title: Collagen gel protects L929 cells from TNFα-induced death by activating NF-κB.
abstract: OBJECTIVE
  Type I collagen is one of the most abundant components of extracellular matrix. We previously illustrated that murine fibrosarcoma L929 cells grew well on type I collagen gel and escaped from TNFα-induced cell death. In this study, we investigated the mechanism underlying the protective effect of collagen gel.
  
  
  METHODS
  We used western blot, confocal microscopy, MTT assay and flow cytometry by introducing fluorescence staining to determine the expression levels of nuclear factor kappa B (NF-κB), inhibitory ratio and autophagy.
  
  
  RESULTS
  L929 cells on collagen gel showed higher expression of NF-κB in the nucleus. Inhibition of NF-κB with pyrrolidine dithiocarbamate hydrochloride (PDTC) or knockdown by NF-κB-siRNA canceled the protective effect of collagen gel on L929 cells from TNFα-induced death, suggesting for the role of NF-κB in the protection from cell death. We found a new aspect of the effect of PDTC on L929 cells cultured on collagen gel. PDTC alone without TNFα induced apoptosis in the L929 cells cultured on collagen gel but not the cells on plastic dish. The apoptosis induction of the L929 cells cultured on collagen gel with PDTC was repressed by inhibiting autophagy with chloroquine, an autophagy inhibitor, suggesting that autophagy contributes to the death induced by the treatment with PDTC. Possible underlying mechanism of this finding is discussed.
  
  
  CONCLUSIONS
  NF-κB played an important role in protecting the L929 cells cultured on collagen gel from TNFα-induced death.
authors.affiliation: a China-Japan Research Institute of Medical and Pharmaceutical Sciences , Shenyang Pharmaceutical University , Shenyang , China.
authors.forename: Hong-Ju
authors.initials: HJ
authors.lastname: Wang
authors.affiliation: a China-Japan Research Institute of Medical and Pharmaceutical Sciences , Shenyang Pharmaceutical University , Shenyang , China.
authors.forename: Meng-Qi
authors.initials: MQ
authors.lastname: Li
authors.affiliation: a China-Japan Research Institute of Medical and Pharmaceutical Sciences , Shenyang Pharmaceutical University , Shenyang , China.
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Liu
authors.affiliation: a China-Japan Research Institute of Medical and Pharmaceutical Sciences , Shenyang Pharmaceutical University , Shenyang , China.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Hayashi
authors.affiliation: b Nippi Research Institute of Biomatrix, Nippi, Incorporated , Toride , Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Fujisaki
authors.affiliation: b Nippi Research Institute of Biomatrix, Nippi, Incorporated , Toride , Japan.
authors.forename: Shunji
authors.initials: S
authors.lastname: Hattori
authors.affiliation: c Department of Medical Education & Primary Care , Kyoto Prefectural University of Medicine , Kyoto , Japan.
authors.forename: Shin-Ichi
authors.initials: SI
authors.lastname: Tashiro
authors.affiliation: d Department of Clinical and Pharmaceutical Sciences , Showa Pharmaceutical University , Tokyo , Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Onodera
authors.affiliation: a China-Japan Research Institute of Medical and Pharmaceutical Sciences , Shenyang Pharmaceutical University , Shenyang , China.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ikejima
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D024042:Collagen Type I / Q000737:chemistry / Q000494:pharmacology; D020100:Hydrogels / Q000737:chemistry / Q000494:pharmacology; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D011392:Proline / Q000031:analogs & derivatives / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D024042:Collagen Type I; D020100:Hydrogels; D016328:NF-kappa B; D013859:Thiocarbamates; D014409:Tumor Necrosis Factor-alpha; C066229:prolinedithiocarbamate; D011392:Proline
keywords: Autophagy; NF-κB; PDTC; TNFα; collagen gel