Tannehill-Gregg S, Kergosien E, Rosol T J
Department of Veterinary Biosciences, The Ohio State University, Columbus 43210, USA.
In Vitro Cell Dev Biol Anim. 2001 Nov-Dec;37(10):676-83. doi: 10.1290/1071-2690(2001)037<0676:FHANSC>2.0.CO;2.
A natural animal model for human head and neck squamous cell carcinoma (H/N SCC) has not been described. The domestic cat has a high spontaneous occurrence of oropharyngeal SCC, which is similar to the human disease in aggressiveness and incurability. We have developed a cell line (SCCF1) from a laryngeal SCC of a cat. Keratinocytes were maintained in culture for greater than 50 passages. SCCF1 had strong cytokeratin immunohistochemical staining, weak vimentin staining, and no p53 staining. Ultrastructual features included cytokeratin filaments and desmosomes, as well as features of anaplasia (irregular cytoplasmic and nuclear margins, surface filopodia, and abnormal intermediate filament production). Karyotype analysis revealed aneuploidy, with a stemline chromosomal number of 34. The cells grew logarithmically for 6 d until confluency. SCCF1 expressed parathyroid hormone-related protein (PTHrP) messenger ribonucleic acid (mRNA) and protein, and secreted the protein into the medium. Treatment of SCCF1 with transforming growth factor-beta increased PTHrP production but did not affect PTHrP mRNA stability. Reverse transcriptase-polymerase chain reaction was used to amplify a 282-base pair region of feline PTHrP mRNA, encoding portions of the pre-pro and coding regions. The complementary deoxyribonucleic acid (cDNA) was cloned and sequenced. The cDNA and the predicted amino acid sequences had a high degree of homology to human and canine PTHrP. RT-PCR was used to confirm alternate splicing of PTHrP mRNA for translation of PTHrP 1-139 and PTHrP 1-141. The SCCF1 cell line will permit mechanistic experiments on genetic dysregulation in neoplastic keratinocytes of the feline oropharynx, and development of an in vitro model for H/N cancer.
尚未有用于人类头颈部鳞状细胞癌(H/N SCC)的天然动物模型被描述。家猫口咽鳞状细胞癌的自发发生率很高,其侵袭性和难治性与人类疾病相似。我们从一只猫的喉鳞状细胞癌中建立了一个细胞系(SCCF1)。角质形成细胞在培养中维持了50代以上。SCCF1细胞角蛋白免疫组化染色强,波形蛋白染色弱,p53染色阴性。超微结构特征包括角蛋白丝和桥粒,以及间变特征(不规则的细胞质和核边缘、表面丝状伪足和异常的中间丝产生)。核型分析显示为非整倍体,干细胞系染色体数为34。细胞对数生长6天直至汇合。SCCF1表达甲状旁腺激素相关蛋白(PTHrP)信使核糖核酸(mRNA)和蛋白,并将该蛋白分泌到培养基中。用转化生长因子-β处理SCCF1可增加PTHrP的产生,但不影响PTHrP mRNA的稳定性。逆转录聚合酶链反应用于扩增猫PTHrP mRNA的一个282碱基对区域,该区域编码前原区和编码区的部分。互补脱氧核糖核酸(cDNA)被克隆并测序。该cDNA和预测的氨基酸序列与人和犬的PTHrP具有高度同源性。逆转录-聚合酶链反应(RT-PCR)用于确认PTHrP mRNA的可变剪接,以翻译PTHrP 1-139和PTHrP 1-141。SCCF1细胞系将允许对猫口咽肿瘤性角质形成细胞的基因失调进行机制实验,并开发一种H/N癌症的体外模型。