EB病毒BARF1原型和变异型基因对鼻咽癌细胞生物学行为的影响
发布时间:2019-05-17 15:08
【摘要】:背景和目的:EB病毒(Epstein-Barr virus, EBV)编码早期基因BARF1是新近确立的EBV致癌基因之一,其转化作用已在多种细胞系中得到证实。由于BARF1在多数鼻咽癌(nasopharyngeal carcinoma, NPC)和EBV相关胃癌组织中表达,推测其可能在NPC、胃癌等上皮细胞性肿瘤的发生中发挥重要作用。我们前期在检测BARF1基因多态性时发现V29A变异型在NPC中的检出率高于健康人群。本研究旨在探讨BARF1基因对NPC细胞系肿瘤原性的影响以及BARF1基因变异型对其致癌潜能的影响,为研究BARF1参与NPC的作用及机制提供依据。 方法:构建BARF1基因原型(B95-8)和变异型(V29A)慢病毒表达载体,分别转染EBV阴性NPC细胞系CNE,杀稻瘟菌素筛选,RT-PCR和qRT-PCR鉴定目的基因表达获得BARF1原型和变异型基因稳定转染细胞。以对照病毒转染细胞为对照,进行MTT实验、细胞周期检测、平板克隆形成实验、凋亡诱导实验和细胞迁移实验,检测BARF1原型和变异型基因对CNE细胞生物学行为的影响。 结果:(1)成功构建BARF1基因原型和变异型基因稳定转染细胞,2种基因在EBV阴性NPC细胞系中均能有效表达。(2)细胞增殖实验:BARF1原型和变异型基因转染细胞及对照病毒转染细胞MTT吸光度值、平板克隆形成率和细胞周期细胞增殖指数均无显著性差异(P均大于0.05)。(3)凋亡诱导实验:顺铂(2μg/ml)诱导48h,感染BARF1原型和变异型慢病毒细胞的存活率均高于对照病毒感染细胞,差异有显著性(P均小于0.05),而细胞凋亡率低于对照病毒感染细胞,差异有显著性(P均小于0.05),变异型与原型之间细胞存活率和凋亡率均无显著性差异(P均大于0.05)。(4)细胞迁移实验:BARF1原型和变异型慢病毒感染细胞的迁移能力高于对照病毒感染细胞,差异有显著性(P0.05),变异型与原型之间细胞迁移无显著性差异(P0.05)。 结论:(1)BARF1基因对EBV阴性NPC细胞系CNE无明显促增殖作用,但可提高其抗凋亡能力和迁移能力,从而增强CNE细胞肿瘤原性,在NPC发生发展过程中可能起重要作用。(2)BARF1V29A变异型对BARF1致癌潜能无明显影响。
[Abstract]:Background & AIM: EB virus (Epstein-Barr virus, EBV) coding early gene BARF1 is one of the newly established EBV carcinogenic genes, and its transformation has been confirmed in many cell lines. Because BARF1 is expressed in most nasopharyngeal carcinoma (nasopharyngeal carcinoma, NPC) and EBV related gastric cancer tissues, it is speculated that BARF1 may play an important role in the occurrence of NPC, gastric cancer and other epithelial cell tumors. When we detected the polymorphism of BARF1 gene, we found that the detection rate of V29A variant in NPC was higher than that in healthy population. The purpose of this study was to investigate the effect of BARF1 gene on tumorigenicity of NPC cell line and the effect of BARF1 gene variant on its carcinogenic potential, and to provide evidence for the study of the role and mechanism of BARF1 in NPC. Methods: BARF1 gene prototype (B95 鈮,
本文编号:2479195
[Abstract]:Background & AIM: EB virus (Epstein-Barr virus, EBV) coding early gene BARF1 is one of the newly established EBV carcinogenic genes, and its transformation has been confirmed in many cell lines. Because BARF1 is expressed in most nasopharyngeal carcinoma (nasopharyngeal carcinoma, NPC) and EBV related gastric cancer tissues, it is speculated that BARF1 may play an important role in the occurrence of NPC, gastric cancer and other epithelial cell tumors. When we detected the polymorphism of BARF1 gene, we found that the detection rate of V29A variant in NPC was higher than that in healthy population. The purpose of this study was to investigate the effect of BARF1 gene on tumorigenicity of NPC cell line and the effect of BARF1 gene variant on its carcinogenic potential, and to provide evidence for the study of the role and mechanism of BARF1 in NPC. Methods: BARF1 gene prototype (B95 鈮,
本文编号:2479195
本文链接:https://www.wllwen.com/yixuelunwen/yank/2479195.html
最近更新
教材专著