Identification and characterization of a highly metastatic epithelial cancer cell line from rat tongue cancer

Xing Qin; Ming Yan; Rongrong Li; Dongxia Ye; Jianjun Zhang; Qin Xu; Yuanyong Feng; Qiang Sun; Canhua Jiang; Wantao Chen

doi:10.1016/j.archoralbio.2018.07.010

Identification and characterization of a highly metastatic epithelial cancer cell line from rat tongue cancer

Arch Oral Biol. 2018 Nov:95:58-67. doi: 10.1016/j.archoralbio.2018.07.010. Epub 2018 Jul 17.

Authors

Xing Qin¹, Ming Yan², Rongrong Li³, Dongxia Ye⁴, Jianjun Zhang⁵, Qin Xu⁶, Yuanyong Feng⁷, Qiang Sun⁸, Canhua Jiang⁹, Wantao Chen¹⁰

Affiliations

¹ Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: xingzaing@163.com.
² Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: yanming8012@126.com.
³ Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: rongrli2003@163.com.
⁴ Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: yedongxia122@hotmail.com.
⁵ Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: zjjshuobo@163.com.
⁶ Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: xuqin_2004@hotmail.com.
⁷ Department of Stomatology, Affiliated Hospital of Qingdao University, Qingdao, 266071, China. Electronic address: walterf@163.com.
⁸ Department of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China. Electronic address: sunqiang850128@hotmail.com.
⁹ Department of Oral and Maxillofacial Surgery, Xiangya Hospital of Central South University, 87 Xiangya Road, Changsha, Hunan, 410008, China. Electronic address: canhua-j@sohu.com.
¹⁰ Department of Oral and Maxillofacial-Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, National Clinical Research Center of Stomatology, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. Electronic address: chenwantao196323@sjtu.edu.cn.

PMID: 30059808
DOI: 10.1016/j.archoralbio.2018.07.010

Abstract

Objectives: Tongue squamous cell carcinoma (TSCC) is a clinically devastating disease. However, most established TSCC cell lines currently show undesirable malignant behaviours. The purpose of this study is to establish a highly metastatic TSCC cell line to serve as a useful tool for basic research.

Materials and methods: TSCCs were induced by 4-nitroquinoline-1-oxide (4NQO) in Sprague-Dawley rats. Tumor cells were obtained from the cancer tissues by primary culture and were then purified by an in vitro invasion assay and a limiting dilution assay. The growth rate, cell cycle distribution, apoptotic rate, tumorigenicity and distant metastatic phenotypes of the rat tongue cancer cells were fully investigated and characterized.

Results: To date, the rat tongue cancer cell line, named Rca-T, has been continuously cultured in vitro for over 210 passages and exhibit a long spindle-shaped morphology, adherent growth, and a stable epithelial phenotype. The population doubling time of Rca-T cells is 23.35 h. Approximately 39.8% of these cells are in S phase, and the apoptosis rate of Rca-T cells is 7.46%. Furthermore, in immunodeficient nude mice, both the xenograft rate and the incidence of experimental lung metastasis are 100%. The in vitro assays further reveal the highly malignant and epithelial-mesenchymal transition-like properties of Rca-T cells.

Conclusion: In this study, the tumorigenic and highly distant metastatic TSCC cell line Rca-T was established. The malignant features of this cell line, especially its metastatic potential, will enable a wealth of functional studies on the molecular mechanisms of TSCC metastasis in the future.

Keywords: 4NQO; Cancer cell line; Highly metastatic; Oral squamous cell carcinoma; Rat tongue cancer.

MeSH terms

Animals
Apoptosis
Carcinogenesis
Carcinoma, Squamous Cell / pathology*
Cell Cycle
Cell Line, Tumor
Disease Models, Animal
Epithelial-Mesenchymal Transition
Lung Neoplasms / secondary
Mice, Nude
Neoplasm Metastasis / pathology*
Phenotype
Rats
Rats, Sprague-Dawley
Tongue Neoplasms / pathology*
Tumor Cells, Cultured