人乳头瘤病毒阴性的喉鳞癌细胞系的建立  

Establishment of a HPV-negative cell line derived from xenografted human laryngeal squamous cell carcinoma

作　　者：蔡萍[1,2] 吴展元[1] 刘公汉[1] 李金荣[3] 华清泉[1] 詹汉章[1] 

机构地区：[1]武汉大学人民医院耳鼻咽喉头颈外科,430060 [2]武汉大学口腔医学院正畸科 [3]武汉大学口腔医学院口腔颌面外科

出　　处：《中华病理学杂志》2005年第8期533-536,共4页Chinese Journal of Pathology

摘　　要：目的建立1株人乳头状瘤病毒(HPV)阴性的喉鳞状细胞癌细胞系,为体外研究喉癌提供理想的实验模型。方法以逆转录聚合酶链反应(RTPCR)证实为HPV阴性的高分化喉鳞癌手术切除标本接种于裸鼠皮下,取连续传代2次的裸鼠皮下移植瘤进行体外原代培养。通过光镜、电镜、生长曲线、细胞周期时相、软琼脂克隆实验、异种移植成瘤实验、角蛋白、癌胚抗原及HPV检测,对其生物学特性进行初步分析。结果经裸鼠过渡所建立的高分化喉鳞癌细胞系(Lscc02)目前已传至86代,细胞生长增殖稳定。该细胞系呈单层形式生长,群体倍增时间为39.1h。透射电镜下见胞质内典型的张力原纤维,细胞间以桥粒方式连接。染色体为人类核型,呈亚三倍体,众数分布在63～72。该细胞系具有恶性肿瘤细胞生长特征:软琼脂中形成克隆,裸鼠接种成瘤且形态结构、分化程度与原发瘤相似。免疫组织化学显示高分子量细胞角蛋白及癌胚抗原阳性,PCR显示HPV阴性。结论建立Lscc02细胞系为研究无HPV感染喉癌的发生、发展规律及HPV与喉癌演进的关系提供了有价值的体外模型。Objective To establish a human laryngeal carcinoma cell line unassociated with human papillomavirus （HPV）. Methods Viable tissue of a well-differentiated laryngeal squamous cell carcinoma was obtained and tested negative for the presence of HPV by polymerase chain reaction. Minced tissue fragments were then transplanted subcutaneously into nude mice. After two successive passages, the tumor tissue was seeded into culture flasks and incubated in a medium containing 10% fetal bovine serum, insulin and epidermal growth factor. Tumor cell phenotype and molecular features were determined by various methods. Results A stable cell line, designated as Lscc-02, was successfully established after 86 culture passages. The cells grew as a monolayer with epithelioid features. The cell doubling time was approximately 39. 1 hours. The human origin of the tumor cells was confirmed by karyotype analysis. The squamous epithelial phenotype was demonstrated by the immunopositivity of anti-cytokeratin antibodies and ultrastructural presence of tonofilaments and desmosomes. The malignant nature of the cells was documented by their clonal formation in soft-agar and tumorigenicity in nude mice. Lscc-02 cells expressed carcinoembryonic antigen （CEA） and were negative for HPV DNA. Conclusion This newly established Lscc-02 laryngeal squamous cell carcinoma cell line may be a useful model for investigating laryngeal carcinoma unrelated to HPV infection, and the role of HPV in the progression of human laryngeal carcinoma.

关 键 词：喉 鳞状细胞癌 人 细胞系 HPV阴性 鳞癌细胞系 喉鳞状细胞 人乳头状瘤病毒(HPV) 人乳头瘤病毒 阴性 

分 类 号：R739.65[医药卫生—肿瘤]