other authors have recently reported exclusive autophagic cell death after curcumin treatment and this was associated with the activation of ERK signalling (Aoki et al, >>2007<<; Shinojima et al, 2007)." ; ns1:mentions . _:b6156072 rdf:type ns1:Context ; rdf:value "Other authors have recently reported exclusive autophagic cell death after curcumin treatment and this was associated with the activation of ERK signalling (Aoki et al, 2007; Shinojima et al, >>2007<<). However, we found de-phosphorylation of ERK after curcumin treatment (Supplementary data). It is possible that this difference is cell-line-related \u2013 or dose-related \u2013 as our study used much lower concentrations of drug (Aoki et al 80" ; ns1:mentions . _:b6156073 rdf:type ns1:Context ; rdf:value "A number of studies have also linked downregulation of survivin with MC (Magalska et al, >>2006<<; Wolanin et al, 2006)." ; ns1:mentions . _:b6156074 rdf:type ns1:Context ; rdf:value "A number of studies have also linked downregulation of survivin with MC (Magalska et al, 2006; Wolanin et al, >>2006<<). We did not see a correlation between low survivin expression and induction of MC in the oesophageal cancer cells (Supplementary data)." ; ns1:mentions . _:b6156075 rdf:type ns1:Context ; rdf:value "Our data are therefore supportive of the view that curcumin acts as an inhibitor of the UPS (Henke et al, >>1999<<; Jana et al, 2004; Si et al, 2007)." ; ns1:mentions . _:b6156076 rdf:type ns1:Context ; rdf:value "Our data are therefore supportive of the view that curcumin acts as an inhibitor of the UPS (Henke et al, 1999; Jana et al, >>2004<<; Si et al, 2007)." ; ns1:mentions . _:b6156077 rdf:type ns1:Context ; rdf:value "Our data are therefore supportive of the view that curcumin acts as an inhibitor of the UPS (Henke et al, 1999; Jana et al, 2004; Si et al, >>2007<<). This is likely to have marked effects on the cell cycle, but effects may differ in different cell lines because of differences in their existing proteome. For example, cells of a similar type that have lost transcription of tumour" ; ns1:mentions . _:b6156078 rdf:type ns1:Context ; rdf:value "Another reported activity of curcumin that may explain its pleiotropic nature is its effects on histone modification (Liu et al, >>2005<<). We cannot rule this out, and clearly more mechanistic work needs to be undertaken in this area. It is also noteworthy that the UPS system can influence chromatin structure and thus transcription (O\u2019Connell and Harper, 2007)." ; ns1:mentions . _:b6156079 rdf:type ns1:Context ; rdf:value "It is also noteworthy that the UPS system can influence chromatin structure and thus transcription (O\u2019Connell and Harper, >>2007<<)." ; ns1:mentions . _:b6156080 rdf:type ns1:Context ; rdf:value "At least three different Phase I clinical trials have indicated that curcumin is safe and well tolerated even at doses as high as 12\u2009g\u2009day\u22121 (Cheng et al, >>2001<<). However, despite its well-documented efficacy and safety in both animals and humans in vivo and in vitro, its poor bioavailability has been cited as a major concern and has probably delayed its development as a therapeutic agent. Serum" ; ns1:mentions . _:b6156081 rdf:type ns1:Context ; rdf:value "Serum concentrations of curcumin after oral administration range from 0.013 to 11.1\u2009\u03BCM (Anand et al, >>2007<<). Various studies have shown curcumin has low bioavailability, limited tissue distribution, short half-life and a rapid metabolism irrespective of its route of administration (Ravindranath and Chandrasekhara, 1981; Garcea et al, 2004," ; ns1:mentions . _:b6156082 rdf:type ns1:Context ; rdf:value "Various studies have shown curcumin has low bioavailability, limited tissue distribution, short half-life and a rapid metabolism irrespective of its route of administration (Ravindranath and Chandrasekhara, >>1981<<; Garcea et al, 2004, 2005; Hoehle et al, 2006; Anand et al, 2007)." ; ns1:mentions . _:b6156083 rdf:type ns1:Context ; rdf:value "Various studies have shown curcumin has low bioavailability, limited tissue distribution, short half-life and a rapid metabolism irrespective of its route of administration (Ravindranath and Chandrasekhara, 1981; Garcea et al, >>2004<<, 2005; Hoehle et al, 2006; Anand et al, 2007)." ; ns1:mentions . _:b6156084 rdf:type ns1:Context ; rdf:value "Various studies have shown curcumin has low bioavailability, limited tissue distribution, short half-life and a rapid metabolism irrespective of its route of administration (Ravindranath and Chandrasekhara, 1981; Garcea et al, 2004, >>2005<<; Hoehle et al, 2006; Anand et al, 2007). The fact that curcumin has poor systemic bioavailability would suggest that the gastrointestinal tract is the most promising site for use in prevention or therapy." ; ns1:mentions . _:b6156085 rdf:type ns1:Context ; rdf:value "shown curcumin has low bioavailability, limited tissue distribution, short half-life and a rapid metabolism irrespective of its route of administration (Ravindranath and Chandrasekhara, 1981; Garcea et al, 2004, 2005; Hoehle et al, >>2006<<; Anand et al, 2007). The fact that curcumin has poor systemic bioavailability would suggest that the gastrointestinal tract is the most promising site for use in prevention or therapy." ; ns1:mentions . _:b6156086 rdf:type ns1:Context ; rdf:value "low bioavailability, limited tissue distribution, short half-life and a rapid metabolism irrespective of its route of administration (Ravindranath and Chandrasekhara, 1981; Garcea et al, 2004, 2005; Hoehle et al, 2006; Anand et al, >>2007<<). The fact that curcumin has poor systemic bioavailability would suggest that the gastrointestinal tract is the most promising site for use in prevention or therapy." ; ns1:mentions . _:b6156087 rdf:type ns1:Context ; rdf:value "have gained attention, such as the use of piperine, nanoparticles, liposomes, phospholipid complexes and structurally altered derivatives and analogues that dramatically improve the biological activity of curcumin (Shoba et al, >>1998<<; Mishra et al, 2005; Liu et al, 2006; Anand et al, 2007; Li et al, 2007; Mosley et al, 2007)." ; ns1:mentions . _:b6156088 rdf:type ns1:Context ; rdf:value "such as the use of piperine, nanoparticles, liposomes, phospholipid complexes and structurally altered derivatives and analogues that dramatically improve the biological activity of curcumin (Shoba et al, 1998; Mishra et al, >>2005<<; Liu et al, 2006; Anand et al, 2007; Li et al, 2007; Mosley et al, 2007)." ; ns1:mentions . _:b6156089 rdf:type ns1:Context ; rdf:value "the use of piperine, nanoparticles, liposomes, phospholipid complexes and structurally altered derivatives and analogues that dramatically improve the biological activity of curcumin (Shoba et al, 1998; Mishra et al, 2005; Liu et al, >>2006<<; Anand et al, 2007; Li et al, 2007; Mosley et al, 2007)." ; ns1:mentions . _:b6156090 rdf:type ns1:Context ; rdf:value "nanoparticles, liposomes, phospholipid complexes and structurally altered derivatives and analogues that dramatically improve the biological activity of curcumin (Shoba et al, 1998; Mishra et al, 2005; Liu et al, 2006; Anand et al, >>2007<<; Li et al, 2007; Mosley et al, 2007)." ; ns1:mentions . _:b6156091 rdf:type ns1:Context ; rdf:value "liposomes, phospholipid complexes and structurally altered derivatives and analogues that dramatically improve the biological activity of curcumin (Shoba et al, 1998; Mishra et al, 2005; Liu et al, 2006; Anand et al, 2007; Li et al, >>2007<<; Mosley et al, 2007)." ; ns1:mentions . _:b6156092 rdf:type ns1:Context ; rdf:value "complexes and structurally altered derivatives and analogues that dramatically improve the biological activity of curcumin (Shoba et al, 1998; Mishra et al, 2005; Liu et al, 2006; Anand et al, 2007; Li et al, 2007; Mosley et al, >>2007<<). Combination studies so far in both in vitro and in vivo have highlighted promising combinations of curcumin with oxaliplatin (Howells et al, 2007; Li et al, 2007; Patel et al, 2008) and paclitaxel (Aggarwal et al, 2005), suggesting" ; ns1:mentions . _:b6156093 rdf:type ns1:Context ; rdf:value "Combination studies so far in both in vitro and in vivo have highlighted promising combinations of curcumin with oxaliplatin (Howells et al, >>2007<<; Li et al, 2007; Patel et al, 2008) and paclitaxel (Aggarwal et al, 2005), suggesting there is a very definite potential for curcumin to be used as part of mainstream clinical regimes." ; ns1:mentions . _:b6156094 rdf:type ns1:Context ; rdf:value "Combination studies so far in both in vitro and in vivo have highlighted promising combinations of curcumin with oxaliplatin (Howells et al, 2007; Li et al, >>2007<<; Patel et al, 2008) and paclitaxel (Aggarwal et al, 2005), suggesting there is a very definite potential for curcumin to be used as part of mainstream clinical regimes." ; ns1:mentions . _:b6156095 rdf:type ns1:Context ; rdf:value "Combination studies so far in both in vitro and in vivo have highlighted promising combinations of curcumin with oxaliplatin (Howells et al, 2007; Li et al, 2007; Patel et al, >>2008<<) and paclitaxel (Aggarwal et al, 2005), suggesting there is a very definite potential for curcumin to be used as part of mainstream clinical regimes." ; ns1:mentions . _:b6156096 rdf:type ns1:Context ; rdf:value "Combination studies so far in both in vitro and in vivo have highlighted promising combinations of curcumin with oxaliplatin (Howells et al, 2007; Li et al, 2007; Patel et al, 2008) and paclitaxel (Aggarwal et al, >>2005<<), suggesting there is a very definite potential for curcumin to be used as part of mainstream clinical regimes." ; ns1:mentions . _:b415462707 rdf:type ns1:RelevantBibliographicResource . @prefix xsd: . _:b415462707 ns1:RelevantScore "8"^^xsd:nonNegativeInteger ; ns1:hasRelevantPaperId . _:b415462708 rdf:type ns1:RelevantBibliographicResource ; 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