title: Characterization of cells cultured from chylous effusion from a patient with sporadic lymphangioleiomyomatosis.
abstract: BACKGROUND
  Lymphangioleiomyomatosis (LAM) is a progressive, rare interstitial lung disease that almost exclusively affects women. It is caused by a mutation in one of the tuberous sclerosis genes, TSC1 or TSC2, and constitutive activation of the mammalian target of rapamycin (mTOR) pathway in smooth muscle-like cells (LAM cells). The heightened proliferation and accumulation of LAM cells leads to the destruction of lung tissue.
  
  
  METHODS
  In the present study, we developed a cell line (S-LAM1) derived from a chylous effusion obtained from a patient with sporadic, pulmonary LAM and evaluated its phenotype using immunofluorescence, flow cytometry, and an image stream system. Ultrastructure was assessed using a transmission electron microscope. To assess the ability of LAM cells to move and migrate (which is strictly associated with the ability to metastasize), we carried-out a real-time polymerase chain reaction (PCR) array analysis of 84 genes involved in cell motility. In order to evaluate the effect of rapamycin, a natural inhibitor of mTOR kinase, on S-LAM1 cells, a sulforhodamine B cell viability assay was performed with different concentrations of rapamycin.
  
  
  CONCLUSIONS
  The phenotype of these cells is consistent with the biology of LAM cells. S-LAM1 cells present combined smooth muscle, melanocytic, and lymphatic endothelium lineage, as well as the presence of mesenchymal differentiation markers. A particular pattern of gene expression, including high expression of ezrin (EZR), myosin heavy chain 10, non-muscle (MYH10), and myosin light chain kinase (MYLK) and a greatly decreased expression of supervillin (SVIL), when compared to controls, indicates a high potential motility activity, especially of cell spreading. Rapamycin significantly, although only partially, inhibited S-LAM1 cell proliferation in vitro, and should, perhaps, be considered in the future in combination with other agents.
authors.affiliation: Department of Histology and Embryology, Wroclaw Medical University, Wroclaw, Poland irminagrzegorek@o2.pl.
authors.forename: Irmina
authors.initials: I
authors.lastname: Grzegorek
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Zuba-Surma
authors.affiliation: Department of Surgery, Fourth Military Hospital, Wroclaw, Poland Department of Clinical Proceedings, Wroclaw Medical University, Wroclaw, Poland.
authors.forename: Mariusz
authors.initials: M
authors.lastname: Chabowski
authors.affiliation: Department of Surgery, Fourth Military Hospital, Wroclaw, Poland Department of Clinical Proceedings, Wroclaw Medical University, Wroclaw, Poland.
authors.forename: Dariusz
authors.initials: D
authors.lastname: Janczak
authors.affiliation: Department of Internal Medicine, Fourth Military Hospital, Wroclaw, Poland Department of Angiology, Faculty of Health Science, Wroclaw Medical University, Wroclaw, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Szuba
authors.affiliation: Department of Histology and Embryology, Wroclaw Medical University, Wroclaw, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Dziegiel
mesh_terms: D000328:Adult; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D018192:Lymphangioleiomyomatosis / Q000235:genetics* / Q000473:pathology; D051379:Mice; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009362:Neoplasm Metastasis; D020123:Sirolimus / Q000008:administration & dosage*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: C000624650:TSC1 protein, human; C000624653:TSC2 protein, human; C000624651:Tsc1 protein, mouse; C000624654:Tsc2 protein, mouse; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus
keywords: LAM cells; Lymphangioleiomyomatosis; cell motility; mTOR; metastasis; rapamycin