Abstract
ObjectiveTo investigate the effects of NK4 gene on the properties and tumorigenicity in laryngeal squamous cell carcinoma cell.MethodsHere, we used the attenuated Salmonella carrying the NK4 gene to transfect the AMC-HN-8 cells and detected the expression of NK4 by the real-time quantitative polymerase chain reaction (q RT-PCR). The properties of NK4 gene was determined by MTT method, cell scratch test, and flow cytometry. A nude mouse tumorigenesis model was used to evaluate the effect of NK4 gene on the growth of AMC-HN-8 cells in vivo. While a western blot assay was used to assess the expression of DKK1, Wnt1 and β-Catenin in nude mouse tumors.ResultsqRT-PCR showed that the expression of NK4 in the transfection group was significantly higher than that in the control group (P<0.01), and the expression increased with the time of transfection. MTT results showed NK4 overexpression inhibited the proliferation of AMC-HN-8 cells, and the inhibitory activity no longer increased with increasing dose when 30% expression supernatant was added (P<0.01). Scratch experiment showed that NK4 overexpression decreased the cell migration ability (P<0.01). Annexin V/PI double staining experiment showed that NK4 gene induced AMC-HN-8 cell apoptosis (P<0.01), and cell cycle arrest in S phase (P<0.01). NK4 overexpression inhibited tumor formation ability of AMC-HN-8 cells in vivo (P <0.05). WB detection showed that the expression of DKK1 increased, Wnt1 and β-Catenin protein decreased after the high expression of NK4.ConclusionsNK4 gene inhibit cell proliferation and migration, while promote cell apoptosis, and induce cell cycle arrest in S phase of laryngeal carcinoma AMC-HN-8 cells. NK4 overexpression inhibit the tumorigenesis ability of AMC-HN-8 cells, which may be related to the regulation of DKK1/Wnt1/β-Catenin signal axis.
Highlights
Laryngeal cancer is the most common malignancy in the head and neck, accounting for 1-5% of systemic malignancies, of which 96-98% are laryngeal squamous cell carcinoma (LSCC)
Based on the dual antitumor properties of the NK4 gene and the high expression of Wnt1/b-Catenin signaling pathway in the head and neck, this study aims to explore the following issues: [1] NK4 expression in transfected LSCC AMC-HN-8 cells; [2] the effects of the NK4 gene on the proliferation, migration, apoptosis and cell cycle of LSCC AMC-HN-8 cells;(3) The effect of NK4 gene on the tumorigenesis ability of nude mice of laryngeal cancer cells, and to explore whether its DKK1/Wnt1/b-Catenin signaling pathway is involved in the effect of NK4 gene on laryngeal cancer
NK4 acts like an antagonist, inhibiting Hepatocyte growth factor (HGF)/c-Met induced tumor growth, metastasis and invasion, and inhibits vascular endothelial growth factor (VEGF) - and basic fibroblast growth factor-induced tumor angiogenesis, which is independent of the HGF/c-Met pathway, and causes the tumor cell apoptosis
Summary
Laryngeal cancer is the most common malignancy in the head and neck, accounting for 1-5% of systemic malignancies, of which 96-98% are laryngeal squamous cell carcinoma (LSCC). A study in 2018 [1] estimated that 177,422 new cases of laryngeal cancer and 94,771 deaths. Surgery is still the most effective treatment. Chemotherapy and novel gene-targeted therapy have led to significant progress in the treatment of laryngeal cancer. Gene-targeted therapy has shown good development and application prospects in cancer treatment. Seeking new approaches to treat laryngeal cancer at the gene level has become a new challenge faced by otolaryngologists
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
