Abstract
To investigate effects of the TESTIN (TES) gene on proliferation and migration of highly metastatic nasopharyngeal carcinoma cell line 5-8F and the related mechanisms. The target gene of human nasopharyngeal carcinoma cell line 5-8F was amplified by PCR and cloned into the empty plasmid pEGFP-N1 to construct a eukaryotic expression vector pEGFP-N1-TES. This was then transfected into 5-8F cells. MTT assays, flow cytometry and scratch wound tests were used to detect the proliferation and migration of transfected 5-8F cells. A cell model with stable and high expression of TES gene was successfully established. MTT assays showed that the OD value of 5-8F/TES cells was markedly lower than that of 5-8F/GFP cells and 5-8F cells (p<0.05). Flow cytometry showed that the apoptosis rate of 5-8F/TES cells was prominently increased compared with 5-8F/GFP cells and 5-8F cells (p<0.05). In vitro scratch wound assays showed that, the width of the wound area of 5-8F/TES cells narrowed slightly, while the width of the wound area of 5-8F/ GFP cells and 5-8F cells narrowed sharply, suggesting that the TES overexpression could inhibit the migration ability. TES gene expression remarkably inhibits the proliferation of human nasopharyngeal carcinoma cell line 5-8F and reduces its migration in vitro. Thus, it may be a potential tumor suppressor gene for nasopharyngeal carcinoma.
Highlights
Nasopharyngeal carcinoma is one of the common clinical head and neck cancers with a relatively young age of onset
The recombinant plasmid pEGFP-N1-TES was identified by PCR and DNA sequencing
The TES gene is isolated from human nasopharyngeal carcinoma cells and inserted into PEGFP-N1 vector to construct a recombinant plasmid PEGFP-N1-TES in vitro
Summary
Nasopharyngeal carcinoma is one of the common clinical head and neck cancers with a relatively young age of onset. TES is widely expressed in normal tissues but deficit in a variety of primary tumors and tumor cell lines, especially in the cell lines of gastric carcinoma, breast cancer, colorectal cancer, glioma, ovarian cancer, leukemia and endometrial carcinoma, suggesting that it may be a candidate tumor suppressor gene (Zenklusen et al, 1995; Bieche et al, 1997; Edelson et al, 1997; Nishizuka et al, 1997; Weeks et al, 2010; Gu et al, 2014). This study was conducted to investigate the effects of TES gene overexpression on the proliferation and migration of human nasopharyngeal carcinoma cell 5-8F, and explore the related mechanism
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