Toll-Like Receptor 4 Prompts Human Breast Cancer Cells Invasiveness via Lipopolysaccharide Stimulation and Is Overexpressed in Patients with Lymph Node Metastasis
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{"title"=>"Toll-like receptor 4 prompts human breast cancer cells invasiveness via lipopolysaccharide stimulation and is overexpressed in patients with lymph node metastasis", "type"=>"journal", "authors"=>[{"first_name"=>"Huan", "last_name"=>"Yang", "scopus_author_id"=>"57198825456"}, {"first_name"=>"Bo", "last_name"=>"Wang", "scopus_author_id"=>"57199068698"}, {"first_name"=>"Tao", "last_name"=>"Wang", "scopus_author_id"=>"57199031265"}, {"first_name"=>"Longjiang", "last_name"=>"Xu", "scopus_author_id"=>"56456811500"}, {"first_name"=>"Chunyan", "last_name"=>"He", "scopus_author_id"=>"55970995000"}, {"first_name"=>"Huiyan", "last_name"=>"Wen", "scopus_author_id"=>"36238594300"}, {"first_name"=>"Jie", "last_name"=>"Yan", "scopus_author_id"=>"57199790214"}, {"first_name"=>"Honghong", "last_name"=>"Su", "scopus_author_id"=>"56379474700"}, {"first_name"=>"Xueming", "last_name"=>"Zhu", "scopus_author_id"=>"7406186645"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84907816018", "sgr"=>"84907816018", "pui"=>"600116381", "pmid"=>"25299052", "doi"=>"10.1371/journal.pone.0109980"}, "id"=>"f2c02c70-288a-307b-a3ce-9ae81fd89ca5", "abstract"=>"Toll-like receptor (TLR)4-mediated signaling has been implicated in tumor cell invasion, survival, and metastasis in a variety of cancers. This study investigated the expression and biological role of TLR4 in human breast cancer metastasis. MCF-7 and MDA-MB-231 are human breast cancer cell lines with low and high metastatic potential, respectively. Using lipopolysaccharide (LPS) to stimulate MCF-7 and MDA-MB-231 cells, expression of TLR4 mRNA and protein increased compared with that in control cells. TLR4 activation notably up-regulated expression of matrix metalloproteinase (MMP)-2, MMP-9 and vascular endothelial growth factor(VEGF) mRNA and their secretion in the supernatants of both cell lines. LPS enhanced invasion of MDA-MB-231 cells by transwell assay and MCF-7 cells by wound healing assay. LPS triggered increased expression of TLR4 downstream signaling pathway protein myeloid differentiation factor 88(MyD88) and resulted in interleukin (IL)-6 and IL-10 higher production by human breast cancer cells. Stimulation of TLR4 with LPS promoted tumorigenesis and formed metastatic lesions in liver of nude mice. Moreover, expression of TLR4 and MyD88 as well as invasiveness and migration of the cells could be blocked by TLR4 antagonist. Combined with clinicopathological parameters, TLR4 was overexpressed in human breast cancer tissue and correlated with lymph node metastasis. These findings indicated that TLR4 may participate in the progression and metastasis of human breast cancer and provide a new therapeutic target.", "link"=>"http://www.mendeley.com/research/tolllike-receptor-4-prompts-human-breast-cancer-cells-invasiveness-via-lipopolysaccharide-stimulatio", "reader_count"=>47, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Student > Doctoral Student"=>3, "Researcher"=>6, "Student > Ph. D. Student"=>15, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>3, "Student > Bachelor"=>6, "Lecturer"=>2, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Student > Doctoral Student"=>3, "Researcher"=>6, "Student > Ph. D. Student"=>15, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>3, "Student > Bachelor"=>6, "Lecturer"=>2, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>11, "Medicine and Dentistry"=>10, "Agricultural and Biological Sciences"=>18, "Pharmacology, Toxicology and Pharmaceutical Science"=>3, "Chemistry"=>1, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>11}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>3}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1711596"], "description"=>"<p>(A) mRNA of MMP-2, MMP-9 and VEGF were analyzed by RT-PCR in control and LPS (2 µg/ml, 48 h) groups. (B) mRNA of MMP-2, MMP-9 and VEGF were analyzed by real-time PCR in control and LPS groups(2 µg/ml, 48 h). Each bar represented triplicate analyses of mean±SD, **P<0.05. (C) MMP-2, MMP-9 and VEGF in the culture supernatants of control and LPS groups (0, 2, 10 and 20 µg/ml) for 48 h was measured by ELISA. Each bar represented triplicate analyses of mean±SD, **P<0.05.</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199232, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g002", "stats"=>{"downloads"=>17, "page_views"=>267, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LPS_upregulated_MMP_2_MMP_9_and_VEGF_production_by_MCF_7_and_MDA_MB_231_cells_/1199232", "title"=>"LPS upregulated MMP-2, MMP-9 and VEGF production by MCF-7 and MDA-MB-231 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711630"], "description"=>"<p>(A) Immunohistochemistry for TLR4 in tissue sections. Magnification, ×200. (a) TLR4 expression in normal breast tissue. (b) TLR4 expression in lymph node metastasis (N0) breast cancer tissue. (c) TLR4 expression in lymph node metastasis (N1) breast cancer tissue. (d) TLR4 expression in lymph node metastasis (N2) breast cancer tissue. (e) TLR4 expression in lymph node metastasis (N3) breast cancer tissue. (B) mRNA of TLR4 in normal breast tissue and tumor tissue by RT-PCR. (C) mRNA of TLR4 in normal breast tissue and tumor tissue by real-time PCR. Each bar represents analyses of mean±SD, **P<0.05.</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199264, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g008", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TLR4_expression_in_clinical_tissue_/1199264", "title"=>"TLR4 expression in clinical tissue.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711598"], "description"=>"<p>(A) Matrigel invasion assay was performed to observe the changes in MDA-MB-231 cell invasion. The LPS group was stimulated with 10 µg/ml LPS for 48 h. Magnification, ×200. (B) Total number of invading cells through the matrigel was counted under the microscope in the whole fields. Each bar represents the mean±SD of three wells counted. **P<0.05. Results are representative of three separate experiments. (C) Wound-healing assay was performed to observe the changes in MCF-7 cell invasion after stimulation. The LPS group was also stimulated with 10 µg/ml LPS for 48 h. Magnification, ×100. All results were representative of three separate experiments.</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199234, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g003", "stats"=>{"downloads"=>1, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Stimulation_of_TLR4_promoted_migration_and_invasion_of_breast_cancer_cell_lines_/1199234", "title"=>"Stimulation of TLR4 promoted migration and invasion of breast cancer cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711601"], "description"=>"<p>(A) Expression of MyD88 was analyzed by western blotting. LPS stimulation (2 µg/ml) for 48 h induced higher expression of MyD88 in both cell lines. (B) Representative images of IL-6 and IL-10 in supernatants from control and LPS groups (20 µg/ml, 48 h) of both cell lines measured by flow cytometry. (C) and (D) Quantitative analysis of cytokines in supernatants of MCF-7 and MDA-MB-231 cells treated with LPS (0, 2, 10 and 20 µg/ml) for 48 h. Each bar represents triplicate analyses of mean±SD, **P<0.05. All results were representative of three separate experiments.</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199237, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g004", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LPS_promoted_MyD88_IL_6_and_IL_10_production_by_human_breast_cancer_cells_/1199237", "title"=>"LPS promoted MyD88, IL-6 and IL-10 production by human breast cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711631", "https://ndownloader.figshare.com/files/1711632"], "description"=>"<div><p>Toll-like receptor (TLR)4-mediated signaling has been implicated in tumor cell invasion, survival, and metastasis in a variety of cancers. This study investigated the expression and biological role of TLR4 in human breast cancer metastasis. MCF-7 and MDA-MB-231 are human breast cancer cell lines with low and high metastatic potential, respectively. Using lipopolysaccharide (LPS) to stimulate MCF-7 and MDA-MB-231 cells, expression of TLR4 mRNA and protein increased compared with that in control cells. TLR4 activation notably up-regulated expression of matrix metalloproteinase (MMP)-2, MMP-9 and vascular endothelial growth factor(VEGF) mRNA and their secretion in the supernatants of both cell lines. LPS enhanced invasion of MDA-MB-231 cells by transwell assay and MCF-7 cells by wound healing assay. LPS triggered increased expression of TLR4 downstream signaling pathway protein myeloid differentiation factor 88(MyD88) and resulted in interleukin (IL)-6 and IL-10 higher production by human breast cancer cells. Stimulation of TLR4 with LPS promoted tumorigenesis and formed metastatic lesions in liver of nude mice. Moreover, expression of TLR4 and MyD88 as well as invasiveness and migration of the cells could be blocked by TLR4 antagonist. Combined with clinicopathological parameters, TLR4 was overexpressed in human breast cancer tissue and correlated with lymph node metastasis. These findings indicated that TLR4 may participate in the progression and metastasis of human breast cancer and provide a new therapeutic target.</p></div>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199265, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0109980.s001", "https://dx.doi.org/10.1371/journal.pone.0109980.s002"], "stats"=>{"downloads"=>11, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Toll_Like_Receptor_4_Prompts_Human_Breast_Cancer_Cells_Invasiveness_via_Lipopolysaccharide_Stimulation_and_Is_Overexpressed_in_Patients_with_Lymph_Node_Metastasis_/1199265", "title"=>"Toll-Like Receptor 4 Prompts Human Breast Cancer Cells Invasiveness via Lipopolysaccharide Stimulation and Is Overexpressed in Patients with Lymph Node Metastasis", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711606"], "description"=>"<p>(A) Comparison of breast tumor volume from LPS treatment group. The mean±SD tumor volume of five animals per group. **P<0.05. (B) Comparison of breast tumor weight from LPS treatment group. Each bar represents the mean±SD tumor weight of five animals per group. **P<0.05. (C) TLR4 expression in transplanted tumor sections. Magnification,×200. (a) and (b) Immunohistochemistry for TLR4 in transplanted tumor sections. (a) is control group and (b) is LPS group. (c) H&E staining of the liver from control group. (d) H&E staining of the liver from LPS group. Magnification,×100.</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199242, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g005", "stats"=>{"downloads"=>3, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TLR4_stimulation_with_LPS_promoted_tumorigenesis_in_the_tumor_model_of_MDA_MB_231_/1199242", "title"=>"TLR4 stimulation with LPS promoted tumorigenesis in the tumor model of MDA-MB-231.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711609"], "description"=>"<p>(A)Expression of TLR4 was analyzed by real-time PCR. **P<0.05. Each bar represented triplicate analyses of mean±SD. (B)Expression of TLR4, MyD88 was analyzed by western blotting. All results were representative of three separate experiments. LPS (2 µg/ml, 48 h), ER(100 nmol/L eritoran, 30 min pretreatment, 48 h), ER+LPS(100 nmol/L eritoran, 30 min prior to 2 µg/ml LPS, 48 h).</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199245, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g006", "stats"=>{"downloads"=>3, "page_views"=>150, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_TLR4_antagonist_eritoran_on_LPS_induced_expression_of_TLR4_and_MyD88_in_human_breast_cancer_cells_/1199245", "title"=>"Effects of TLR4 antagonist eritoran on LPS-induced expression of TLR4 and MyD88 in human breast cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711623"], "description"=>"<p>(A) mRNA of MMP-2, MMP-9 and VEGF were analyzed by real-time PCR. Each bar represented triplicate analyses of mean±SD, **P<0.05. LPS (2 µg/ml, 48 h), ER(100 nmol/L eritoran,30 min pretreatment, 48 h), ER+LPS(100 nmol/L eritoran,30 min prior to 2 µg/ml LPS, 48 h).(B) Invasiveness of MDA-MB-231 cells was performed by transwell assay. (a)Control,(b)LPS,(c)ER,(d)ER+LPS. Magnification, ×200. (e) Total number of invading cells through the matrigel was counted under the microscope in the whole fields. Each bar represents the mean±SD of three wells counted. **P<0.05. Results are representative of three separate experiments. (C) Wound-healing assay was performed to observe the changes in MCF-7 cells invasion. Magnification, ×100. All results were representative of three separate experiments. (B),(C) LPS (10 µg/ml, 48 h), ER(100 nmol/L eritoran,30 min pretreatment, 48 h), ER+LPS(100 nmol/L eritoran,30 min prior to 10 µg/ml LPS, 48 h).</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199257, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g007", "stats"=>{"downloads"=>5, "page_views"=>36, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TLR4_antagonist_blocked_invasiveness_and_migration_in_human_breast_cancer_cells_/1199257", "title"=>"TLR4 antagonist blocked invasiveness and migration in human breast cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711593"], "description"=>"<p>(A) Expression of TLR4 in control and stimulated (2 µg/ml LPS) MCF-7 and MDA-MB-231 cells for 48 h by RT-PCR. (B) Expression of TLR4 in control and LPS groups(2 µg/ml, 48 h) was analyzed by real-time PCR, **P<0.05. Each bar represented triplicate analyses of mean±SD. (C) Expression of TLR4 in control and LPS groups (2 µg/ml, 48 h)was analyzed by flow cytometry,**P<0.05. Each bar represented triplicate analyses of mean±SD. (D) Expression of TLR4 in control and LPS groups(2 µg/ml, 48 h) was analyzed by western blotting. All results were representative of three separate experiments.</p>", "links"=>[], "tags"=>["TLR 4 antagonist", "lps", "tumor cell invasion", "TLR 4", "wound healing assay", "TLR 4 activation", "expression", "breast cancer cells", "breast cancer cell lines", "mmp", "lymph node metastasis", "breast cancer tissue", "breast cancer metastasis", "il", "mcf", "TLR 4 mRNA"], "article_id"=>1199230, "categories"=>["Biological Sciences"], "users"=>["Huan Yang", "Bo Wang", "Tao Wang", "Longjiang Xu", "Chunyan He", "Huiyan Wen", "Jie Yan", "Honghong Su", "Xueming Zhu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0109980.g001", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_TLR4_in_human_breast_cancer_cells_/1199230", "title"=>"Expression of TLR4 in human breast cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:30:58"}

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