Establishment and characterization of a tumorigenic cell line from areca quid and tobacco smoke-associated buccal carcinoma
Establishment and characterization of a tumorigenic cell line from areca quid and tobacco smoke-associated buccal carcinoma
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DOI：
10.1016/j.oraloncology.2006.07.007
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发表时间：
2007-08-01
期刊：
影响因子：
4.8
通讯作者：
Kuo, Mark Yen-Ping
中科院分区：
文献类型：
--
作者：
Kok, Sang-Heng;Hong, Chi-Yuan;Kuo, Mark Yen-Ping
从一位嚼槟榔、吸烟的48岁男性口腔颊粘膜原发性鳞状细胞癌的手术切除标本中，建立了一个细胞系TW 2.6。TW 2.6细胞具有角质形成细胞的形态特征，在培养中增殖迅速，倍增时间为24 h。核型显示人类染色体高度超二倍体和复杂的重排。Western blotting显示p53的显著表达和p21（CIP 1）的中等表达。p27（KIP 1）和p16（INK 4a）的基线表达几乎检测不到。TW 2.6细胞中Bax和Fas表达较低，Bcl-2表达较强。p53基因突变分析表明，在密码子220的第二个碱基处发生A → G的转换，导致蛋白质中的氨基酸由酪氨酸替换为半胱氨酸。功能分析表明，TW2.6不能激活p53特异性的p53启动子。Lipofectamine（TM）2000和磷酸钙沉淀技术为TW 2.6细胞提供了良好的转染效率，并可用于未来的转染实验。建立了TW2.6的异种移植物-SCID小鼠肿瘤模型。组织学检查表明，移植的肿瘤保持鳞状细胞癌的形态特征。结论：本研究成功建立了烟草相关性颊癌细胞系TW 2.6，为研究烟草相关性颊癌和AQ提供了一个有价值的模型。(C)2006爱思唯尔有限公司保留所有权利。
A cell tine, TW2.6, has been established from the surgically resected specimen of an untreated primary squamous cell carcinoma of the buccal mucosa from a 48-year-old man who was an areca quid chewer and tobacco smoker. TW2.6 cells exhibited morphological features of keratinocytes and replicated rapidly in culture with a doubling time of 24 h. The karyotype showed human chromosomes with high hyperdiploidy and complex rearrangements. Western blotting showed pronounced expression of p53 and moderate expression of p21(CIP1). The base-line expressions of p27(KIP1) and p16(INK4a) were barely detectable. Low levels of Bax and Fas were found in TW2.6 cells but Bcl-2 expression was more readily observed. Mutational analysis of p53 gene revealed an A -> G transition at the second base of codon 220, resulting in amino acid substitution from tyrosine to cysteine in the protein. Functional analysis showed that TW2.6 was unable to activate the p53-specific PUMA promoter. Lipofectamine(TM) 2000 and calcium phosphate precipitation technique offer good transfection efficiencies for TW2.6 cells and may be used in future transfection experiments. A xenograft-SCID mouse tumor model was established for TW2.6. Histological examination demonstrated that the engrafted tumors maintained the morphological features of a squamous cell carcinoma. It is thought that the establishment of tumorigenic TW2.6 cell line provides a valuable model for AQ and tobacco smoke-associated buccal carcinoma. (C) 2006 Elsevier Ltd. All rights reserved.