Tong Dongdong, Sun Jing, Huang Ping, Li Minqi, Zhang Fenghe
Department of Oral and Maxillofacial Surgery, School of Stomatology, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Shandong University, Jinan, Shandong 250012, P.R. China.
Department of Bone Metabolism, School of Stomatology, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Shandong University, Jinan, Shandong 250012, P.R. China.
Mol Med Rep. 2017 May;15(5):2521-2529. doi: 10.3892/mmr.2017.6291. Epub 2017 Mar 7.
The present study detected p75 neurotrophin receptor (p75NTR) expression in tongue squamous cell carcinoma (TSCC) cell lines, in order to define the biological properties of p75NTR+ cells and to confirm the use of p75NTR+ as a surface marker for TSCC stem cells. p75NTR+ cells were separated from Tca‑8113 and CAL‑27 TSCC cells by fluorescence‑activated cell sorting. Colony formation, MTT and scratch assays, and a tumorigenicity analysis were performed to measure self-renewal and proliferation, multidirectional differentiation, and tumorigenicity of p75NTR+ cells. p75NTR+ cells comprised 3.1 and 1.9% of Tca‑8113 and CAL‑27 cells (mean of three experiments), respectively, and were more able to form colonies compared with non‑sorted cells (P<0.01). In addition, the proportion of p75NTR+ cells generated from monoclonal p75NTR+ cells decreased to 14.5 (Tca‑8113) and 5.8% (CAL‑27) of cells within 2 weeks, thus suggesting that p75NTR+ cells are able to generate p75NTR+ and p75NTR‑ cells. Furthermore, p75NTR+ cells exhibited increased proliferation, as evidenced by MTT assay (P<0.01) and had greater metastatic ability according to the scratch assay (P<0.01), compared with non‑sorted cells. p75NTR+ cells also exhibited a greater tumorigenic capacity compared with non‑sorted cells. In conclusion, p75NTR+ cells isolated from TSCC cell lines possess the characteristics of cancer stem cells; therefore, p75NTR may be considered a useful surface marker for the identification of TSCC stem cells.
本研究检测了舌鳞状细胞癌（TSCC）细胞系中p75神经营养因子受体（p75NTR）的表达，以确定p75NTR+细胞的生物学特性，并证实p75NTR+可作为TSCC干细胞的表面标志物。通过荧光激活细胞分选从Tca-8113和CAL-27 TSCC细胞中分离出p75NTR+细胞。进行集落形成、MTT和划痕试验以及致瘤性分析，以检测p75NTR+细胞的自我更新和增殖、多向分化及致瘤性。p75NTR+细胞分别占Tca-8113和CAL-27细胞的3.1%和1.9%（三个实验的平均值），与未分选细胞相比，其形成集落的能力更强（P<0.01）。此外，单克隆p75NTR+细胞产生的p75NTR+细胞比例在2周内降至细胞总数的14.5%（Tca-8113）和5.8%（CAL-27），这表明p75NTR+细胞能够产生p75NTR+和p75NTR-细胞。此外，MTT试验证明p75NTR+细胞的增殖能力增强（P<0.01），划痕试验显示其转移能力更强（P<0.01），与未分选细胞相比。p75NTR+细胞的致瘤能力也比未分选细胞更强。总之，从TSCC细胞系中分离出的p75NTR+细胞具有癌症干细胞的特征；因此，p75NTR可被视为鉴定TSCC干细胞的有用表面标志物。
