Grape Proanthocyanidin Inhibit Pancreatic Cancer Cell Growth In Vitro and In Vivo through Induction of Apoptosis and by Targeting the PI3K/Akt Pathway
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{"title"=>"Grape proanthocyanidin inhibit pancreatic cancer cell growth in vitro and in vivo through induction of apoptosis and by targeting the PI3K/Akt pathway", "type"=>"journal", "authors"=>[{"first_name"=>"Ram", "last_name"=>"Prasad", "scopus_author_id"=>"48661462100"}, {"first_name"=>"Mudit", "last_name"=>"Vaid", "scopus_author_id"=>"15837951100"}, {"first_name"=>"Santosh K.", "last_name"=>"Katiyar", "scopus_author_id"=>"7102906169"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"365400242", "doi"=>"10.1371/journal.pone.0043064", "sgr"=>"84864709263", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"22905202", "scopus"=>"2-s2.0-84864709263"}, "id"=>"af5f2e06-f7fb-3203-ac96-5652ff971683", "abstract"=>"Pancreatic cancer is an aggressive malignancy that is frequently diagnosed at an advanced stage with poor prognosis. Here, we report the chemotherapeutic effects of bioactive proanthocyanidins from grape seeds (GSPs) as assessed using In Vitro and In Vivo models. Treatment of human pancreatic cancer cells (Miapaca-2, PANC-1 and AsPC-1) with GSPs In Vitro reduced cell viability and increased G2/M phase arrest of the cell cycle leading to induction of apoptosis in a dose- and time-dependent manner. The GSPs-induced apoptosis of pancreatic cancer cells was associated with a decrease in the levels of Bcl-2 and Bcl-xl and an increase in the levels of Bax and activated caspase-3. Treatment of Miapaca-2 and PANC-1 cells with GSPs also decreased the levels of phosphatidylinositol-3-kinase (PI3K) and phosphorylation of Akt at ser(473). siRNA knockdown of PI3K from pancreatic cancer cells also reduced the phosphorylation of Akt. Further, dietary administration of GSPs (0.5%, w/w) as a supplemented AIN76A control diet significantly inhibited the growth of Miapaca-2 pancreatic tumor xenografts grown subcutaneously in athymic nude mice, which was associated with: (i) inhibition of cell proliferation, (ii) induction of apoptosis of tumor cells, (iii) increased expression of Bax, reduced expression of anti-apoptotic proteins and activation of caspase-3-positive cells, and (iv) decreased expression of PI3K and p-Akt in tumor xenograft tissues. Together, these results suggest that GSPs may have a potential chemotherapeutic effect on pancreatic cancer cell growth.", "link"=>"http://www.mendeley.com/research/grape-proanthocyanidin-inhibit-pancreatic-cancer-cell-growth-vitro-vivo-through-induction-apoptosis-4", "reader_count"=>21, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Ph. D. Student"=>5, "Student > Master"=>5, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Ph. D. Student"=>5, "Student > Master"=>5, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>10, "Medicine and Dentistry"=>5, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Taiwan"=>1}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/594743"], "description"=>"<p>(<b>A</b>) Miapaca-2 and PANC-1 cells were treated with different concentrations of GSPs (0, 20, 40 and 60 µg/mL) in complete medium for 48 h, then harvested for the analysis of apoptotic cells by FACS analysis using the Annexin V-Alexa Fluor488 Apoptosis Vybrant Assay Kit (Alexa488) following the manufacturer's protocol. The lower right (LR) quadrant of the FACS histograms indicates the percentage of early apoptotic cells (Alexa488-stained cells) and the upper right (UR) quadrant indicates the percentage of late apoptotic cells (Alexa488+ propidium iodide-stained cells). (<b>B</b>) Total percentages of apoptotic cells in each treatment group after 48 h are summarized with data presented as the mean ± SD of two experiments. Significant difference <i>vs.</i> control group (non-GSPs-treated), *<i>P</i><0.001; **<i>P</i><0.05. (<b>C</b>) Treatment of Miapaca-2 and PANC-1 cells with varying concentrations of GSPs for 48 h results in a dose-dependent reduction in the expression levels of the anti-apoptotic proteins (Bcl-2 and Bcl-xl) while increasing the expression of the pro-apoptotic protein Bax. GSPs also increase the activation level of caspase-3 in cells.</p>", "links"=>[], "tags"=>["induce", "apoptosis", "pancreatic", "cancer", "cells", "dose-dependent"], "article_id"=>265232, "categories"=>["Cancer", "Cell Biology"], "users"=>["Ram Prasad", "Mudit Vaid", "Santosh K. Katiyar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0043064.g003", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GSPs_induce_apoptosis_in_human_pancreatic_cancer_cells_in_a_dose_dependent_manner_/265232", "title"=>"GSPs induce apoptosis in human pancreatic cancer cells in a dose-dependent manner.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 01:07:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/594835"], "description"=>"<p>(<b>A</b>) Treatment of Miapaca-2 and PANC-1 cells with GSPs decreases the expression level of PI3Kp85 (regulatory) and PI3Kp110 (catalytic), and phosphorylation of Akt in a dose-dependent manner. After treatment for 48 h, cells were harvested, and cell lysates were subjected to western blot analysis to determine the expression of different proteins. β-actin was used to verify equal loading of the protein samples. (<b>B</b>) siRNA knockdown of PI3K in pancreatic cancer cells resulted in reduction of p-Akt levels compared to siRNA control group.</p>", "links"=>[], "tags"=>["gsps", "PI3K", "signaling", "proteins", "pancreatic", "cancer"], "article_id"=>265328, "categories"=>["Cancer", "Cell Biology"], "users"=>["Ram Prasad", "Mudit Vaid", "Santosh K. Katiyar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0043064.g004", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_GSPs_on_PI3K_signaling_proteins_in_pancreatic_cancer_cells_/265328", "title"=>"Effect of GSPs on PI3K signaling proteins in pancreatic cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 01:07:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/594931"], "description"=>"<p>(<b>A</b>) Dietary administration of GSPs (0.5%, w/w) inhibits the growth of Miapaca-2 cells grown as xenografts in athymic nude mice. Average tumor volume ± SD/mouse (mm3) in each treatment group was measured on weekly basis. Tumor xenograft tissues were harvested at the termination of the experiment at 11 weeks and the wet weight of the tumor/mouse in each group is reported in grams as the mean ± SD. Statistical significance of difference between control and GSPs-treated groups was analyzed by one-way ANOVA followed by the Bonferroni <i>t</i> test, n = 8. Statistical significance <i>vs</i> tumors from non-GSPs-treated control mice, *<i>P</i><0.005. (<b>B</b>) The immunohistochemical detection of TUNEL-positive cells in tumor xenograft tissues from mice administered GSPs in the diet and mice not receiving GSPs are shown, and resultant data on TUNEL-positive cells are summarized. (<b>C</b>) Immunohistochemical detection of activated caspase-3-positive cells in tumor xenograft tissues from GSPs-treated and non-GSPs-treated mice. Immunohistochemical data in terms of percentage of positive cells are presented as the mean ± SD of 5–6 tumor samples from each group. TUNEL-positive or caspase-3-positive cells were counted in four to five different fields, and data are summarized in terms of percent positive cells from 5–6 tumor xenograft samples. Representative micrographs are shown. (<b>D</b>) Dietary GSPs inhibit the expression of anti-apoptotic proteins while increasing the expression of pro-apoptotic protein, Bax, and the levels of activated caspases-3 in tumor xenograft tissues. Tumor lysates were prepared from the tumor xenografts collected at the termination of the experiment described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043064#pone-0043064-g005\" target=\"_blank\">Figure 5A</a> and subjected to western blot analysis, as described in Materials and methods. Representative blots are shown from independent experiments from at least five tumors from five different mice per group with identical observations.</p>", "links"=>[], "tags"=>["gsps", "inhibit", "xenograft", "athymic", "nude"], "article_id"=>265428, "categories"=>["Cancer", "Cell Biology"], "users"=>["Ram Prasad", "Mudit Vaid", "Santosh K. Katiyar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0043064.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dietary_GSPs_inhibit_in_vivo_tumor_xenograft_growth_in_athymic_nude_mice_/265428", "title"=>"Dietary GSPs inhibit <i>in vivo</i> tumor xenograft growth in athymic nude mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 01:08:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/595036"], "description"=>"<p>(<b>A</b>) Immunohistochemical analysis of tumor samples obtained from mice at the termination of the experiment described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043064#pone-0043064-g005\" target=\"_blank\">Figure 5</a> was used to detect Ki-67-positive cells as an estimate of the proliferation index. Ki-67-positive cells were counted in four to five different fields of a section, and data are summarized in terms of percent positive cells from 5 tumor xenograft samples. (<b>B</b>) Dietary GSPs reduce the levels of PI3K (p110, p85) and p-Akt in tumor xenograft tissues. Western blot analysis was performed to analyze the levels of these proteins. (<b>C</b>) Dietary GSPs decrease the levels of p-Akt in tumor samples. Immunohistochemical data show a reduction in the immunostaining intensity of p-Akt in tumor xenograft samples from GSPs-treated mice. Representative micrographs are shown from 4 tumor samples/group. Phospho-Akt-positive cells were counted in four to five different fields of a section, and data are summarized in terms of percent positive cells from 5 tumor xenograft samples (right panel). Statistical significance <i>vs</i>. tumor samples from mice receiving the control diet only, <sup>*</sup><i>P</i><0.001.</p>", "links"=>[], "tags"=>["gsps", "proliferation", "inhibit", "PI3K", "signaling", "pathway", "xenograft"], "article_id"=>265526, "categories"=>["Cancer", "Cell Biology"], "users"=>["Ram Prasad", "Mudit Vaid", "Santosh K. Katiyar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0043064.g006", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dietary_GSPs_reduce_the_proliferation_potential_and_inhibit_PI3K_signaling_pathway_in_xenograft_tissues_/265526", "title"=>"Dietary GSPs reduce the proliferation potential and inhibit PI3K signaling pathway in xenograft tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 01:08:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/594655"], "description"=>"<p>Miapaca-2 and PANC-1 cells were treated either with vehicle (0.1% DMSO) or GSPs (20, 40 and 60 µg/mL) in complete medium. After 48 h of treatment, cells were harvested and digested with RNase. Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution, as detailed in the Materials and Methods. The representative cell cycle histograms from two independent experiments in Miapaca-2 (<b>A</b>) and PANC-1 (<b>B</b>) cells after treatment with different doses of GSPs are shown. (<b>C</b>) Effect of GSPs on G2/M phase cell cycle regulatory proteins in Miapaca-2 and PANC-1 cells. The cells were treated with either vehicle or GSPs (20, 40, 60 µg/mL in DMSO) for 48 h and thereafter harvested, cell lysates prepared and then subjected to SDS-PAGE followed by western blot analyses for various proteins. β-actin was used to verify equal loading of the samples. Representative blots from three individual experiments are shown.</p>", "links"=>[], "tags"=>["gsps", "progression", "pancreatic", "cancer"], "article_id"=>265147, "categories"=>["Cancer", "Cell Biology"], "users"=>["Ram Prasad", "Mudit Vaid", "Santosh K. Katiyar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0043064.g002", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_GSPs_on_cell_cycle_progression_of_pancreatic_cancer_cells_/265147", "title"=>"Effect of GSPs on cell cycle progression of pancreatic cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 01:06:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/594564"], "description"=>"<p>(<b>A</b>) Miapaca-2 cells, (<b>B</b>) PANC-1 cells, and (<b>C</b>) AsPC-1 cells were treated with the indicated concentrations of GSPs for the indicated time periods. Left panels: Viability of the cells was determined using the MTT assay as described in the Materials and Methods. The data are expressed in terms of percent of control cells (non-GSPs treated) as the mean ± SD of 8 replicates. Right panels: The cytotoxic effect of GSPs on pancreatic cancer cells was determined using the trypan blue dye exclusion assay as described in Materials and methods. The cell death data are presented as the mean percent of dead cells from three experiments ± SD <i>vs.</i> control group (non-GSPs-treated). Significant difference <i>vs.</i> control group, <sup>*</sup><i>P</i><0.001; <sup>**</sup><i>P</i><0.05.</p>", "links"=>[], "tags"=>["pancreatic", "cancer", "cells", "gsps", "proliferation", "induces"], "article_id"=>265052, "categories"=>["Cancer", "Cell Biology"], "users"=>["Ram Prasad", "Mudit Vaid", "Santosh K. Katiyar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0043064.g001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Treatment_of_pancreatic_cancer_cells_with_GSPs_reduces_their_proliferation_potential_and_induces_cell_death_/265052", "title"=>"Treatment of pancreatic cancer cells with GSPs reduces their proliferation potential and induces cell death.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 01:05:58"}

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Relative Metric

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