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PMC0
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10.1186%2F1757-2215-3-11
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materials and methods
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A2780 and A2780CP (resistant to cisplatin) paired cells [>>18<<] were generously provided by Dr. Stephen Howell, University of California, San Diego.
n2:mentions
n3:8106143
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Following treatment, cells were processed for protein extraction and Western blotting using standard procedures as described earlier [>>19<<]. Briefly, 800,000 cells per 100 mm cell culture dish were plated, allowed to attach overnight and treated with curcumin or cisplatin or pretreated with curcumin followed by cisplatin.
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n3:19176398
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The β-catenin-TCF transcription activity was measured using a luminescence reporter assay as described earlier [>>20<<]. In short, 200,000 cells were plated per well in a 12 well plate for 16 hrs prior to transient transfection with reporter construct TOPFlash or FOPFlash (Gift from Dr.
n2:mentions
n3:18765827
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nanoparticles (PLGA NPs) containing curcumin were prepared from curcumin and PLGA (50:50 lactide-glycolide ratio; inherent viscosity 1.32 dL/g in at 30°C) (Birmingham Polymers, Pelham, AL) using modified nano-precipitation technique [>>21<<]. In brief, 90 mg of PLGA was dissolved in 10 mL of acetone over a period of 3 hrs and 1 mg of curcumin was added to get a uniform PLGA-curcumin solution.
n2:mentions
n3:9971898
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Curcumin loading and release was estimated at 450 nm using Biomate 3 UV-vis spectrophotometer (Thermo Electron) as described earlier [>>22<<].
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The FITC loading in PLGA NPs was determined using UV-vis spectrophotometer [>>23<<] at 490 nm after extracting FITC for 1 day in acetone.
n2:mentions
n3:17949004
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results
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Inappropriate activation of β-catenin is linked with the development of a wide variety of cancers, including melanoma, colorectal and prostate cancer [>>24<<,25]. Additionally, deregulation of the Wnt/β-catenin pathway has also been shown in ovarian cancer [26,27].
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Inappropriate activation of β-catenin is linked with the development of a wide variety of cancers, including melanoma, colorectal and prostate cancer [24,>>25<<]. Additionally, deregulation of the Wnt/β-catenin pathway has also been shown in ovarian cancer [26,27].
n2:mentions
n3:14519384
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Additionally, deregulation of the Wnt/β-catenin pathway has also been shown in ovarian cancer [>>26<<,27]. As a modulator of the Wnt signaling pathway, β-catenin functions as a transcription factor that is translocated into the nucleus where it binds with the TCF transcription factor and up-regulates the expression of cell survival genes
n2:mentions
n3:17986238
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Additionally, deregulation of the Wnt/β-catenin pathway has also been shown in ovarian cancer [26,>>27<<]. As a modulator of the Wnt signaling pathway, β-catenin functions as a transcription factor that is translocated into the nucleus where it binds with the TCF transcription factor and up-regulates the expression of cell survival genes
n2:mentions
n3:16867867
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It has also been shown that β-catenin activity can also inhibit apoptosis in cancer cells [>>28<<-31]. Therefore, we sought to investigate the effects of curcumin treatment on nuclear β-catenin function in cisplatin resistant ovarian cancer cells using TOPFlash reporter assay. The cells were treated with either curcumin, cisplatin or
n2:mentions
n3:18487207 n3:16959035 n3:18680107 n3:16084063
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Therefore, we decided to synthesize a PLGA nanoparticle (NP) formulation of curcumin, which is expected to improve bioavailability in vivo [>>32<<,33]. Following synthesis, Nano-CUR was physically characterized by both dynamic light scattering (DLS) and transmission electron microscopy (TEM). The average size of Nano-CUR was observed to be ~72 nm by DLS (Figure 6A) and 70 ± 3.9 nm
n2:mentions
n3:19735646
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Therefore, we decided to synthesize a PLGA nanoparticle (NP) formulation of curcumin, which is expected to improve bioavailability in vivo [32,>>33<<]. Following synthesis, Nano-CUR was physically characterized by both dynamic light scattering (DLS) and transmission electron microscopy (TEM). The average size of Nano-CUR was observed to be ~72 nm by DLS (Figure 6A) and 70 ± 3.9 nm by
n2:mentions
n3:19491009
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TAG-72, a tumor-associated glycoprotein, is over-expressed in various tumors, including ovarian cancer [>>34<<]. Western blot analysis of conjugated PLGA NPs revealed that anti-TAG-72 MAb was effectively conjugated to PLGA NPs (Figure 6G). These data suggest that, in the future, targeted delivery of curcumin specifically to tumors will be possible.
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altered characteristics, including: (a) altered membrane transport properties, (b) altered expression of target enzymes, (c) promotion of DNA repair, (d) degradation of drug molecules, and (e) generalized resistance to apoptosis [>>35<<-37]. A promising strategy for improving current ovarian cancer therapy is to employ a chemo/radio-sensitizer along with chemo/radiation therapies.
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Curcumin is an excellent candidate as a chemo/radio sensitizer and has been shown to have in vitro chemo-sensitization effects for cervical cancer and radio-sensitizing effects for prostate cancer [>>38<<,39]. However, curcumin's utility for ovarian cancer treatment has not been fully explored [40-42].
n2:mentions
n3:14985701
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Curcumin is an excellent candidate as a chemo/radio sensitizer and has been shown to have in vitro chemo-sensitization effects for cervical cancer and radio-sensitizing effects for prostate cancer [38,>>39<<]. However, curcumin's utility for ovarian cancer treatment has not been fully explored [40-42].
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However, curcumin's utility for ovarian cancer treatment has not been fully explored [>>40<<-42]. Chirnomas et al. reported that a functional Fanconi anemia (FA)/BRCA pathway limits sensitivity to cisplatin and that curcumin can inhibit this pathway, leading to increased sensitivity to cisplatin treatment in ovarian cancer cells
n2:mentions
n3:19236381 n3:16648566 n3:12447990
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Chirnomas et al. reported that a functional Fanconi anemia (FA)/BRCA pathway limits sensitivity to cisplatin and that curcumin can inhibit this pathway, leading to increased sensitivity to cisplatin treatment in ovarian cancer cells [>>41<<]. Our study shows that a 6 hr pre-treatment with curcumin effectively sensitized cisplatin resistant ovarian cancer cells to the cytotoxic effects of cisplatin, at doses at least 10 times lower compared to cisplatin treatment alone.
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n3:16648566
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Over-expression of pro-survival Bcl2 family members is common in many types of cancer and has been correlated with decreased sensitivity to chemotherapy and radiation [>>43<<]. We found that curcumin pre-treatment reduced the expression of two pro-survival proteins, Bcl-XL and Mcl-1, potentially allowing curcumin treated cells to undergo apoptosis upon cisplatin treatment. Indeed, pre-treatment with curcumin
n2:mentions
n3:19156528
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The functions of these responsive genes are thought to increase proliferation and recent studies have also suggested that β-catenin signaling may also inhibit apoptosis [>>28<<-31]. Taken together, these results suggest that curcumin pre-treatment increases the effectiveness of cisplatin treatment in cisplatin resistant cells by increasing the sensitivity of cells to apoptotic pathways and modulating nuclear
n2:mentions
n3:18487207 n3:16959035 n3:18680107 n3:16084063
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Curcumin is in early phase clinical trials for various types of cancers [>>44<<]. Curcumin is remarkably well tolerated and has no toxicity issues [45,46], but it has limited bioavailability and poor pharamacokinetics [47,48].
n2:mentions
n3:18496811
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Curcumin is remarkably well tolerated and has no toxicity issues [>>45<<,46], but it has limited bioavailability and poor pharamacokinetics [47,48].
n2:mentions
n3:17569225
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_:vb7612723
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Curcumin is remarkably well tolerated and has no toxicity issues [45,>>46<<], but it has limited bioavailability and poor pharamacokinetics [47,48].
n2:mentions
n3:15501961
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Curcumin is remarkably well tolerated and has no toxicity issues [45,46], but it has limited bioavailability and poor pharamacokinetics [>>47<<,48]. To improve curcumin's in vivo effectiveness we have developed a PLGA nanoformulation of curcumin. Nanoparticles can deliver anti-cancer drugs to the site of disease with an antibody targeting approach; however, major drawbacks
n2:mentions
n3:11221833
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_:vb7612725
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Curcumin is remarkably well tolerated and has no toxicity issues [45,46], but it has limited bioavailability and poor pharamacokinetics [47,>>48<<]. To improve curcumin's in vivo effectiveness we have developed a PLGA nanoformulation of curcumin. Nanoparticles can deliver anti-cancer drugs to the site of disease with an antibody targeting approach; however, major drawbacks include
n2:mentions
n3:11815407
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to the site of disease with an antibody targeting approach; however, major drawbacks include interaction with serum proteins (causing opsonization), clearance by the reticuloendothelial system, and non specific accumulation in organs [>>49<<]. To counter these difficulties and to extend the circulation time of nanoparticles in the blood, nanoparticles may be modified with inert hydrophilic polymers, such as poly(ethylene glycol) and poly(vinyl alcohol).
n2:mentions
n3:19186176
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In addition, formulating a small particle size (less than 100 nm) with high antibody conjugation efficiency will further enhance the ability to target tumors efficiently [>>50<<]. In our current study, we have developed PLGA nanoparticles which are made using FDA approved polymer (PLGA) and coated with poly(vinyl alcohol). The formulated Nano-CUR effectively inhibits proliferation in cisplatin resistant ovarian
n2:mentions
n3:17454354
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Recent literature suggests that antibody conjugated nanoparticles could efficiently deliver chemotherapeutic drugs to the tumor site [>>51<<-53]. Accordingly, we have shown efficient conjugation of anti-TAG-72 MAb to PLGA NPs with our conjugation chemistry for targeting applications. Targeted delivery of curcumin will improve the therapeutic efficacy of curcumin and will be
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lethal gynecological cancer and the fifth most common cause of cancer mortality in women in the United States: in 2009 it is estimated that 21,550 women will be diagnosed with ovarian cancer and 14,600 women will die due to this disease [>>1<<]. A high percent of women with ovarian cancer are diagnosed at an advanced stage (67%) and have a 5 year survival rate of only 46% [1].
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A high percent of women with ovarian cancer are diagnosed at an advanced stage (67%) and have a 5 year survival rate of only 46% [>>1<<]. The usual treatment modality involves surgical cytoreduction followed by treatment with a combination of platinum (cisplatin or carboplatin) and taxane based therapies. This is effective in 60-80% of patients; however, the majority of
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This is effective in 60-80% of patients; however, the majority of women with advanced disease will have cancer recurrence [>>2<<,3]. Unfortunately, almost all relapsing ovarian cancers eventually develop platinum resistance and patients with resistant tumors have a median survival time of 6 months, with only 27% living longer than 12 months [4]. In addition to
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This is effective in 60-80% of patients; however, the majority of women with advanced disease will have cancer recurrence [2,>>3<<]. Unfortunately, almost all relapsing ovarian cancers eventually develop platinum resistance and patients with resistant tumors have a median survival time of 6 months, with only 27% living longer than 12 months [4]. In addition to
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n3:18416585
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Unfortunately, almost all relapsing ovarian cancers eventually develop platinum resistance and patients with resistant tumors have a median survival time of 6 months, with only 27% living longer than 12 months [>>4<<]. In addition to improving diagnosis of ovarian cancer, there is an urgent need to develop effective therapeutic modalities for advanced stage drug resistant ovarian cancer.
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Recently, it has been suggested that while initial sensitivity to cisplatin is via nonfunctional DNA repair genes (i.e. BRCA1/2), cisplatin resistance may be acquired through a gain of function in BRCA1/2 [>>5<<]. Independent of the mechanism of resistance, inhibition of cell death via apoptosis is an important event leading to cisplatin resistance. Another important aspect limiting the use of cisplatin is the negative side effects which
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aspect limiting the use of cisplatin is the negative side effects which accumulate in severity with multiple cisplatin treatments and include gastrointestinal distress, kidney and nerve damage, hearing loss, and bone marrow suppression [>>2<<,3,6]. Additionally, treatment of ovarian cancer with radiation is limited due to gastrointestinal toxicity [6].
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limiting the use of cisplatin is the negative side effects which accumulate in severity with multiple cisplatin treatments and include gastrointestinal distress, kidney and nerve damage, hearing loss, and bone marrow suppression [2,>>3<<,6]. Additionally, treatment of ovarian cancer with radiation is limited due to gastrointestinal toxicity [6].
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limiting the use of cisplatin is the negative side effects which accumulate in severity with multiple cisplatin treatments and include gastrointestinal distress, kidney and nerve damage, hearing loss, and bone marrow suppression [2,3,>>6<<]. Additionally, treatment of ovarian cancer with radiation is limited due to gastrointestinal toxicity [6].
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Additionally, treatment of ovarian cancer with radiation is limited due to gastrointestinal toxicity [>>6<<]. While significant progress has been made in developing targeted radioimmunotherapy (RIT), current drawbacks to this therapy include toxicity and resistance to radiation [7,8].
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While significant progress has been made in developing targeted radioimmunotherapy (RIT), current drawbacks to this therapy include toxicity and resistance to radiation [>>7<<,8].
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While significant progress has been made in developing targeted radioimmunotherapy (RIT), current drawbacks to this therapy include toxicity and resistance to radiation [7,>>8<<].
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A number of natural dietary phytochemicals, such as curcumin, quercetin, xanthorrhizol, ginger, green tea, genistein, etc., are candidates for inducing chemo/radio-sensitization of cancer cells [>>9<<-11]. Among these agents, curcumin (diferuloyl methane), a polyphenol derived from the rhizomes of tumeric, Curcuma longa, has received considerable attention due to its beneficial chemopreventive and chemotherapeutic activity via
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considerable attention due to its beneficial chemopreventive and chemotherapeutic activity via influencing multiple signaling pathways, including those involved in survival, growth, metastasis and angiogenesis in various cancers [>>12<<-15]. Importantly, curcumin has demonstrated no toxicity to healthy organs at doses as high as 8 grams/day [16].
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Importantly, curcumin has demonstrated no toxicity to healthy organs at doses as high as 8 grams/day [>>16<<]. However, the low bioavailability and poor pharmacokinetics of curcumin limits its effectiveness in vivo [17]; therefore, we have developed a PLGA nanoparticle formulation of curcumin (Nano-CUR) to provide increased bioavailability as
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However, the low bioavailability and poor pharmacokinetics of curcumin limits its effectiveness in vivo [>>17<<]; therefore, we have developed a PLGA nanoparticle formulation of curcumin (Nano-CUR) to provide increased bioavailability as well as antibody conjugation abilities for targeted delivery of curcumin into tumors.
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