Isolation of differentiated squamous and undifferentiated spindle carcinoma cell lines with differing metastatic potential from a 4-nitroquinoline N-Oxide-induced tongue carcinoma in a F344 rat.
One differentiated squamous cell carcinoma (SCC) cell line (RSC3-E2) and two undifferentiated tumor cell lines (RSC3-LM and RSC3-E2R) with different metastatic potential were established from a 4-nitroquinoline N-oxide (4NQO)-induced differentiated SCC in F344 rat tongue. The RSC3-E2 subline was isolated from a parental cell line (RSC3-P) by single cell cloning in vitro, whereas the RSC3-LM subline was isolated from a lung metastatic focus after subcutaneous (s.c.) injection of RSC3-P cells. The RSC3-E2R cell line was isolated from a lung metastatic focus following s.c. injection of RSC3-E2 cells after X-irradiation in vitro. The RSC3-E2 cell line is keratin-positive and grows as a keratinizing tumor in nude mice, whereas RSC3-LM and RSC3-E2R cells are keratin-negative, vimentin-positive and form undifferentiated tumors. When s.c. injected into nude mice, the RSC3-E2 cell line proved to be non-metastatic, while the RSC3-LM cell line was metastatic by both hematogenous and lymphogenous routes, and the RSC3-E2R cell line was metastatic only hematogenously. In vitro relative growth rates and in vitro invasion activity of these cell lines were in the order RSC3-LM > RSC3-E2R > RSC3-E2. Chromosome analysis revealed two peaks with modal chromosome numbers of 83 and 78 for RSC3-P cells and single peaks at 83, 78 and 56 for RSC3-LM, RSC3-E2 and RSC3-E2R cell lines, respectively. Common structural abnormalities on chromosome 11 were shared by all cell lines. Mutation analysis of the p53 gene using a yeast functional assay demonstrated RSC3-LM cell line to have a point mutation at codon 269, whereas RSC3-E2 and RSC3-E2R had double mutations at codons 106 and 170 on each allele. These results suggest that the two undifferentiated RSC3-LM and RSC3-E2R tumor cell lines with different metastatic potential were generated from differentiated SCC cells via different genetic pathways as a consequence of tumor progression in vivo and in vitro, respectively. These cell lines should provide a useful model for understanding mechanisms of hematogenous and lymphogenous metastasis, as well as tumor progression of oral SCCs.
Takeuchi
S
S
Laboratory of Pathology, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya 464-8681, Japan. hnakanis@aichi-cc.pref.aichi.jp
Nakanishi
H
H
Yoshida
K
K
Yamamoto
S
S
Tonoki
H
H
Tsukamoto
T
T
Fukushima
S
S
Moriuchi
T
T
Kurita
K
K
Tatematsu
M
M
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity*; D000818:Animals; D002273:Carcinogens / Q000633:toxicity; D002277:Carcinoma / Q000139:chemically induced / Q000235:genetics / Q000473:pathology* / Q000556:secondary*; D002294:Carcinoma, Squamous Cell / Q000139:chemically induced / Q000235:genetics / Q000473:pathology* / Q000556:secondary*; D002999:Clone Cells; D007621:Karyotyping; D008175:Lung Neoplasms / Q000139:chemically induced / Q000235:genetics / Q000473:pathology / Q000556:secondary*; D008207:Lymphatic Metastasis; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009362:Neoplasm Metastasis; D009381:Neoplasms, Radiation-Induced / Q000235:genetics / Q000473:pathology; D051381:Rats; D011916:Rats, Inbred F344; D014059:Tongue / Q000187:drug effects / Q000473:pathology; D014062:Tongue Neoplasms / Q000139:chemically induced / Q000235:genetics / Q000473:pathology*; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D014965:X-Rays
D002273:Carcinogens; D015112:4-Nitroquinoline-1-oxide