title: Macrophages and endothelial cells orchestrate tumor-associated angiogenesis in oral cancer via hedgehog pathway activation.
abstract: The present study aimed to evaluate the role of Hedgehog (Hh) molecule expression in association with the clinical aspects of oral squamous cell carcinoma (OSCC), as well as angiogenesis and CD163+ macrophages. Twenty-eight cases of OSCC, nine cases of tumor-free resection margins (TM), and four cases of non-neoplastic oral mucosa (NNM) were submitted to immunohistochemistry to detect proteins Sonic Hedgehog (SHH), Indian Hedgehog (IHH), GLI1, CD163, and CD105. Protein colocalization with respect to SHH/CD163, IHH/CD163, GLI1/CD163, and GLI1/CD105 was assessed by immunohistochemical double staining. In tumor parenchyma, SHH and IHH were present in the cytoplasm of neoplastic cells, while GLI1 was observed in cytoplasm and nucleus. Endothelial cells were found to express SHH, IHH, and GLI1 within CD105+ vessels, and a positive correlation between infiltrating macrophage density (IMD) and microvascular density (MVD) was observed in cases of OSCC and TM. When compared to TM and NNM, the OSCC cases demonstrated higher immunoreactivity for SHH (p = 0.01), IHH (p = 0.39), GLI1 (p = 0.03), IMD (p = 0.0002), and MVD (p = 0.0002). Our results suggest the participation of the Hh pathway in OSCC by way of autocrine and paracrine signaling, in addition to the participation of both SHH and IHH ligands. Endothelial cells were also found to exhibit positivity with respect to Hh pathway components and we surmise that these molecules may play a role in tumor angiogenesis. CD163+ macrophages were also observed to express IHH, a ligand of this pathway, in addition to being associated with tumor neovascularization.
authors.affiliation: Laboratory of Pathology and Molecular Biology, Oswaldo Cruz Foundation, Gonçalo Moniz Research Center, R. Waldemar Falcão, 121, Candeal, Salvador, Bahia, CEP: 40296-710, Brazil.
authors.forename: Ludmila de Faro
authors.initials: Lde F
authors.lastname: Valverde
authors.affiliation: Laboratory of Experimental Pathology, Oswaldo Cruz Foundation, Salvador, Bahia, Brazil.
authors.forename: Thiago de Almeida
authors.initials: Tde A
authors.lastname: Pereira
authors.affiliation: Laboratory of Pathology and Molecular Biology, Oswaldo Cruz Foundation, Gonçalo Moniz Research Center, R. Waldemar Falcão, 121, Candeal, Salvador, Bahia, CEP: 40296-710, Brazil.
authors.forename: Rosane Borges
authors.initials: RB
authors.lastname: Dias
authors.affiliation: Laboratory of Pathology and Molecular Biology, Oswaldo Cruz Foundation, Gonçalo Moniz Research Center, R. Waldemar Falcão, 121, Candeal, Salvador, Bahia, CEP: 40296-710, Brazil.
authors.forename: Vanessa Sousa Nazaré
authors.initials: VS
authors.lastname: Guimarães
authors.affiliation: Laboratory of Pathology and Molecular Biology, Oswaldo Cruz Foundation, Gonçalo Moniz Research Center, R. Waldemar Falcão, 121, Candeal, Salvador, Bahia, CEP: 40296-710, Brazil.
authors.forename: Eduardo Antônio Gonçalves
authors.initials: EA
authors.lastname: Ramos
authors.affiliation: Laboratory of Oral Surgical Pathology, School of Dentistry of the Federal University of Bahia, Salvador, Bahia, Brazil.
authors.forename: Jean Nunes
authors.initials: JN
authors.lastname: Santos
authors.affiliation: Laboratory of Pathology and Molecular Biology, Oswaldo Cruz Foundation, Gonçalo Moniz Research Center, R. Waldemar Falcão, 121, Candeal, Salvador, Bahia, CEP: 40296-710, Brazil. gurgel.clarissa@gmail.com.|Laboratory of Oral Surgical Pathology, School of Dentistry of the Federal University of Bahia, Salvador, Bahia, Brazil. gurgel.clarissa@gmail.com.
authors.forename: Clarissa Araújo
authors.initials: CA
authors.lastname: Gurgel Rocha
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D000071063:Endoglin / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D053823:Hedgehog Proteins / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000473:pathology*; D008297:Male; D009061:Mouth Mucosa / Q000378:metabolism / Q000473:pathology; D009062:Mouth Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D011956:Receptors, Cell Surface / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D000071676:Zinc Finger Protein GLI1 / Q000378:metabolism; D000098902:CD163 Antigen
chemical_list: D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D000098902:CD163 Antigen; D000071063:Endoglin; D053823:Hedgehog Proteins; C509246:IHH protein, human; D011956:Receptors, Cell Surface; C509239:SHH protein, human; D000071676:Zinc Finger Protein GLI1
keywords: Hedgehog proteins; Immunohistochemistry; Oral cancer; Squamous cell carcinoma