lipofectamine and Laryngeal-Neoplasms

The expression pattern, functions, and detailed regulatory mechanisms of miR-379 in laryngeal carcinoma remain unknown. In the present study, we aimed to uncover novel potential miR-379 targets and shed light on its roles in laryngeal carcinoma. The expression level of miR-379 was measured based on the data obtained from the TCGA database and the cell lines. After miR-379 mimic transfection, cell proliferation, invasion and migration assay, and wound-healing assay in HEp-2 cell line were implemented to evaluate the effects of miR-379 on laryngeal carcinoma in vitro. The target genes for miR-379 in silico were predicted and validated using luciferase reporter assay. The miR-379 expression level was reduced in laryngeal carcinoma tissues and cell lines. Moreover, miR-379 overexpression inhibited the cell proliferation, migration, and invasion of laryngeal carcinoma cells. TCF-4 was detected as a direct target of miR-379 in laryngeal carcinoma. Furthermore, restored TCF-4 expression rescued the inhibitory roles of miR-379 overexpression on cell proliferation, migration, and invasion.Our study for the first time demonstrated that miR-379/TCF-4 might involve in the progression of laryngeal carcinoma, and miR-379 appears to serve as a novel tumor suppressor in laryngeal carcinoma.

Topics: Blotting, Western; Cell Line; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Neoplasms; Lipids; MicroRNAs; Neoplasm Invasiveness; Transcription Factor 7-Like 2 Protein

To investigate the effects of caveolin-1 on the biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line in vitro.. Eukaryotic expression vector of human caveolin-1 gene was constructed and transfected into HEp2 cells by Lipofectamine. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western blotting. Cell proliferation viability was tested by MTT assay. Anchorage-independent growth was determined by assaying colony formation in soft agar. Flow cytometry was used to assess the cell cycle and apoptosis. The relative phosphorylation level of EGFR and ERK1/2 were detected by Western blotting. Localization of caveolin-1 and EGFR were studied by laser confocal laser scanning microscopy.. The expression vector of caveolin-1 was constructed and three clones stably overexpressing caveolin-1 were obtained. Comparing with the parental HEp2 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of FACS analysis revealed that overexpression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction. EGFR was found to colocalize with caveolin-1 in transfected cells by confocal laser scanning microscopy and Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR and Erkl/2.. Overexpression of caveolin-1 suppresses the growth of HEp2 cells and induces apoptosis and inhibition of EGFR-MAPK signaling pathway may be involved in its mechanism.

Topics: Apoptosis; Blotting, Western; Carcinoma, Squamous Cell; Caveolin 1; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; ErbB Receptors; Flow Cytometry; Genetic Vectors; Humans; Laryngeal Neoplasms; Lipids; Microscopy, Confocal; Mitogen-Activated Protein Kinase 3; Phosphorylation; Polymerase Chain Reaction; Signal Transduction; Transfection