NK4 Regulates Laryngeal Squamous Cell Carcinoma Cell Properties and Inhibits Tumorigenicity by Modulating the DKK1/Wnt/β-Catenin Axis

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.

Dickkopf‐1 drives perineural invasion via PI3K–AKT signaling pathway in head and neck squamous cancer

“…27 The DKK1-Wntβ-catenin signaling axis is negatively regulated by the NK4 gene in HNSCC, thus mediating the occurrence and development of HNSCC. 28 DKK1 can also activate the Wntplanar cell polarity pathway to promote neuronal death in Alzheimer's disease. 29 However, the mechanism of DKK1 and PNI remains unknown.…”

“…Upregulated expression of DKK1 in colorectal cancer can inhibit CD8 + T cells through the GSK3β–E2F1–T‐BET pathway and promote malignant progression of cancer cells 27 . The DKK1–Wnt–β‐catenin signaling axis is negatively regulated by the NK4 gene in HNSCC, thus mediating the occurrence and development of HNSCC 28 . DKK1 can also activate the Wnt‐planar cell polarity pathway to promote neuronal death in Alzheimer's disease 29 .…”

Identification of an immune-related genes signature to predict risk of recurrence for patients with laryngeal squamous cell carcinoma

“…Researches have revealed that DDK1 is over-expressed in LSCC tissues and can be served as an unfavorable survival predictor, which may be a promising therapeutic target for LSCC patients. 38 , 39 Another IRG, namely, interleukin 1 receptor accessory protein (IL1RAP), acts as an IL1R co-receptor to mediate the activation of IL-1 responsive genes. 40 In recent years, many researchers have found that IL1RAP as a key tumor-associated antigen might be used as an immunotherapy target.…”