Multiple genotypic aberrances associate to terminal differentiation-deficiency of an oral squamous cell carcinoma in serum-free culture

• Published in: Differentiation (London) 2008-10, Vol.76 (8), p.868-880
• Main Authors: Roberg, Karin, Ceder, Rebecca, Farnebo, Lovisa, Norberg-Spaak, Lena, Grafström, Roland C.
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis - genetics; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - pathology; Cell Differentiation - genetics; Cell Line, Transformed; Cell Line, Tumor; Cell Proliferation; Culture Media, Serum-Free; Female; Genotype; growth and terminal differentiation; Humans; Keratinocytes - cytology; MEDICIN; Medicin och hälsovetenskap; MEDICINE; Mice; Mice, Inbred BALB C; Mice, Nude; Middle Aged; Mouth Neoplasms - genetics; Mouth Neoplasms - pathology; normal epithelial cells; oral squamous cell carcinoma; serum-free culture; transcript profiling for tumor markers
• ISSN: 0301-4681; 1432-0436; 1432-0436
• DOI: 10.1111/j.1432-0436.2008.00267.x

Oral squamous cell carcinoma (OSCC) lines proliferative in the serum-free conditions devised for normal oral keratinocytes (NOK) are virtually absent, complicating studies of carcinogenesis. A tongue squamous cell carcinoma generated under conditions for normal cell culture an apparently immortal line (termed LK0412) that has undergone ≥200 population doublings from over a year in culture. LK0412 exhibited epithelial morphology, intermediate filaments, desmosomes, and cytokeratin. Soft agar growth and tumorigenicity in athymic nude mice indicated the malignant phenotype. Compared with NOK, LK0412 exhibited increased indices for proliferation and apoptosis, and a decreased terminal differentiation index. Fetal bovine serum inhibited growth and increased apoptosis but failed to induce terminal differentiation of LK0412; the latter outcome differed clearly from that in NOK. Gene ontology assessment of transcript profiles implicated multiple alterations in biological processes, molecular functions, and cellular components in LK0412. Genetic changes, some that were confirmed to the protein level, included previously proposed OSCC markers, i.e., BAX, CDC2, and TP53, as well as multiple cancer-associated genes not considered for OSCC, e.g., BST2, CRIP1, ISG15, KLRC1, NEDD9, NNMT, and TWIST1. Elevation of p53 protein agreed with a missense mutation detectable in both the LK0412 line and the original tumor specimen. Moderate differentiation characterized the original tumor as well as tumors generated from inoculation of LK0412 in mice. Overall, the results suggest that the LK0412 cell line represent a subgroup of OSCC with unique genomic and phenotypic profiles. LK0412 should be useful to exploration of OSCC development, particularly the deregulated growth and differentiation responsiveness to serum factors.