Bacterial Quorum Sensing Molecule N-3-Oxo-Dodecanoyl-L-Homoserine Lactone Causes Direct Cytotoxicity and Reduced Cell Motility in Human Pancreatic Carcinoma Cells
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{"title"=>"Bacterial quorum sensing molecule N-3-oxo-dodecanoyl-L-homoserine lactone causes direct cytotoxicity and reduced cell motility in human pancreatic carcinoma cells", "type"=>"journal", "authors"=>[{"first_name"=>"Ashwath S.", "last_name"=>"Kumar", "scopus_author_id"=>"56351849400"}, {"first_name"=>"Jeffrey N.", "last_name"=>"Bryan", "scopus_author_id"=>"26660641400"}, {"first_name"=>"Senthil R.", "last_name"=>"Kumar", "scopus_author_id"=>"57195139583"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"600018043", "sgr"=>"84906964407", "issn"=>"19326203", "pmid"=>"25188245", "scopus"=>"2-s2.0-84906964407", "doi"=>"10.1371/journal.pone.0106480", "isbn"=>"10.1371/journal.pone.0106480"}, "id"=>"5d8bc4c7-bb8b-36e4-9124-5ea143724360", "abstract"=>"In spite of chemotherapeutic and surgical advances, pancreatic cancer continues to have a dismal prognosis. Metastasis due to tumor cell migration remains the most critical challenge in treating pancreatic cancer, and conventional chemotherapy is rarely curative. In the quest for more novel molecules to fight this disease, we tested the hypothesis that the Pseudomonas aeruginosa quorum sensing signal molecule N-3-oxo-dodecanoyl-L-homoserine lactone (O-DDHSL) would be cytotoxic to and reduce mobility of pancreatic carcinoma cells (Panc-1 and Aspc-1). Results showed a decrease in cell viability from apoptosis, diminished colony formation, and inhibition of migration of the evaluated pancreatic carcinoma cell lines. Also, cell viability decreased in the presence of O-DDHSL when cells were grown in matrigel basement membrane matrix. While messenger RNA for IQGAP-1 decreased in Panc-1 and HPDE cells upon exposure to O-DDHSL, no change was observed in Aspc-1 cells. Cofilin mRNA expression was found to be increased in both HPDE and Panc-1 cells with marginal decrease in Aspc-1 cells. RhoC, a Rho-family GTPase involved in cell motility, increased in the presence of O-DDHSL, suggesting a possible compensatory response to alteration in other migration associated genes. Our results indicate that O-DDHSL could be an effective biomolecule in eukaryotic systems with multimodal function for essential molecular targeting in pancreatic cancer.", "link"=>"http://www.mendeley.com/research/bacterial-quorum-sensing-molecule-n3oxododecanoyllhomoserine-lactone-causes-direct-cytotoxicity-redu", "reader_count"=>27, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Student > Doctoral Student"=>1, "Researcher"=>5, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>4, "Student > Bachelor"=>5}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Student > Doctoral Student"=>1, "Researcher"=>5, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>4, "Student > Bachelor"=>5}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>10, "Medicine and Dentistry"=>3, "Neuroscience"=>2, "Computer Science"=>1, "Immunology and Microbiology"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>4}}, "reader_count_by_country"=>{"United States"=>1, "South Africa"=>1, "Peru"=>1, "Russia"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1661555"], "description"=>"<p><b>A</b>- Panc-1, Aspc-1 and HPDE-1 cells (6×10<sup>3</sup> cells per well) in triplicates was treated with 300 (Panc-1 and HPDE) or 200 µM (Aspc-1) of O-DDHSL, respectively, for 24 h. Cell apoptosis was assayed using a fluorescent based caspase 3/7 activity detection kit. While apoptosis in both Panc-1 cells (P = 0.047*) and Aspc-1 cells (P≤0.02**) were significant, HPDE cell apoptosis was marginally significant (P = 0.059***) <b>B</b> – Light micrographs of Panc-1, Aspc-1 and HPDE cells before and after exposure to O-DDHSL at concentrations of 300 µM (Panc-1 and HPDE) and 200 µM (Aspc-1), respectively, for 48 h (20×). <b>C</b> – Cell proliferation of HPDE, Panc-1 and Aspc-1cells upon exposure to O-DDHSL decreased by 35% (P = 0.042*), 47% (P≤0.035 **), and 52% (P≤0.025***), respectively.</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161235, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g003", "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cell_apoptosis_in_the_presence_of_O_DDHSL_/1161235", "title"=>"Cell apoptosis in the presence of O-DDHSL.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1661558"], "description"=>"<p><b>A</b>- Colony forming assay was performed by plating 2×10<sup>4</sup> cells per plate with or without treatment of Panc-1, Aspc-1 and HPDE cells with O-DDHSL (150 µM) for 48 h. After seven to ten days incubation at 37°C the cells were fixed with 70% methanol and stained with 0.5% crystal violet. Cell colonies were counted using Open CFU software <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0106480#pone.0106480-Geissmann1\" target=\"_blank\">[21]</a>. Colonies, defined as groups of ≥25 cells, were identified and classified as a single colony. <b>B</b> – Relative colony formation depicted as a percentage between O-DDHSL untreated and treated cells, Panc-1(P≤0.029*), Aspc-1(P≤0.03**) and HPDE (P≤0.05***).</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161238, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g004", "stats"=>{"downloads"=>29, "page_views"=>121, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Colony_formation_upon_treatment_with_O_DDHSL_/1161238", "title"=>"Colony formation upon treatment with O-DDHSL.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1661561"], "description"=>"<p><b>A</b>- Panc-1, Aspc-1 and HPDE cells were plated and allowed to grow as a monolayer after which a similar sized scratch was made with a sterile tip in the culture plates. The plates were photographed at baseline (0 h). A set of plates were treated with O-DDHSL (150 µM) and the gap closure was photographed after 48 h. In the presence of O-DDHSL, the migration of cells was considerably less, which resulted in non-closure of the wound gap compared to untreated cells (n = 3). Red arrows - Wound gap (20×). <b>B</b> - The wound area in the image was measured using Image J software and the baseline gap area was assigned an arbitrary number of 1. The untreated carcinoma cell wound closure values at 48 h (P = 0.027*) was significant. The O-DDHSL treated cell wound closure value at 48 h for both Pan-1 and Aspc-1 was not significant (P≥0.05). In HPDE cells, the wound closure between treated and untreated cells was almost similar. <b>C</b> – Detection of a fluorescent analog of O-DDHSL, N-Dd-HSL-3-HF, 10 µM, in different cells after 30 min at 37°C (40×).</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161241, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g005", "stats"=>{"downloads"=>4, "page_views"=>112, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cell_migration_by_wound_healing_assay_/1161241", "title"=>"Cell migration by wound healing assay.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1661562"], "description"=>"<p>In order to analyze the changes in mRNA expression of <i>RhoC</i>, <i>cofilin</i> and <i>IQGAP-1</i> in Panc-1, Aspc-1 and HPDE cells, upon treatment with O-DDHSL (150 µM; 48 h), a qRT-PCR was performed with specific primers. <i>β-actin</i> was used as a housekeeping gene. Basal expression of all the above genes was observed in HPDE cells. <i>Cofilin</i> mRNA level increased in Panc-1 (P = 0.04*) and HPDE cells (P = 0.037*). Similarly, the RhoC mRNA levels increased in Panc-1 (P = 0.032**) and HPDE cells (P = 0.029**), respectively. The <i>IQGAP-1</i> mRNA level decreased in HPDE cells (P = 0.024***). In Aspc-1 cells, no change was observed in <i>IQGAP-1</i> mRNA, but a marginal decrease in <i>cofilin</i> and two fold (P = 0.036**) increase in <i>RhoC</i> mRNA was observed.</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161242, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g006", "stats"=>{"downloads"=>7, "page_views"=>70, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_qRT_PCR_analysis_of_genes_Rho_C_Cofilin_and_IQGAP_1_in_different_cells_/1161242", "title"=>"qRT-PCR analysis of genes <i>Rho C, Cofilin</i> and <i>IQGAP-1</i> in different cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1661563"], "description"=>"<p>Equal amounts of protein (∼40 µg) was separated on a 10% SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. The membrane was probed with primary antibodies for Rho-C (∼42 kDa), cofilin (∼18 kDa), <i>IQGAP-1 (∼197 </i>k<i>DA)</i> and, β-actin (∼47 kDA) followed by incubation with a HRP-conjugated secondary antibody. The bands were detected using a chemiluminescent reagent. The blot image was acquired using a Kodak imaging software.</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161243, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g007", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blot_for_protein_expression_/1161243", "title"=>"Western blot for protein expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1661549"], "description"=>"<p>Chemical structures of homoserinelactones.</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161229, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Chemical_structures_of_homoserinelactones_/1161229", "title"=>"Chemical structures of homoserinelactones.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1661551"], "description"=>"<p><b>A</b>- Panc-1 (6×10<sup>3</sup> cells per well) was treated with different concentrations of O-DDHSL for 24 and 48 h. Significant decrease in cell viability was observed with 200–300 µM O-DDHSL (P≤0.05) after 24 h. At 48 h, cell viability decrease was significant at O-DDHSL concentration at or above 100 µM (P≤0.02; n = 3). However, HPDE cell viability was not affected after 48 h except for a slight decrease at 300 µM O-DDHSL. No effect was observed with O-HHSL. <b>B</b> – Aspc-1 (6×10<sup>3</sup> cells per well) was more sensitive to O-DDHSL exposure than Panc-1. A significant decrease (P≤0.02) was observed in viability ≥25 µM O-DDHSL after 24 or 48 h (n = 3). Cell viability was not affected by O-HHSL. In both cases, DMSO (0.02%) was used as diluent control which did not affect the cell viability <i>per se</i>. <b>C–E</b> Panc-1, Aspc-1 and HPDE cells were plated on a layer of matrigel in chamber slides in serum free DMEM/F12 media and allowed to grow for two weeks. O-DDHSL (200 µM) was added to the cells and incubated for 48 h at 37°C. Subsequently, a fluorescent dye Calcein AM was added and further incubated for 2 h for its uptake by the cells. C–E – Light microsopy pictures of cells growing on matrigel before and after addition of O-DDHSL showing morphological changes of apoptosis (top panel). Bottom panel shows the uptake of Calcein AM dye in the cells before and after treatment with O-DDHSL, showing dye uptake by viable cells and loss of cell viability resulting in the absence of dye uptake (40×).</p>", "links"=>[], "tags"=>["iqgap", "pancreatic carcinoma cells", "Cofilin mRNA expression", "Pancreatic cancer", "hpde", "Pseudomonas aeruginosa quorum", "tumor cell migration", "Human Pancreatic Carcinoma Cells", "cell viability", "pancreatic carcinoma cell lines", "Reduced Cell Motility", "matrigel basement membrane matrix"], "article_id"=>1161231, "categories"=>["Biological Sciences"], "users"=>["Ashwath S. Kumar", "Jeffrey N. Bryan", "Senthil R. Kumar"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106480.g002", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_HSL_on_cell_viability_/1161231", "title"=>"Effect of HSL on cell viability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-04 03:03:43"}

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