Characterization of newly established tumor lines from a spontaneous malignant schwannoma in F344 rats: nerve growth factor production, growth inhibition by transforming growth factor-beta1, and macrophage-like phenotype expression.
Transplantable tumor (KE) and clone cell (KE-F11) lines were established from a spontaneous malignant schwannoma found in an aged F344 rat. The primary tumor and KE tumors consisted of oval or spindle cells arranged in ill-defined bundles. Cultured KE-F11 cells exhibited polygonal or spindle configurations. Immunohistochemically, neoplastic cells in KE and KE-F11 reacted to vimentin, S-100 protein, neuron-specific enolase, myelin basic protein, and glial fibrillary acidic protein in varying degrees, indicating neurogenic features; occasional cells reacted to alpha-smooth muscle actin. Cells positive for lysosomal enzymes (acid phosphatase and non-specific esterase), and ED1 (rat macrophage specific) were observed in KE-F11, and electron microscopically, cells with many lysosomes were frequently present, indicating expression of macrophage-like phenotypes. Bioassay analysis revealed that KE-F11 cells produced high levels of nerve growth factor. DNA synthesis was inhibited by addition of transforming growth factor-beta1 (TGF-beta1), and Northern blot analysis revealed that expression of c-myc, a cell cycle-related immediate early gene, was depressed by TGF-beta1. Likely, TGF-beta1 is a factor capable of inhibiting cellular growth of Schwann cells. mRNA expression of the low-density lipoprotein receptor-related protein (LRP) was seen in KE-F11 cells by Northern blot analysis, and the level was decreased by lipopolysaccharide (LPS) treatment. LRP may be attributable to regulation of Schwann cell functions. KE-F11 cells seeded on laminin-coated dishes exhibited more extended cytoplasmic projections than on collagen type I-coated dishes. The present study provides evidence that biological properties of malignant schwannoma-derived cells might be affected by exogenous factors such as TGF-beta1, LPS and laminin. These tumor lines may be useful for studies on pathobiological characteristics of Schwann cells.
Yamate
J
J
Laboratories of Veterinary Pathology and Anatomy, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, Gakuencho 1-1, Sakai, 599-8531 Osaka, Japan. yamate@vet.osakafu-u.ac.jp
Yasui
H
H
Benn
S J
SJ
Tsukamoto
Y
Y
Kuwamura
M
M
Kumagai
D
D
Sakuma
S
S
LaMarre
J
J
D000199:Actins / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D002452:Cell Count; D002461:Cell Line, Transformed; D003893:Desmin / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D016758:Genes, jun / Q000502:physiology; D016259:Genes, myc / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D066298:In Vitro Techniques; D007633:Keratins / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000473:pathology / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D004676:Myelin Basic Protein / Q000378:metabolism; D020932:Nerve Growth Factor / Q000378:metabolism*; D009442:Neurilemmoma / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D016716:PC12 Cells; D010641:Phenotype; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011916:Rats, Inbred F344; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D009418:S100 Proteins / Q000378:metabolism; D013194:Staining and Labeling; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D053773:Transforming Growth Factor beta1; D014746:Vimentin / Q000378:metabolism
D000199:Actins; D003893:Desmin; D005904:Glial Fibrillary Acidic Protein; D008070:Lipopolysaccharides; D004676:Myelin Basic Protein; D012333:RNA, Messenger; D009418:S100 Proteins; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D014746:Vimentin; D007633:Keratins; D020932:Nerve Growth Factor; D010751:Phosphopyruvate Hydratase