人胃上皮细胞中表达的 Epstein-Barr 病毒编码的 BARF1 基因的作用。

The role of the Epstein-Barr virus-encoded BARF1 gene expressed in human gastric epithelial cells.

机构信息

North China University of Science and Technology (Hebei Key Laboratory for Chronic Diseases, Tangshan Key Laboratory for Preclinical and Basic Research on Chronic Diseases), Hebei Province, P.R. China.

College of Life Science and Bioengineering, Beijing University of Technology, Beijing, P.R. China.

出版信息

Turk J Gastroenterol. 2020 Nov;31(11):775-781. doi: 10.5152/tjg.2020.18827.

DOI:10.5152/tjg.2020.18827

PMID:33361040

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7759223/

Abstract

BACKGROUND/AIMS: The study aimed to explore the effects of Epstein-Barr virus--encoded BARF1 in human gastric epithelial cells (GES-1).

MATERIALS AND METHODS

A eukaryotic expression vector carrying BARF1 gene (pcDNA3.1-BARF1) was constructed. The pcDNA3.1-BARF1 was transfected into GES-1 cells, and they were selected by G418. The GES-1 cells lines that expressed BARF1 (GES-1-BARF1) were obtained. The cycle of GES-1-pcDNA3.1 cells (GES-1 cells transfected with empty vector), GES-1-BARF1 cells (GES-1 cells transfected with BARF1), and TPA-GES-1-BARF1(GES-1-BARF1 cells stimulated by 12-O-tetradecanoylphorbol-13-acetate (TPA) were analyzed by flow cytometry. Colony formation in soft agar and tumorigenicity of the transfected cells in mice with severe combined immunodeficiency (SCID) were also observed.

RESULTS

The morphology of GES-1-BARF1 cells were changed from the original shuttle to round, the adhesion between the cells and bottle wall was weakened, and the cells showed overlapping growth. The proliferation rate of GES-1-BARF1 and TPA-GES-1-BARF1 cells were faster than GES-1 and GES-1-pcDNA3.1 cells; the S phase was significantly prolonged for GES-1-BARF1 and TPA-GES-1-BARF1. GES-1-BARF1 and TPA-GES-1-BARF1 cells formed colonies in soft agar, with a cloning rate of 24.2% (58/240) and 40.0% (96/240), respectively; GES-1 and GES-1-pcDNA3.1 cells did not form colonies in soft agar. Tumors were formed in mice with SCID after injecting TPA-GES-1-BARF1 cell groups. Tumor formation did not occur in mice with SCID after injecting GES-1 and GES-1-pcDNA3.1 cell groups, but nodules were formed in the mice with SCID after injecting GES-1-BARF1 cell groups.

CONCLUSION

GES-1-BARF1 cells malignant transformation was induced by transfected BARF1 gene and TPA stimulation. This result indicated that tumor formation not only require oncogenes, but also the stimulation of cancer-promoting substance.

摘要

背景/目的：本研究旨在探讨 Epstein-Barr 病毒编码的 BARF1 在人胃上皮细胞（GES-1）中的作用。

材料和方法

构建携带 BARF1 基因的真核表达载体（pcDNA3.1-BARF1）。将 pcDNA3.1-BARF1 转染至 GES-1 细胞，并用 G418 进行筛选。获得表达 BARF1 的 GES-1 细胞系（GES-1-BARF1）。通过流式细胞术分析 GES-1-pcDNA3.1 细胞（转染空载体的 GES-1 细胞）、GES-1-BARF1 细胞（转染 BARF1 的 GES-1 细胞）和 TPA-GES-1-BARF1 细胞（用 12-O-十四烷酰佛波醇-13-乙酸酯（TPA）刺激的 GES-1-BARF1 细胞）的细胞周期。还观察了转染细胞在严重联合免疫缺陷（SCID）小鼠中的软琼脂集落形成和致瘤性。

结果

GES-1-BARF1 细胞的形态由原来的梭形变为圆形，细胞与瓶壁的黏附力减弱，细胞呈重叠生长。GES-1-BARF1 和 TPA-GES-1-BARF1 细胞的增殖速度快于 GES-1 和 GES-1-pcDNA3.1 细胞；GES-1-BARF1 和 TPA-GES-1-BARF1 的 S 期明显延长。GES-1-BARF1 和 TPA-GES-1-BARF1 细胞在软琼脂中形成集落，克隆率分别为 24.2%（58/240）和 40.0%（96/240）；GES-1 和 GES-1-pcDNA3.1 细胞在软琼脂中未形成集落。在注射 TPA-GES-1-BARF1 细胞组的 SCID 小鼠中形成肿瘤。在注射 GES-1 和 GES-1-pcDNA3.1 细胞组的 SCID 小鼠中未形成肿瘤，但在注射 GES-1-BARF1 细胞组的 SCID 小鼠中形成了结节。