An experimental model of multistep laryngeal carcinogenesis: combined effect of human papillomavirus type 16 genome and N-methyl-N'-nitro-N-nitrosoguanidine.
This report describes the two-step in vitro transformation of human laryngeal epithelial cells (HLEC cells). Primary cultured HLEC cells were first transfected with cloned full-length human papillomavirus type 16 DNA, and two immortalized cell lines (HLEC-16 cell lines) were selected by subculturing transfected cells that continued to proliferate. The HLEC-16 cell lines were not tumorigenic in nude mice, and did not proliferate well in a culture medium containing a physiological level of calcium (Dullbecco's minimum essential medium supplemented with 10% fetal bovine serum = DMEM + 10%FBS). The HLEC-16 cell lines were secondarily exposed to N-methyl-N'-nitro-N-nitrosoguanidine, and several proliferating colonies were isolated in DMEM + 10%FBS. Among these calcium/serum-resistant cell colonies, one colony exhibited enhanced proliferation capacity in nude mice. These results support the hypothesis that human laryngeal epithelial cells may be the target for neoplastic transformation by a combined effect of human papillomaviruses and chemical carcinogens.
Tsutsumi
K
K
Department of Ontolaryngology, St. Marianna University School of Medicine, Kawasaki, Japan.
Iwatake
H
H
Suzuki
T
T
D000818:Animals; D014408:Biomarkers, Tumor; D018929:Cell Culture Techniques; D002465:Cell Movement; D004279:DNA, Viral; D016158:Genes, p53; D006801:Humans; D007830:Larynx / Q000187:drug effects* / Q000648:ultrastructure* / Q000821:virology; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology*; D051379:Mice; D027383:Papillomaviridae / Q000302:isolation & purification*; D012367:RNA, Viral
D014408:Biomarkers, Tumor; D004279:DNA, Viral; D012367:RNA, Viral; D008769:Methylnitronitrosoguanidine