Article

Ganglioside GD1a Negatively Regulates Matrix Metalloproteinase-9 Expression in Mouse FBJ Cell Lines at the Transcriptional Level

Laboratory of Tumor Biology and Glycobiology, Shenyang Pharmaceutical University, Shenyang, P. R. China.
Connective Tissue Research (Impact Factor: 1.98). 02/2007; 48(4):198-205. DOI: 10.1080/03008200701458731
Source: PubMed

ABSTRACT Mouse FBJ virus-induced osteosarcoma FBJ-S1 cells rich in GD1a are not readily metastatic, whereas FBJ-LL cells with low levels of GD1a are highly metastatic. GD1a was previously shown to suppress metastasis of mouse FBJ cells and to upregulate caveolin-1 and stromal interaction molecule 1 expression. The present study demonstrates that matrix metalloproteinase-9 (MMP-9) expression renders FBJ-LL cells invasive. MMP-9 is inversely regulated by GD1a, based upon four observations: MMP-9 mRNA content was 5 times higher in FBJ-LL cells than FBJ-S1 cells; a GD1a-re-expressing FBJ-LL cell variant produced through beta1,4GalNAcT-1 cDNA transfection expressed lower levels of MMP-9; exogenous addition of GD1a to FBJ-LL cells decreased MMP-9 production in a dose- and time-dependent manner; and treatment of GD1a-rich cells with D-PDMP or siRNA targeting St3gal2 decreased GD1a expression, but augmented MMP-9 expression. This is the first report demonstrating that GD1a negatively regulates expression of MMP-9 at the transcriptional level.

Download full-text

Full-text

Available from: Tatsuya Yamagata, Aug 28, 2015
0 Followers
 · 
126 Views
  • Source
    • "The poorly metastatic osteosarcoma cell line FBJ-S1, which is produced from FBJ virusinduced osteosarcoma in the BALB/c mouse, expresses gangliosides GM3 and GD1a, whereas the highly metastatic FBJ-LL cell line expresses GM3 but is virtually deficient in GD1a [18]. GD1a has been shown to inversely regulate the metastatic capability of FBJ cells [19] through enhanced expression of Cav-1 and stromal interaction molecule 1 (Stim1) [20] and the suppression of MMP-9 [21], TNF-α [22], and NOS2 [23]. HGF binds to and phosphorylates MET for its activation [7]. "
    Dataset: 2011BBA HGF
  • Source
    • "Cells were cultured as reported previously [8]. Cell proliferation was measured using the WST-1 assay [9] [11] with a cell counting kit (Dojindo, Kumamoto, Japan) according to the manufacturer's protocol. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Caveolin-1, an indispensable component of caveolae serving as a transformation suppressor protein, is highly expressed in poorly metastatic mouse osteosarcoma FBJ-S1 cells while highly metastatic FBJ-LL cells express low levels of caveolin-1. Calcium concentration is higher in FBJ-S1 cells than in FBJ-LL cells; therefore, we investigated the possibility that calcium signaling positively regulates caveolin-1 in mouse FBJ-S1 cells. When cells were treated with the calcium channel blocker nifedipine, cyclosporin A (a calcineurin inhibitor), or INCA-6 (a nuclear factor of activated T-cells [NFAT] inhibitor), caveolin-1 expression at the mRNA and protein levels decreased. RNA silencing of voltage-dependent L-type calcium channel subunit alpha-1C resulted in suppression of caveolin-1 expression. This novel caveolin-1 regulation pathway was also identified in mouse NIH 3T3 cells and Lewis lung carcinoma cells. These results indicate that caveolin-1 is positively regulated at the transcriptional level through a novel calcium signaling pathway mediated by L-type calcium channel/Ca(2+)/calcineurin/NFAT.
    Biochemical and Biophysical Research Communications 08/2012; 426(3):334-41. DOI:10.1016/j.bbrc.2012.08.079 · 2.28 Impact Factor
  • Source
    • "The poorly metastatic osteosarcoma cell line FBJ-S1, which is produced from FBJ virusinduced osteosarcoma in the BALB/c mouse, expresses gangliosides GM3 and GD1a, whereas the highly metastatic FBJ-LL cell line expresses GM3 but is virtually deficient in GD1a [18]. GD1a has been shown to inversely regulate the metastatic capability of FBJ cells [19] through enhanced expression of Cav-1 and stromal interaction molecule 1 (Stim1) [20] and the suppression of MMP-9 [21], TNF-α [22], and NOS2 [23]. HGF binds to and phosphorylates MET for its activation [7]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: BACKGROUND: Hepatocyte growth factor (HGF) is a mesenchyme-derived, multifunctional protein that is implicated in tumor growth and invasive behavior. Some tumor cells express both HGF and its receptor MET, forming an autocrine loop that permanently activates it. Ganglioside GD1a suppresses metastatic capacity in murine FBJ osteosarcoma cells and MET phosphorylation activated by HGF binding, but the signaling pathway controlling HGF production has not been fully explored. METHODS: Expression of HGF, caveolins, or MET of the cells that had been transfected with siRNA or cDNA directed to GM2/GD2 synthase, caveolin-1 or HGF was determined by semi-quantitative RT-PCR and Western blots. RESULTS: HGF expression in highly metastatic, GD1a-deficient FBJ-LL cells was higher than that in the poorly metastatic, GD1a-rich FBJ-S1 cells. Transfection with GM2/GD2 synthase cDNA increased GD1a levels in FBJ-LL cells and suppressed HGF expression. Treatment with siRNAs directed toward GM2/GD2 synthase in FBJ-S1 cells reduced gangliosides and augmented HGF expression. GD1a was found to be the only ganglioside species suppressing HGF expression upon addition to FBJ-LL cells. HGF expression was decreased by GD1a addition to FBJ-LL cells after 48h, enough to induce caveolin-1 expression. Silencing caveolin-1 up-regulated HGF, and the re-introduction of caveolin-1 cDNA decreased HGF expression. Caveolin-1 suppressed MET phosphorylation. We also found GD1a regulation of HGF in Lewis lung carcinoma cells. CONCLUSIONS: HGF expression was negatively regulated by GD1a through caveolin-1 at the transcriptional level via the suppression of MET phosphorylation. GENERAL SIGNIFICANCE: This is the first report that ganglioside GD1a negatively regulates HGF expression through caveolin-1.
Show more