ERK Activation Modulates Cancer Stemness and Motility of a Novel Mouse Oral Squamous Cell Carcinoma Cell Line

Abstract

We established the NHRI-HN1 cell line from a mouse tongue tumor induced by 4-nitroquinoline 1-oxide (4-NQO)/arecoline, with further selection for cell stemness via in vitro sphere culture, to evaluate potential immunotherapies for oral squamous cell carcinoma (OSCC) in East and Southeast Asia. In vivo and in vitro phenotypic characterization, including tumor growth, immune modulator administration, gene expression, morphology, migration, invasion, and sphere formation assays, were conducted. NHRI-HN1 cells are capable of generating orthotopic tumors in syngeneic mice. Interestingly, immune stimulation via CpG oligodeoxynucleotide (CpG-ODN) dramatically reduced the tumor growth in NHRI-HN1 cell-injected syngeneic mice. The pathways enriched in genes that were differentially expressed in NHRI-HN1 cells when compared to non-tumorigenic cells were similar to those that were identified when comparing human OSCC and non-tumorous tissues. NHRI-HN1 cells have characteristics of epithelial–mesenchymal transition (EMT), including enhanced migration and invasion. NHRI-HN1 cells showed aggressive cell growth and sphere formation. The blockage of extracellular signal-regulated kinase (ERK) activation suppressed cell migration and reduced stemness characteristics in NHRI-HN1 cells, similar to human OSCC cell lines. Our data suggest that NHRI-HN1 cells, showing tumorigenic characteristics of EMT, cancer stemness, and ERK activation, are sufficient in modeling human OSCC and also competent for use in investigating oral cancer immunotherapies.

Establishment of a p53 Null Murine Oral Carcinoma Cell Line and the Identification of Genetic Alterations Associated with This Carcinoma

“…This is the only cell line that has been developed from a spontaneous tumor without exposure to any carcinogen. Normally, murine OSCC cell lines are either p53 mutant cell lines or p53 wild type cell lines [ 2 , 7 , 8 , 9 ]. However, MPC-1 is a p53 null cell line.…”

“…Although studies on human OSCC cell lines have allowed there to be remarkable research progress and an identification of new interventions, the immune-compromised preclinical platforms developed from these tumorigenic cell lines are not suitable for addressing questions related to the immune responses that target tumor growth [ 5 , 6 ]. Therefore, murine OSCC cell lines have been developed in the past few years in order to generate immune-competent preclinical models that can be used to assess tumor targeting [ 2 , 7 , 8 , 9 ].…”

“…It has been widely validated as an inducer of oral carcinogenesis in rodent models [ 6 , 10 , 11 ]. More than ten 4NQO-based murine tongue SCC cell lines, including six from our laboratory, have been generated [ 2 , 7 , 8 , 9 ]. Chen et al [ 9 ] established the NHRI-HN cell line series from the murine tongue tumors induced by 4NQO and arecoline.…”

“…More than ten 4NQO-based murine tongue SCC cell lines, including six from our laboratory, have been generated [ 2 , 7 , 8 , 9 ]. Chen et al [ 9 ] established the NHRI-HN cell line series from the murine tongue tumors induced by 4NQO and arecoline. The syngeneic growth of the NHRI-HN1 cell line was found to be suppressed by an immune stimulant.…”

Inhibitory role of ATF3 in gastric cancer progression through regulating cell EMT and stemness

“…The study by Srivastava et al [ 10 ] showed FAT1 gene, an ortholog of Drosophila tumor suppressor gene fat, modulates EMT and stemness genes expression in hypoxic glioblastoma. ERK was also found to suppress oral squamous carcinoma cell migration and reduce stemness characteristics [ 11 ]. Although hyperproliferative capacity and overproduction of gastric tumor cells are likely closely related to EMT and tumor stemness [ 12 , 13 ], data about identification of molecular markers pertinent to pathogenic process and potential therapy are largely lacking.…”