Bikunin Target Genes in Ovarian Cancer Cells Identified by Microarray Analysis

Suzuki, Mika; Kobayashi, Hiroshi; Tanaka, Yoshiko; Hirashima, Yasuyuki; Kanayama, Naohiro; Takei, Yuji; Saga, Yasushi; Suzuki, Mitsuaki; Itoh, Hiroshi; Terao, Toshihiko

doi:10.1074/jbc.m300239200

Cited by 47 publications

(61 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The link between PI3K and protein kinase C has been recently elucidated (43). Our recent data showed that, using cDNA microarray analysis, PI3K may be a candidate of target genes for bikunin, a Kunitz-type protease inhibitor, also known as an antimetastatic compound (44), and that bikunin significantly inhibits translocation of protein kinase C from plasma membrane to cytoplasm, suggesting a cross-communication between PI3K and protein kinase C (23).…”

Section: Discussionmentioning

confidence: 99%

Transforming Growth Factor-β1-dependent Urokinase Up-regulation and Promotion of Invasion Are Involved in Src-MAPK-dependent Signaling in Human Ovarian Cancer Cells

Tanaka

Kobayashi

Suzuki

et al. 2004

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Urokinase-type plasminogen activator (uPA) has been implicated in tumor cell invasion and metastasis. We reported previously that transforming growth factor (TGF)-␤1 induces a dose-and time-dependent up-regulation of uPA mRNA and protein in highly invasive human ovarian cancer cell line HRA, leading to invasion. To further elucidate the mechanism of the invasive effect of TGF-␤1, we investigated which signaling pathway transduced by TGF-␤1 is responsible for this effect. Here, we show that 1) nontoxic concentrations of TGF-␤1 activated Src kinase; 2) TGF-␤1 rapidly phosphorylates ERK1/2 and Akt, but not p38; 3) pharmacological Src inhibitor PP2 or antisense (AS) c-Src oligodeoxynucleotide (ODN) treatment reduced TGF-␤1-induced phosphorylation of ERK1/2 and Akt by 85-90% compared with controls; 4) pharmacological inhibition of MAPK by PD98059 abrogated TGF-␤1-mediated Akt stimulation, whereas TGF-␤1-induced ERK1/2 stimulation was not inhibited by PI3K inhibitor LY294002 or AS-PI3K ODN transfection; 5) up-regulation of uPA mRNA in response to TGF-␤1 was almost totally blocked by PP2 and PD98059 and partially (ϳ55%) by LY294002; 6) TGF-␤1-induced uPA mRNA up-regulation was inhibited by treatment with AS ODNs to c-Src or PI3K by 90 or 60%, respectively, compared with control ODN treatment; and 7) blockade of the release of the transcription factor NF-B by pyrrolidinedithiocarbamate reduced the TGF-␤1-induced activation of the uPA gene by ϳ65%. In addition, curcumin, a blocker of the transcriptional factor AP-1, partially (35%) canceled this effect. Taken together, these data support a role for TGF-␤1 activation of two distinct pathways (Src-MAPK-PI3K-NF-B-dependent and Src-MAPK-AP-1-dependent) for TGF-␤1-dependent uPA up-regulation and promotion of invasion.The processes of ovarian cancer dissemination are characterized by altered local proteolysis, cellular proliferation, cell attachment, and invasion, suggesting that the urokinase-type plasminogen activator (uPA) 1 could be involved in the pathogenesis of peritoneal dissemination (1). uPA is a serine protease associated with various pathological conditions including tumor invasion and metastasis (2). One of the factors regulating the metastatic process is considered to be transforming growth factor-␤ (TGF-␤), which is a multifunctional cytokine that elicits numerous cellular effects pertinent to the metastatic process (1). TGF-␤ regulates a wide range of physiological and pathological cellular processes, including cell growth, differentiation, invasion, migration, mesenchymal transition, extracellular matrix synthesis, and cell death in many cell types including ovarian cancer cells (3). Recent data demonstrated that TGF-␤ specifically stimulates up-regulation of uPA mRNA and protein in certain types of neoplastic cells (1). The cellular mechanism(s) of the TGF-␤-induced uPA-dependent tumor invasion and metastasis has been extensively studied. The identity of the signaling pathway(s) involved in the TGF-␤-induced uPA up-regulation (4) remains less known. In a well...

show abstract

Section: Discussionmentioning

confidence: 99%

Transforming Growth Factor-β1-dependent Urokinase Up-regulation and Promotion of Invasion Are Involved in Src-MAPK-dependent Signaling in Human Ovarian Cancer Cells

Tanaka

Kobayashi

Suzuki

et al. 2004

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Initially the cDNA microarrays (TaKaRa Mouse IntelliGene II; TaKaRa Bio Inc., Otsu, Japan; http://www.takara-bio.co.jp) were spotted with 4277 sequence-verified clones that are available from GenBank (http://www.ncbi.nlm.nih.gov/Genbank/index.html). Probes for cDNA microarrays were generated using total RNA from ovaries in a standard reverse transcriptase reaction in which some of the dTTP was replaced with either Cy3-labeled dUTP or Cy5-labeled dUTP (Suzuki et al 2003b). In some experiments, the sample was labeled with Cy3; in others, it was labeled with Cy5, with essentially identical results.…”

Section: Microarray Probe Labeling and Hybridizationmentioning

confidence: 99%

“…To determine the fluorescent intensities of the two dyes for each spot, the fluorescence signals of Cy3-and Cy5-tagged cDNA spots on arrays at 532 nm (Cy3) and 635 nm (Cy5) were scanned simultaneously and immediately with an Affymetrix 428 Array Scanner and quantitated using a BioDiscovery ImaGene v. 4·2 (TaKaRa Bio Inc.) (Suzuki et al 2003b). The background-subtracted median ratio value was calculated for each spot, and replicate spots on each slide were averaged.…”

Section: Microarray Data Analysismentioning

confidence: 99%

Reproductive failure in mice lacking inter-alpha-trypsin inhibitor (ITI) – ITI target genes in mouse ovary identified by microarray analysis

Suzuki¹,

Kobayashi²,

Tanaka³

et al. 2004

Journal of Endocrinology

Self Cite

View full text Add to dashboard Cite

Bikunin, a Kunitz-type protease inhibitor, is found in blood and urine. It has been established by two laboratories independently that the bikunin knockout female mice display a severe reduction in fertility: the cumulus oophorus has a defect in forming the extracellular hyaluronanrich matrix during expansion. Proteins of the inter-alphatrypsin inhibitor (ITI) family are eliminated in mice in which the bikunin gene has been inactivated, since bikunin is essential for their biosynthesis. Proteins of the ITI family may contribute to the microenvironment in which ovulation takes place. It is not clear, however, whether a single mechanism affects the reproductive function including ovulation. For identifying the full repertoire of the ITI deficiency-related genes, a cDNA microarray hybridization screening was conducted using mRNA from ovaries of wild-type or bik / female mice. A number of genes were identified and their regulation was confirmed by real-time RT-PCR analysis. Our screen identified that 29 (0·7%) and 5 genes (0·1%) of the genes assayed were, respectively, up-and down-regulated twofold or more. The identified genes can be classified into distinct subsets. These include stress-related, apoptosis-related, proteases, signaling molecules, agingrelated, cytokines, hyaluronan metabolism and signaling, reactive oxygen species-related, and retinoid metabolism, which have previously been implicated in enhancing follicle development and/or ovulation. Real-time RT-PCR analysis confirmed that these genes were up-and down-regulated two-to tenfold by bikunin knockout. These studies demonstrate that proteins of the ITI family may exert potent regulatory effects on a major physiological reproductive process, ovulation.

show abstract

“…1. Bikunin suppresses expression of pro-inflammatory cytokines induced by LPS in mouse neutrophils (lanes 1-7) and human neutrophils (lanes [8][9][10]. Neutrophils were incubated with control medium or with the indicated concentrations of bikunin for 1 h, then cultured with or without LPS (1 µg/ml) for 90 min.…”

Section: Western Blot Analysismentioning

confidence: 99%

“…8 The broad spectrum of the functions of bikunin suggests the existence of multiple molecular targets that mediate diverse responses to the compounds in cells. 9 Current understanding of the mechanism of action by bikunin comes mostly from studies on the suppression of several signaling cascades. 10 Upon exposure to bikunin, abrogation of calcium (Ca 2+ ) influx modulates the transcription of target genes through mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K) dependent transcription.…”

Section: Introductionmentioning

confidence: 99%

Bikunin suppresses expression of pro-inflammatory cytokines induced by lipopolysaccharide in neutrophils

Kanayama

Yamada

Onogi

et al. 2007

Journal of Endotoxin Research

Self Cite

View full text Add to dashboard Cite

Activated neutrophils contribute to the development of preterm delivery. Because of its ability to suppress inflammation, bikunin, a Kunitz-type protease inhibitor, is currently in clinical trials. To investigate the molecular mechanism of this inhibition, we analyzed the effect of bikunin on proinflammatory cytokine production and nuclear factor-kappaB (NF-κB) activation in mouse neutrophils stimulated by lipopolysaccharide (LPS), an inflammatory inducer. Here, we show that bikunin: (i) blocks LPS-induced secretion of pro-inflammatory cytokines, including TNF-α and IL-1β, in a dose-dependent manner; (ii) has an inhibitory effect on cytokine production at a concentration of 0.2 µM, reaching 65% inhibition at the highest doses of bikunin tested (5 µM); (iii) has the suppressive capacity of ERK1/2 and p38 signaling pathways; and (iv) inhibited sequentially the LPS-induced phosphorylation of IκB-α, degradation of IκB-α, and nuclear translocation of NF-κB. When the MAPK data are analyzed, a significant decrease in phosphorylation is not seen at 0.2 µM bikunin but is at 1.0 µM dosing. Bikunin can inhibit LPS-induced neutrophil activation and cytokine release, although it is unlikely that it works primarily through the inhibition of MAPK phosphorylation. These data suggest that such effects are important in vivo and play a major contributory role in abrogation of neutrophil-mediated inflammatory responses, such as preterm delivery.

show abstract

Bikunin Target Genes in Ovarian Cancer Cells Identified by Microarray Analysis

Cited by 47 publications

References 45 publications

Transforming Growth Factor-β1-dependent Urokinase Up-regulation and Promotion of Invasion Are Involved in Src-MAPK-dependent Signaling in Human Ovarian Cancer Cells

Transforming Growth Factor-β1-dependent Urokinase Up-regulation and Promotion of Invasion Are Involved in Src-MAPK-dependent Signaling in Human Ovarian Cancer Cells

Reproductive failure in mice lacking inter-alpha-trypsin inhibitor (ITI) – ITI target genes in mouse ovary identified by microarray analysis

Bikunin suppresses expression of pro-inflammatory cytokines induced by lipopolysaccharide in neutrophils

Contact Info

Product

Resources

About