人乳头瘤病毒16型阳性扁桃体癌在BALB/c裸鼠体内异种移植瘤的建立与鉴定

Establishment and characterization of a human papillomavirus type 16-positive tonsillar carcinoma xenograft in BALB/c nude mice.

作者信息

Letsolo Boitelo T, Faust Helena, Ekblad Lars, Wennerberg Johan, Forslund Ola

机构信息

Department of Laboratory Medicine, Division of Medical Microbiology, Lund University and Skåne Regional and University Laboratories, Malmö, Sweden.

School of Molecular and Cell Biology, University of the Witwatersrand, Johannesburg, Republic of South Africa.

出版信息

Head Neck. 2016 Mar;38(3):417-25. doi: 10.1002/hed.23918. Epub 2015 Jul 15.

DOI:10.1002/hed.23918

PMID:25352201

Abstract

BACKGROUND

Among head and neck cancers, human papillomavirus type 16 (HPV16) is associated with tonsillar carcinomas. Despite this, no HPV16-positive tonsillar cancer cell line has been established in nude mice.

METHODS

Fresh tonsillar carcinoma biopsies were obtained from 23 patients and implanted subcutaneously into nude mice (BALB/c, nu/nu).

RESULTS

After 7 months, one xenograft was established. The primary tumor harbored 2.7 copies (95% confidence interval = 2.4-2.9) of HPV16/cell and displayed 99.9% (7904/7906) nucleotide identity to HPV16 (EU118173.1). The xenograft showed increased methylation in two E2-binding sites of the HPV16 genome. Both episomal and integrated HPV16 were detected in the original tumor and in 14 xenografts from the second passage. From this passage, a viral load of 6.4 copies/cell (range = 4.6-9.6) and 3.7 (range = 1.0-5.5) E7-mRNA transcripts/HPV16-genome were detected.

CONCLUSION

This xenograft represents the first established HPV16-positive tonsillar tumor in nude mice and could provide an experimental system of HPV16-positive tonsillar cancers.

摘要

背景

在头颈癌中，16型人乳头瘤病毒（HPV16）与扁桃体癌相关。尽管如此，尚未在裸鼠中建立HPV16阳性的扁桃体癌细胞系。

方法

从23例患者获取新鲜扁桃体癌活检组织，并皮下植入裸鼠（BALB/c，nu/nu）。

结果

7个月后，建立了一个异种移植瘤。原发肿瘤中HPV16的拷贝数为2.7（95%置信区间=2.4-2.9）/细胞，与HPV16（EU118173.1）的核苷酸同一性为99.9%（7904/7906）。该异种移植瘤在HPV16基因组的两个E2结合位点显示甲基化增加。在原发肿瘤以及第二代传代的14个异种移植瘤中均检测到游离型和整合型HPV16。从这一代开始，检测到病毒载量为6.4拷贝/细胞（范围=4.6-9.6），E7-mRNA转录本为3.7（范围=1.0-5.5）/HPV16基因组。