Cancer-Associated Fibroblasts in a Human HEp-2 Established Laryngeal Xenografted Tumor Are Not Derived from Cancer Cells through Epithelial-Mesenchymal Transition, Phenotypically Activated but Karyotypically Normal
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{"title"=>"Cancer-associated fibroblasts in a human HEp-2 established laryngeal xenografted tumor are not derived from cancer cells through epithelial-mesenchymal transition, phenotypically activated but karyotypically normal", "type"=>"journal", "authors"=>[{"first_name"=>"Mei", "last_name"=>"Wang", "scopus_author_id"=>"56177053500"}, {"first_name"=>"Chun Ping", "last_name"=>"Wu", "scopus_author_id"=>"55763317500"}, {"first_name"=>"Jun Yan", "last_name"=>"Pan", "scopus_author_id"=>"56510749200"}, {"first_name"=>"Wen Wei", "last_name"=>"Zheng", "scopus_author_id"=>"56510748800"}, {"first_name"=>"Xiao Juan", "last_name"=>"Cao", "scopus_author_id"=>"56510722100"}, {"first_name"=>"Guo Kang", "last_name"=>"Fan", "scopus_author_id"=>"7202776046"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"25658113", "sgr"=>"84922553309", "doi"=>"10.1371/journal.pone.0117405", "scopus"=>"2-s2.0-84922553309", "pui"=>"602121528", "isbn"=>"1932-6203", "issn"=>"19326203"}, "id"=>"5d8e2fee-522b-3dd4-8ad9-4e29115c7741", "abstract"=>"Cancer-associated fibroblasts (CAFs) play a crucial role in cancer progression and even initiation. However, the origins of CAFs in various cancer types remain controversial, and one of the important hypothesized origins is through epithelial-mesenchymal transition (EMT) from cancer cells. In this study, we investigated whether the HEp-2 laryngeal cancer cells are able to generate CAFs via EMT during tumor formation, which is now still unknown. The laryngeal xenografted tumor model was established by inoculating the HEp-2 laryngeal cancer cell line in nude mice. Primary cultured CAFs from the tumor nodules and matched normal fibroblasts (NFs) from the adjacent connective tissues were subcultured, purified, and verified by immunofluorescence. Migration, invasion, and proliferation potentials were compared between the CAFs and NFs. A co-culture of CAFs with HEp-2 cells and a co-injection of CAFs with HEp-2 cells in nude mice were performed to examine the cancer-promoting potential of CAFs to further verify their identity. Karyotypic analyses of the CAFs, NFs, and HEp-2 cells were conducted. A co-culture of NFs with HEp-2 cells was also performed to examine the expression of activated markers of CAFs. A pathological examination confirmed that the laryngeal xenografted tumor model was successfully established, containing abundant CAFs. Immunocytochemical staining verified the purities and identities of the CAFs and NFs. Although the CAFs manifested higher migration, invasion, proliferation, and cancer-promoting capacities compared with the NFs, an analysis of chromosomes revealed that both the CAFs and NFs showed typical normal mouse karyotypes. In addition, the NFs co-cultured with HEp-2 cells did not show induced expressions of activated markers of CAFs. Our findings reveal that the CAFs in the HEp-2 established laryngeal xenografted tumor are not of laryngeal cancer origin but of mouse origin, indicating that the HEp-2 laryngeal cancer cells cannot generate their own CAFs via EMT in this model.", "link"=>"http://www.mendeley.com/research/cancerassociated-fibroblasts-human-hep2-established-laryngeal-xenografted-tumor-not-derived-cancer-c", "reader_count"=>9, "reader_count_by_academic_status"=>{"Researcher"=>3, "Student > Ph. D. Student"=>2, "Student > Bachelor"=>3, "Student > Doctoral Student"=>1}, "reader_count_by_user_role"=>{"Researcher"=>3, "Student > Ph. D. Student"=>2, "Student > Bachelor"=>3, "Student > Doctoral Student"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Materials Science"=>1, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>2, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1885325"], "description"=>"<p>(A,B) Wound healing assay and statistical analysis. The HEp-2 cells co-cultured with CAFs closed the gaps significantly faster compared with the control cells at time points of 24, 48, and 72 h. (C,D) Invasion assay and statistical analysis. The HEp-2 cells co-cultured with NFs had similar invasion capacity compared with those HEp-2 cells cultured alone. By contrast, the HEp-2 cells co-cultured with CAFs showed significantly higher invasion capacity than the control cells. (E) Cell growth rate measured by cell counting revealed that the co-cultured CAFs promoted the proliferation capacity of HEp-2 cells more than the NFs did (<sup>n.s.</sup>p>0.05, *p<0.05, **p<0.01).</p>", "links"=>[], "tags"=>["cancer progression", "laryngeal cancer origin", "tumor formation", "tumor nodules", "proliferation potentials", "cancer types", "cancer cells", "emt", "laryngeal xenografted tumor model", "Phenotypically Activated", "laryngeal xenografted tumor", "mouse karyotypes", "mouse origin", "nf", "Immunocytochemical staining", "Karyotypic analyses", "hep", "caf"], "article_id"=>1303968, "categories"=>["Biological Sciences"], "users"=>["Mei Wang", "Chun-Ping Wu", "Jun-Yan Pan", "Wen-Wei Zheng", "Xiao-Juan Cao", "Guo-Kang Fan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117405.g004", "stats"=>{"downloads"=>5, "page_views"=>73, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Co_cultured_CAFs_promoted_the_migration_invasion_and_proliferation_potentials_of_HEp_2_cells_/1303968", "title"=>"Co-cultured CAFs promoted the migration, invasion, and proliferation potentials of HEp-2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-06 06:16:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1885310"], "description"=>"<p>(A) Four weeks after injection, tumor nodules formed in the right armpits of nude mice (black arrows). (B–D) HE staining confirmed that the nodules were typical laryngeal squamous cell carcinoma containing abundant CAFs (black arrows). (E,F) Two days after seeding, the CAFs (white arrow) and HEp-2 cells (black arrow) grew out of the tissue fragments. (G,H) At passage 4, purified CAFs were obtained. (I,J) The NFs grew out of the fragment. (K,L) At passage 4, purified NFs were obtained.</p>", "links"=>[], "tags"=>["cancer progression", "laryngeal cancer origin", "tumor formation", "tumor nodules", "proliferation potentials", "cancer types", "cancer cells", "emt", "laryngeal xenografted tumor model", "Phenotypically Activated", "laryngeal xenografted tumor", "mouse karyotypes", "mouse origin", "nf", "Immunocytochemical staining", "Karyotypic analyses", "hep", "caf"], "article_id"=>1303953, "categories"=>["Biological Sciences"], "users"=>["Mei Wang", "Chun-Ping Wu", "Jun-Yan Pan", "Wen-Wei Zheng", "Xiao-Juan Cao", "Guo-Kang Fan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117405.g001", "stats"=>{"downloads"=>2, "page_views"=>43, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Tumor_formation_in_nude_mice_HE_staining_and_primary_culture_of_CAFs_and_NFs_/1303953", "title"=>"Tumor formation in nude mice, HE staining, and primary culture of CAFs and NFs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-06 06:16:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1885327"], "description"=>"<p>Both the CAFs (A,B) and NFs (C,D) showed typical characteristics of normal male mouse karyotype: acrocentric chromosome, modal number of 40 (38+XY), and without numerical or structural aberrations. (E,F) In contrast, the HEp-2 cell line showed a near-triploid karyotype, with both numerical and structural aberrations. Three marker chromosomes were consistently observed.</p>", "links"=>[], "tags"=>["cancer progression", "laryngeal cancer origin", "tumor formation", "tumor nodules", "proliferation potentials", "cancer types", "cancer cells", "emt", "laryngeal xenografted tumor model", "Phenotypically Activated", "laryngeal xenografted tumor", "mouse karyotypes", "mouse origin", "nf", "Immunocytochemical staining", "Karyotypic analyses", "hep", "caf"], "article_id"=>1303970, "categories"=>["Biological Sciences"], "users"=>["Mei Wang", "Chun-Ping Wu", "Jun-Yan Pan", "Wen-Wei Zheng", "Xiao-Juan Cao", "Guo-Kang Fan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117405.g005", "stats"=>{"downloads"=>1, "page_views"=>37, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Representative_karyotypes_of_CAFs_NFs_and_HEp_2_cells_/1303970", "title"=>"Representative karyotypes of CAFs, NFs, and HEp-2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-06 06:16:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1885328"], "description"=>"<p>(A,B) Protein levels of α-SMA and Fap in NFs after being co-cultured with HEp-2 cells for 7 d. The NFs and CAFs served as controls. The HEp-2 co-cultured NFs did not show elevated expressions of α-SMA and Fap. (C) Tumor nodules induced by CAFs and NFs (both mixed with HEp-2 cells) in nude mice. (D) Pathological examination revealed that all tumor nodules were typical laryngeal squamous cell carcinoma, and the tumor cells were surrounded by numerous CAFs. (E) Statistical analysis of tumor weights (**p<0.01).</p>", "links"=>[], "tags"=>["cancer progression", "laryngeal cancer origin", "tumor formation", "tumor nodules", "proliferation potentials", "cancer types", "cancer cells", "emt", "laryngeal xenografted tumor model", "Phenotypically Activated", "laryngeal xenografted tumor", "mouse karyotypes", "mouse origin", "nf", "Immunocytochemical staining", "Karyotypic analyses", "hep", "caf"], "article_id"=>1303973, "categories"=>["Biological Sciences"], "users"=>["Mei Wang", "Chun-Ping Wu", "Jun-Yan Pan", "Wen-Wei Zheng", "Xiao-Juan Cao", "Guo-Kang Fan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117405.g006", "stats"=>{"downloads"=>1, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blot_and_tumor_promoting_capacity_of_CAFs_and_NFs_/1303973", "title"=>"Western blot and tumor-promoting capacity of CAFs and NFs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-06 06:16:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1885321"], "description"=>"<p>(A,B) Gap closure assay and statistical analysis. The CAFs showed higher migration capacity compared with NFs and completely closed the gaps within 48 h. (C,D) Invasion assay and statistical analysis. The CAFs were found to be able to invade across the Matrigel; however, no NFs were found to invade across the Matrigel, indicating activated invasion potential of CAFs. (E) Cell growth rate measured by CCK-8 assay revealed that the CAFs had higher proliferation capacity than the NFs did. (F) Growth rate difference of CAFs in different passages. The CAFs began to show increasingly decreased proliferation capacity from passage 15 (<sup>n.s.</sup>p>0.05, *p<0.05, **p<0.01).</p>", "links"=>[], "tags"=>["cancer progression", "laryngeal cancer origin", "tumor formation", "tumor nodules", "proliferation potentials", "cancer types", "cancer cells", "emt", "laryngeal xenografted tumor model", "Phenotypically Activated", "laryngeal xenografted tumor", "mouse karyotypes", "mouse origin", "nf", "Immunocytochemical staining", "Karyotypic analyses", "hep", "caf"], "article_id"=>1303964, "categories"=>["Biological Sciences"], "users"=>["Mei Wang", "Chun-Ping Wu", "Jun-Yan Pan", "Wen-Wei Zheng", "Xiao-Juan Cao", "Guo-Kang Fan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117405.g003", "stats"=>{"downloads"=>2, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Isolated_CAFs_showed_higher_migration_invasion_and_proliferation_potentials_than_the_NFs_/1303964", "title"=>"Isolated CAFs showed higher migration, invasion, and proliferation potentials than the NFs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-06 06:16:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1885314"], "description"=>"<p>(A,B) Cell morphology of CAFs (passage 25) and NFs (passage 13). The CAFs did not show significant morphological changes compared with the NFs before passage 25; however, the NFs showed changes in morphology, including an enlarged and more flattened outline, compared with the CAFs after passage 10. (C) Cell morphology of primary cultured mouse skin epithelia at passage 1. (D) The mouse skin epithelia showed positive staining of pan-CK (positive control). (E–L) The CAFs and NFs showed negative staining of pan-CK and positive staining of vimentin, indicating their fibroblast identities. Compared with the NFs, the CAFs showed positive staining of α-SMA and Fap, two markers of activated fibroblasts.</p>", "links"=>[], "tags"=>["cancer progression", "laryngeal cancer origin", "tumor formation", "tumor nodules", "proliferation potentials", "cancer types", "cancer cells", "emt", "laryngeal xenografted tumor model", "Phenotypically Activated", "laryngeal xenografted tumor", "mouse karyotypes", "mouse origin", "nf", "Immunocytochemical staining", "Karyotypic analyses", "hep", "caf"], "article_id"=>1303957, "categories"=>["Biological Sciences"], "users"=>["Mei Wang", "Chun-Ping Wu", "Jun-Yan Pan", "Wen-Wei Zheng", "Xiao-Juan Cao", "Guo-Kang Fan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117405.g002", "stats"=>{"downloads"=>6, "page_views"=>273, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cell_morphology_of_CAFs_NFs_and_mouse_skin_epithelia_and_immunocytochemical_staining_/1303957", "title"=>"Cell morphology of CAFs, NFs, and mouse skin epithelia and immunocytochemical staining.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-06 06:16:00"}

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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