LTF基因在鼻咽癌细胞系中生物学功能的初步研究

目的:初步探讨野生型LTF基因在鼻咽癌细胞系中的生物学功能。方法:野生型LTF导入鼻咽癌细胞系,G418筛选,RT-PCR和Western-blotting分别在mRNA和蛋白质水平进行验证,得到稳定表达LTF基因的鼻咽癌细胞系。流式细胞术、平板克隆形成实验和MTT法分别检测细胞周期、细胞的克隆形成能力和细胞生长曲线。结果:成功导入LTF并稳定表达的鼻咽癌细胞系,G0-G1期细胞百分比例明显增加(72.01%vs 62.31%),G2-M期细胞百分比例减少(6.26%vs 10.81%);克隆形成能力降低(39.5%vs 59.7%),体外瘤细胞增殖能力降低(P0.05)。结论:LTF基因可阻滞细胞周期、抑制鼻咽癌细胞系的增殖能力和克隆形成率,同时为进一步的体内试验研究奠定基础。

The Preliminary Study of Biological Activity of LTF Gene in NPCDerived Cell Line

Objective:To preliminarily study the biological characteristics of lactotransferrin (LTF) gene in nasopharyngeal cancinoma(NPC) derived cell lines.Methods:Wild-typed LTF gene was transfected into NPC derived cell line. After G418 selection and confirmationthe expression at mRNA and protein level by RT-PCR and Western blotting analysis respectively, Cell cycle, cell proliferationand colony formation ability were detected by flow cytometry(FCM) analysis, MTT analysis and colony formation assay respectively. Results:LTF expression in NPC cell line could block the cell cycle progression in G1 phase, leading to an increase in the number of cells inG1 phase ( 72.01 %vs 62.31 %) and a decrease in the number of cells in G2/Mphase (6.26%vs 10.81%); And they had a much lowercolony efficiency ( 39.5%vs 59.7 %) and proliferated much more slowly(P < 0.05).Conclusion:Evidence showed that LTF blocked thecell cycle progression in G1 phase and had an inhibitory effect on proliferation and colony formation efficiency of NPC cells. The obtainmentpaved the way for further study on biological functions of LTF in vivo.