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

Kuo-Wei Chang,Hsi-Feng Tu,Hsin-Yao Chung,Yi-Fen Chen,Chia-En Lin,Shu-Chun Lin

doi:10.3390/ijms21249354

Abstract

Head and neck squamous cell carcinoma (HNSCC), including oral squamous cell carcinoma (OSCC), ranks sixth in cancer incidence worldwide. To generate OSCC cells lines from human or murine tumors, greatly facilitates investigations into OSCC. This study describes the establishing of a mouse palatal carcinoma cell line (designated MPC-1) from a spontaneous tumor present in a heterozygous p53 gene loss C57BL/6 mouse. A MPC-1-GFP cell subclone was then generated by lentivirus infection resulting in stable expression of green fluorescent protein. Assays indicated that MPC-1 was a p53 null polygonal cell that was positive for keratinocyte markers; it also expressed vimentin and showed a loss of E-cadherin expression. Despite that MPC-1 having strong proliferation and colony formation capabilities, the potential for anchorage independent growth and tumorigenesis was almost absent. Like other murine MOC-L and MTCQ cell line series we have previously established, MPC-1 also expresses a range of stemness markers, various oncogenic proteins, and a number of immune checkpoint proteins at high levels. However, the synergistic effects of the CDK4/6 inhibitor palbociclib on other therapeutic drugs were not observed with MPC-1. Whole exon sequencing revealed that there were high rates of non-synonymous mutations in MPC-1 affecting various genes, including Akap9, Arap2, Cdh11, Hjurp, Mroh2a, Muc4, Muc6, Sp110, and Sp140, which are similar to that the mutations present in a panel of chemical carcinogenesis-related murine tongue carcinoma cell lines. Analysis has highlighted the dis-regulation of Akap9, Cdh11, Muc4, Sp110, and Sp140 in human HNSCC as indicated by the TCGA and GEO OSCC databases. Sp140 expression has also been associated with patient survival. This study describes the establishment and characterization of the MPC-1 cell line and this new cell model should help to advance genetic research into oral cancer.

Highlights

Head and neck squamous cell carcinomas (HNSCCs), which include oral SCC (OSCC), are prevalent malignancies around the world [1,2]
This study describes the establishing of a new murine OSCC cell line
Most of the previously established cell lines are carcinogen-associated tumor cell lines, many created using 4-nitroquinoline 1-oxide (4NQO) exposure. This is the only cell line that has been developed from a spontaneous tumor without exposure to any carcinogen

Summary

Introduction

Head and neck squamous cell carcinomas (HNSCCs), which include oral SCC (OSCC), are prevalent malignancies around the world [1,2]. 4-nitroquinoline 1-oxide (4NQO) is a water-soluble carcinogen that possesses tobacco-mimicking effects; it induces a range of oxidative species and DNA adducts, as well as acts as a mutagen [10] It has been widely validated as an inducer of oral carcinogenesis in rodent models [6,10,11]. Chen et al [9] established the NHRI-HN cell line series from the murine tongue tumors induced by 4NQO and arecoline. Wang et al [2] generated the 4MOSC cell line series from the 4NQO-induced tongue SCC in mice. The MOC-L series of OSCC cell lines were established from 4NQO-induced tumors induced in these mice [8]. We established the MTCQ cell line series from 4NQO-induced tongue SCC [7].

Methods

Results

Conclusion