title: Establishment of a stroma-dependent human acute myelomonocytic leukemia cell line, NAMO-2, with FLT3 tandem duplication.
abstract: We have established a stroma-dependent myelomonocytic cell line, NAMO-2, with FLT3 internal tandem duplication (FLT3/ITD). Leukemia cells from a patient with acute myelomonocytic leukemia were administered to form subcutaneous tumors in nude mice, which were maintained successively, although we failed to establish continuously growing cells from the original leukemia cell culture. In the cultures of cells from subcutaneous tumors, there were stroma cells that had originated from the nude mice and showed continuous growth. The leukemia cells showed continuous growth dependent on this stroma, and this cell line was named NAMO-2. Detection of FLT3/ITD by the reverse transcriptase polymerase chain reaction (PCR) and genomic PCR showed that NAMO-2 was homozygous for FLT3/ITD. Constitutive activation of FLT3 was detected by Western blotting, and the phosphorylation of Akt, MEK, and STAT5 was also observed. FLT3 kinase inhibitor AG1296 specifically inhibited cell growth. NAMO-2 provides a useful tool to analyze adherence-dependent survival signaling of leukemia with FLT3/ITD and a model for the screening of FLT3 kinase inhibitors.
authors.affiliation: Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya, Japan. aakihiro@med.nagoya-u.ac.jp
authors.forename: Akihiro
authors.initials: A
authors.lastname: Abe
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kiyoi
authors.forename: Manabu
authors.initials: M
authors.lastname: Ninomiya
authors.forename: Tomio
authors.initials: T
authors.lastname: Yamazaki
authors.forename: Takuhei
authors.initials: T
authors.lastname: Murase
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Ozeki
authors.forename: Momoko
authors.initials: M
authors.lastname: Suzuki
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Hayakawa
authors.forename: Akira
authors.initials: A
authors.lastname: Katsumi
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Emi
authors.forename: Tomoki
authors.initials: T
authors.lastname: Naoe
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D006720:Homozygote; D006801:Humans; D015479:Leukemia, Myelomonocytic, Acute / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental; D010766:Phosphorylation; D015398:Signal Transduction; D017154:Stromal Cells / Q000502:physiology*; D020080:Tandem Repeat Sequences; D014183:Transplantation, Heterologous; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics*
chemical_list: C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3