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目的:探讨Testin(TES)基因对人鼻咽鳞状细胞癌株5-8F细胞增殖、迁移的影响。方法:以RTPCR法从人鼻咽鳞状细胞癌株5-8F细胞获得目的基因,构建真核表达载体pEGFP-N1-TES,采用PCR、基因测序等对其进行鉴定;然后将其转导入人高转移鼻咽鳞状细胞癌株5-8F中,通过RT-PCR法及Western-blot法对其进行鉴定。接着采用流式细胞术、划痕试验检测转染后5-8F细胞增殖及迁移能力的影响。结果:转染外源性TES基因后,转染外源性TES基因的5-8F细胞呈现明显的凋亡,差异有统计学意义(P<0.05)。细胞划痕实验表明:在第12、24、48h时,TES组细胞迁移速率明显比空载体组和未转染组要慢(P<0.01)。结论:该研究成功建立了能稳定高表达TES的细胞模型。TES基因在体外可明显抑制鼻咽癌5-8F细胞的增殖和降低其迁移运动能力,可能是一种潜在的抑癌基因。
Objective: To investigate the effect of Testin (TES) gene on the proliferation and migration of human nasopharyngeal squamous cell carcinoma cell line 5-8F. Methods: The target gene was obtained from human nasopharyngeal squamous cell carcinoma cell line 5-8F by RTPCR method. The eukaryotic expression vector pEGFP-N1-TES was constructed and identified by PCR and gene sequencing. Highly metastatic nasopharyngeal squamous cell carcinoma cell line 5-8F was identified by RT-PCR and Western-blot. Next, flow cytometry and scratch assay were used to detect the effect of 5-8F cells proliferation and migration after transfection. Results: After transfection of exogenous TES gene, 5-8F cells transfected with exogenous TES gene showed obvious apoptosis, the difference was statistically significant (P <0.05). Cell scratch assay showed that at the 12th, 24th and 48th hour, the cell migration rate in TES group was significantly slower than that in empty vector group and non-transfected group (P <0.01). Conclusion: This study successfully established a stable and high expression of TES cell model. TES gene in vitro can significantly inhibit 5-8F nasopharyngeal carcinoma cell proliferation and reduce their ability to migrate, may be a potential tumor suppressor gene.