Vocal fold fibroblasts promote angiogenesis in vocal fold leukoplakia by secreting pro-angiogenic factors. 2022

Yinying Chu, and Yi Fang, and Haitao Wu, and Jian Chen, and Lei Cheng
ENT Institute and Department of Otolaryngology-Head and Neck Surgery, Eye & ENT Hospital, Fudan University, 83 FenYang Road, Shanghai 200031, China.

OBJECTIVE Cancer-associated fibroblasts (CAFs) have been reported to play an essential role in tumor angiogenesis and progression. In this study, we aimed to investigate the impact of vocal fold leukoplakia-associated fibroblasts (VFLFs) on the angiogenesis process in vocal fold leukoplakia (VFL) and their potential secretions of proangiogenic factors. METHODS A total of 160 lesions (86 laryngeal carcinoma, 67 vocal fold leukoplakia, 7 vocal fold polyp) were detected under narrow band imaging (NBI) mode to evaluate the relationship between pathology and intraepithelial papillary capillary loop (IPCL) grades. We characterized immortalized vocal fold CAFs, VFLFs, normal fibroblasts (NFs) cell lines using immunofluorescence cytochemistry and real-time quantitative polymerase chain reaction (RT-qPCR). The effects of fibroblast conditioned media (CM) on the proliferative, migrating and tube formation capacity of human umbilical vein endothelial cells (HUVEC) were investigated using the cell counting kit-8 (CCK-8) assay, wound healing assay, transwell migration experiment and Matrigel tube formation experiment. The expression levels of proangiogenic factors in CAFs, VFLFs, and NFs were evaluated by antibody microarray and RT-qPCR. RESULTS NBI images depicted that angiogenesis was abnormally activated during laryngeal tumorigenesis. Both CAF and VFLF expressed Vimentin, alpha-smooth muscle actin (α-SMA) and fibroblast activation protein (FAP). NF expressed Vimentin and α-SMA, but not FAP. The PCR results showed that mRNA expression levels of Vimentin, α-SMA and FAP in CAFs and VFLFs were significantly increased than those in NFs. CAF-CM and VFLF-CM promoted the proliferative, migrating, and tube formation ability of HUVECs. Secretome profiling of fibroblasts by antibody microarray demonstrated that VFLFs secreted significantly more vascular endothelial growth factor (VEGF), angiogenin, bFGF and HGF than NFs. CONCLUSIONS Overall, we demonstrated that VEGF, angiogenin, bFGF and HGF derived from VFLFs may play crucial roles in the angiogenesis process of laryngeal premalignant and malignant lesions. This may contribute to the exploitation of novel therapeutic strategies for VFL.

UI MeSH Term Description Entries
D007822 Laryngeal Neoplasms Cancers or tumors of the LARYNX or any of its parts: the GLOTTIS; EPIGLOTTIS; LARYNGEAL CARTILAGES; LARYNGEAL MUSCLES; and VOCAL CORDS. Cancer of Larynx,Laryngeal Cancer,Larynx Neoplasms,Cancer of the Larynx,Larynx Cancer,Neoplasms, Laryngeal,Cancer, Laryngeal,Cancer, Larynx,Cancers, Laryngeal,Cancers, Larynx,Laryngeal Cancers,Laryngeal Neoplasm,Larynx Cancers,Larynx Neoplasm,Neoplasm, Laryngeal,Neoplasm, Larynx,Neoplasms, Larynx
D007971 Leukoplakia A white patch lesion found on a MUCOUS MEMBRANE that cannot be scraped off. Leukoplakia is generally considered a precancerous condition, however its appearance may also result from a variety of HEREDITARY DISEASES. Leukokeratosis,Leukoplakic Lesions,Lesion, Leukoplakic,Lesions, Leukoplakic,Leukokeratoses,Leukoplakias,Leukoplakic Lesion
D009389 Neovascularization, Pathologic A pathologic process consisting of the proliferation of blood vessels in abnormal tissues or in abnormal positions. Angiogenesis, Pathologic,Angiogenesis, Pathological,Neovascularization, Pathological,Pathologic Angiogenesis,Pathologic Neovascularization,Pathological Angiogenesis,Pathological Neovascularization
D005347 Fibroblasts Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules. Fibroblast
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D014746 Vimentin An intermediate filament protein found in most differentiating cells, in cells grown in tissue culture, and in certain fully differentiated cells. Its insolubility suggests that it serves a structural function in the cytoplasm. MW 52,000.
D014827 Vocal Cords A pair of cone-shaped elastic mucous membrane projecting from the laryngeal wall and forming a narrow slit between them. Each contains a thickened free edge (vocal ligament) extending from the THYROID CARTILAGE to the ARYTENOID CARTILAGE, and a VOCAL MUSCLE that shortens or relaxes the vocal cord to control sound production. Vocal Fold,Vocal Folds,Vocal Ligament,Cord, Vocal,Cords, Vocal,Fold, Vocal,Folds, Vocal,Ligament, Vocal,Ligaments, Vocal,Vocal Cord,Vocal Ligaments
D042461 Vascular Endothelial Growth Factor A The original member of the family of endothelial cell growth factors referred to as VASCULAR ENDOTHELIAL GROWTH FACTORS. Vascular endothelial growth factor-A was originally isolated from tumor cells and referred to as "tumor angiogenesis factor" and "vascular permeability factor". Although expressed at high levels in certain tumor-derived cells it is produced by a wide variety of cell types. In addition to stimulating vascular growth and vascular permeability it may play a role in stimulating VASODILATION via NITRIC OXIDE-dependent pathways. Alternative splicing of the mRNA for vascular endothelial growth factor A results in several isoforms of the protein being produced. Vascular Endothelial Growth Factor,Vascular Endothelial Growth Factor-A,GD-VEGF,Glioma-Derived Vascular Endothelial Cell Growth Factor,VEGF,VEGF-A,Vascular Permeability Factor,Vasculotropin,Glioma Derived Vascular Endothelial Cell Growth Factor,Permeability Factor, Vascular
D042783 Endothelial Cells Highly specialized EPITHELIAL CELLS that line the HEART; BLOOD VESSELS; and lymph vessels, forming the ENDOTHELIUM. They are polygonal in shape and joined together by TIGHT JUNCTIONS. The tight junctions allow for variable permeability to specific macromolecules that are transported across the endothelial layer. Capillary Endothelial Cells,Lymphatic Endothelial Cells,Vascular Endothelial Cells,Capillary Endothelial Cell,Cell, Capillary Endothelial,Cell, Endothelial,Cell, Lymphatic Endothelial,Cell, Vascular Endothelial,Cells, Capillary Endothelial,Cells, Endothelial,Cells, Lymphatic Endothelial,Cells, Vascular Endothelial,Endothelial Cell,Endothelial Cell, Capillary,Endothelial Cell, Lymphatic,Endothelial Cell, Vascular,Endothelial Cells, Capillary,Endothelial Cells, Lymphatic,Endothelial Cells, Vascular,Lymphatic Endothelial Cell,Vascular Endothelial Cell
D043925 Angiogenesis Inducing Agents Agents that induce or stimulate PHYSIOLOGIC ANGIOGENESIS or PATHOLOGIC ANGIOGENESIS. Angiogenesis Inducers,Angiogenesis Effect,Angiogenesis Effects,Angiogenesis Factor,Angiogenesis Stimulating Agents,Angiogenesis Stimulators,Angiogenic Factor,Tumor Angiogenic Factor,Agents, Angiogenesis Inducing,Agents, Angiogenesis Stimulating,Angiogenic Factor, Tumor,Effect, Angiogenesis,Effects, Angiogenesis,Factor, Angiogenesis,Factor, Angiogenic,Factor, Tumor Angiogenic,Inducers, Angiogenesis,Inducing Agents, Angiogenesis,Stimulating Agents, Angiogenesis,Stimulators, Angiogenesis

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