Development and applications of a nasopharyngeal carcinoma Tet-Off cell line

Feng,Yu-Peng; Liu,Qi-Cai; Zhu,Jun-Feng; Xie,Fu-Kang; Li,Li

doi:10.3892/ol.2011.262

Development and applications of a nasopharyngeal carcinoma Tet-Off cell line

Authors:
- Yu-Peng Feng
- Qi-Cai Liu
- Jun-Feng Zhu
- Fu-Kang Xie
- Li Li
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Affiliations: Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, P.R. China, Experimental Medical Research Center, Guangzhou Medical College, Guangzhou 510182, P.R. China, Department of Pathology, The First Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510060, P.R. China, Imaging Diagnosis and Interventional Center, Cancer Center, Sun Yat-Sen University, Guangzhou 510060, P.R. China
Published online on: March 1, 2011 https://doi.org/10.3892/ol.2011.262
Pages: 525-530

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Abstract

The conditional activation and inactivation of target gene expression in a nasopharyngeal carcinoma (NPC) cell line is beneficial for the study of the roles of NPC-related genes. Based on the Tet-Off Advanced system, a NPC S18 Tet-Off cell line was developed by stable transfection of a pTet-Off Advanced vector (regulator plasmid in Tet-Off Advanced system) into NPC S18 cells. Doxycycline-dependent regulators expressed in the S18 Tet-Off cells were examined by transient and stable transfection of pTRE-Tight-Luc. The S18 Tet-Off-Luc clone selected by stable transfection of pTRE-Tight-Luc into S18 Tet-Off cells expressed firefly luciferase under tight control of doxycycline in a time- and dose-dependent manner. To test applications of the S18 Tet-Off cell line in the study of gene function, the impact of ferritin heavy chain (FTH1) gene on NPC cell growth was examined. The S18 Tet-Off-FTH1 clone was developed by stably transfecting pTRE-Tight-FTH1 (response plasmid harboring FTH1) into S18 Tet-Off cells. FTH1 levels in the S18 Tet-Off-FTH1 clone were semi-quantitatively regulated in response to varying concentrations of doxycycline. A cell proliferation assay showed that a high expression of FTH1 (cells grown in the absence of doxycycline) reduced cell growth, while moderate FTH1 overexpression (cells grown in 0.1 ng/ml doxycycline) had no adverse effect on cell growth. In conclusion, the S18 Tet-Off cell line provides a proven genetic background for convenient access to controllable gene expression in NPC.

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