title: [Establishment of a HPV-negative cell line derived from xenografted human laryngeal squamous cell carcinoma].
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Otolaryngology- Head and Neck Surgery, Renmin Hospital, Wuhan University, Wuhan 430060, China. caip@eyou.com
authors.forename: Ping
authors.initials: P
authors.lastname: Cai
authors.forename: Zhan-Yuan
authors.initials: ZY
authors.lastname: Wu
authors.forename: Jin-Rong
authors.initials: JR
authors.lastname: Li
authors.forename: Gong-Han
authors.initials: GH
authors.lastname: Liu
authors.forename: Qing-Quan
authors.initials: QQ
authors.lastname: Hua
authors.forename: Han-Zhang
authors.initials: HZ
authors.lastname: Zhan
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000473:pathology* / Q000821:virology; D045744:Cell Line, Tumor; D006801:Humans; D007822:Laryngeal Neoplasms / Q000473:pathology* / Q000821:virology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D027383:Papillomaviridae