CNE2 stem-like cells of nasopharyngeal carcinoma have stem cell characteristics and high level of autophagy

doi:10.3969/j.issn.2095-4344.1718

Abstract

Abstract:

BACKGROUND: Nasopharyngeal carcinoma is a head and neck malignant tumor with high invasion and metastatic potential. It has not yet achieved satisfactory effects in the treatment of advanced nasopharyngeal carcinoma. Some autophagy inhibitors and anti-tumor drugs have been used in clinical research, but the possible mechanism has not been clarified.
OBJECTIVE: To explore whether high-level autophagy occurs in stem-like cells of nasopharyngeal carcinoma.
METHODS: Tumor stem-like cell spheres were enriched from humanized nasopharyngeal carcinoma CNE2 cell line by serum-free suspension culture. The stem cell characteristics and expression levels of autophagy-related genes and proteins in CNE2 parental cells and CNE2 stem-like cells were detected using cell differentiation assay, flow cytometry, transmission electron microscope, western blot and RT-PCR.
RESULTS AND CONCLUSION: The nasopharyngeal carcinoma CNE2 stem-like cells were enriched by serum-free suspension culture. The suspension spheres were cultured in serum-containing medium to differentiate into CNE2 parental cells. Proportion of CD133⁺ cells in CNE2 stem-like cells (9.6%) was significantly higher than that in CNE2 parental cells (0.3%; P < 0.01). Compared with CNE2 parental cells, CNE2 stem-like cells highly expressed stemness-related genes Bmi-1, Twist1 and autophagy-related genes Beclin1 and LC3B (P < 0.01). Under the transmission electron microscope, the autophagic bodies of CNE2 stem-like cells in nasopharyngeal carcinoma were significantly increased compared with CNE2 parental cells. Therefore, nasopharyngeal carcinoma CNE2 stem-like cells can be effectively enriched using the serum-free suspension culture method, and be verified to have the biological characteristics of stem cells and exhibit high-level autophagy.

Key words: nasopharyngeal carcinoma, tumor stem cells, serum-free suspension culture, nasopharyngeal carcinoma stem-like cells, autophagy, National Natural Science Foundation of China

CLC Number:

Dai Senlin, Li Xiaoqian, Wu Jie, Yue Changwu, Li Yajun. CNE2 stem-like cells of nasopharyngeal carcinoma have stem cell characteristics and high level of autophagy[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(17): 2703-2708.

Figures/Tables 2

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