Palanca-Wessels M C, Barrett M T, Galipeau P C, Rohrer K L, Reid B J, Rabinovitch P S
Department of Pathology, University of Washington, Seattle, USA.
Gastroenterology. 1998 Feb;114(2):295-304. doi: 10.1016/s0016-5085(98)70480-9.
BACKGROUND & AIMS: Progression to cancer in Barrett's esophagus occurs through an accumulation of cell cycle and genetic abnormalities that have been documented in vivo. To better study neoplastic evolution in Barrett's esophagus, the aim of this study was to establish in vitro cultures from preneoplastic tissues.
Mechanical and enzymatic dissociation methods were used to initiate Barrett's epithelial cultures from endoscopic biopsy specimens, and the cells were characterized using flow-cytometric, cytogenetic, and molecular genetic analyses.
Four long-term cultures were established from 39 attempts. All cultures contain cytogenetic abnormalities and elevated flow-cytometric 4N DNA content fractions. Molecular genetic abnormalities detected include the following: 9p and/or CDKN2/p16 abnormalities in 4 of 4 cultures, 17p loss of heterozygosity and p53 mutation in 3 of 4 cultures, and 5q loss of heterozygosity in 1 of 4 cultures. Inactivation of p53 was statistically associated with successful long-term culture.
These cultures contain cell cycle and molecular genetic abnormalities that closely parallel those previously documented to occur early in cancer development in Barrett's esophagus in vivo. These alterations also appear to be associated with successful growth in vitro. The cultures may provide a premalignant in vitro system in which to test potential therapies for Barrett's esophagus as well as to examine etiologic factors and genetic intermediates important in neoplastic progression.
巴雷特食管向癌症的进展是通过体内已被记录的细胞周期和基因异常的积累而发生的。为了更好地研究巴雷特食管的肿瘤演变，本研究的目的是从癌前组织建立体外培养体系。
采用机械和酶解方法从内镜活检标本中启动巴雷特上皮细胞培养，并通过流式细胞术、细胞遗传学和分子遗传学分析对细胞进行表征。
在39次尝试中建立了4个长期培养体系。所有培养体系均存在细胞遗传学异常以及流式细胞术检测到的4N DNA含量分数升高。检测到的分子遗传学异常包括：4个培养体系中的4个存在9p和/或CDKN2/p16异常，4个培养体系中的３个存在17p杂合性缺失和p53突变，4个培养体系中的1个存在5q杂合性缺失。p53失活与成功建立长期培养体系在统计学上相关。
这些培养体系包含细胞周期和分子遗传学异常，与先前在体内巴雷特食管癌症发生早期记录的异常情况非常相似。这些改变似乎也与体外成功生长有关。这些培养体系可能提供一个癌前体外系统，用于测试巴雷特食管的潜在治疗方法，以及研究肿瘤进展中重要的病因因素和基因中间体。
