Witjes M, Scholma J, van Drunen E, Roodenburg J L, Mesander G, Hagemeijer A, Tomson A M
Department of Oral and Maxillofacial Surgery, University Hospital Groningen, The Netherlands.
Carcinogenesis. 1995 Nov;16(11):2825-32. doi: 10.1093/carcin/16.11.2825.
This study describes several characteristics of a cell line, UHG-RaC '93 derived from rat oral squamous cell carcinoma induced by the carcinogen 4-nitroquinoline-1-oxide (4NQO). The cell line was established from explant cultures without support of fibroblast feeder cells and continued for > 30 passages. UHG-RaC '93 had a high mitotic rate with a population doubling time of 25 h and a high rate of squame production. The first passage had a low colony-forming efficiency in agarose gel, whereas later passages did not grow at all in semi-solid medium. Phenotype selection was furthermore apparent from a gradual increase of the trypsin-detachment time. Cytogenetic analysis showed that UHG-RaC '93 was hypotetraploid with an average of 74 chromosomes. Abnormalities compared to the normal karyotype were assessed and consisted mainly of breakpoints at (1)(q5?3), (3)(p1), (3)(q11q23), (11)(p?11), (13)(p13) and a derivative (12)t(12;13)(q10;q10). The karyotype remained stable for at least 26 passages. The expression of typical epithelioid markers like cytokeratins and desmoglein corresponded with normal rat oral keratinocytes. However expression of alpha 6 beta 4-integrin was altered. Squame production, immunophenotype and anchorage dependency indicated that UHG-RaC '93 had the same features of a well-differentiated carcinoma with a low degree of agressiveness as the original tumour. The stable karyotype of this cell line provides a basis for further analysis of the effect of 4NQO on the genotype, phenotype and behaviour of rat oral keratinocytes.
本研究描述了一种细胞系UHG-RaC '93的几个特征,该细胞系源自由致癌物4-硝基喹啉-1-氧化物(4NQO)诱导的大鼠口腔鳞状细胞癌。该细胞系是在没有成纤维细胞饲养层细胞支持的情况下从外植体培养物中建立的,并持续传代超过30次。UHG-RaC '93具有高有丝分裂率,群体倍增时间为25小时,且鳞状细胞产生率高。第一代在琼脂糖凝胶中的集落形成效率较低,而后续传代在半固体培养基中根本不生长。此外,从胰蛋白酶消化时间的逐渐增加可以明显看出表型选择。细胞遗传学分析表明,UHG-RaC '93是亚四倍体,平均有74条染色体。评估了与正常核型相比的异常情况,主要包括(1)(q5?3)、(3)(p1)、(3)(q11q23)、(11)(p?11)、(13)(p13)处的断点以及一个衍生的(12)t(12;13)(q10;q10)。核型至少在26次传代中保持稳定。细胞角蛋白和桥粒芯糖蛋白等典型上皮样标志物的表达与正常大鼠口腔角质形成细胞一致。然而,α6β4整合素的表达发生了改变。鳞状细胞产生、免疫表型和锚定依赖性表明,UHG-RaC '93具有与原始肿瘤相同的低侵袭性高分化癌的特征。该细胞系稳定的核型为进一步分析4NQO对大鼠口腔角质形成细胞的基因型、表型和行为的影响提供了基础。