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Reactive oxygen species regulate urokinase plasminogen activator expression and cell invasion via mitogen-activated protein kinase pathways after treatment with hepatocyte growth factor in stomach cancer cells.

Lee KH, Kim SW, Kim JR - J. Exp. Clin. Cancer Res. (2009)

Bottom Line: We confirmed that Rac-1 regulated ROS production after activation of the AKT pathway with HGF.Exogenously added H2O2 promoted the expression of HGF, but not in a dose-dependent manner and also showed negative expression of HGF after co-treatment with H2O2 and HGF.We clarified the downstream pathways regulated by ROS after treatment with H2O2, which showed negative control between FRK and p38 kinase activities for uPA regulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Biochemistry and Molecular Biology, College of Medicine, Yeungnam University, Daegu, Korea. lkhee@med.yu.ac.kr

ABSTRACT

Background: Reactive oxygen species (ROS) are closely associated with the intracellular signal cascade, thus strongly implicating involvement in tumor progression. However, the mechanism by which ROS are generated and how ROS target downstream molecules to trigger tumor metastasis is unclear. In this study, we investigated the underlying signal pathways in ROS-induced urokinase plasminogen activator (uPA) expression in the human gastric cancer cells, NUGC-3 and MKN-28.

Methods and results: Intracellular ROS, as determined using the fluorescent probe, 2'-7' dichlorofluorescein diacetate, decreased after treatment with hepatocyte growth factor (HGF). We confirmed that Rac-1 regulated ROS production after activation of the AKT pathway with HGF. Exogenously added H2O2 promoted the expression of HGF, but not in a dose-dependent manner and also showed negative expression of HGF after co-treatment with H2O2 and HGF. Treatment with NAC, an intracellular free radical scavenger, decreased the enhancement of uPA production and tumor invasion in both cells. We clarified the downstream pathways regulated by ROS after treatment with H2O2, which showed negative control between FRK and p38 kinase activities for uPA regulation.

Conclusion: HGF regulates Rac-1-induced ROS production through the Akt pathway and ROS regulates uPA production and invasion via MAP kinase, which provides novel insight into the mechanisms underlying the progression of gastric cancer.

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Level of expression of HGF after treatment with H2O2 and/or HGF. Cells were serum-starved and treated with H2O2 (100 μM) and/or HGF (10 ng/ml). The level of HGF mRNA was measured by real-time RT-PCR analysis. Values are the means ± SD of triplicates of three independent experiments. Statistical significance was estimated by Student's t-test (*, p < 0.05;**, p < 0.01).
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Figure 7: Level of expression of HGF after treatment with H2O2 and/or HGF. Cells were serum-starved and treated with H2O2 (100 μM) and/or HGF (10 ng/ml). The level of HGF mRNA was measured by real-time RT-PCR analysis. Values are the means ± SD of triplicates of three independent experiments. Statistical significance was estimated by Student's t-test (*, p < 0.05;**, p < 0.01).

Mentions: To understand the mechanism of uPA production of ROS, we examined HGF gene expression using the RT-PCR method. The levels of HGF mRNA were 1.7–2.4 fold higher in cells treated with 100 μM H2O2 than in untreated cells. However, HGF mRNA levels were decreased when treated with 500 μM H2O2 (Figure 6). This might be due to H2O2 cytotoxicity. Subsequently, we measured HGF mRNA levels from both cell lines in the absence or presence of exogenous HGF and/or H2O2. The levels of HGF mRNA were suppressed by exogenous treatment of HGF and H2O2 (Figure 7).


Reactive oxygen species regulate urokinase plasminogen activator expression and cell invasion via mitogen-activated protein kinase pathways after treatment with hepatocyte growth factor in stomach cancer cells.

Lee KH, Kim SW, Kim JR - J. Exp. Clin. Cancer Res. (2009)

Level of expression of HGF after treatment with H2O2 and/or HGF. Cells were serum-starved and treated with H2O2 (100 μM) and/or HGF (10 ng/ml). The level of HGF mRNA was measured by real-time RT-PCR analysis. Values are the means ± SD of triplicates of three independent experiments. Statistical significance was estimated by Student's t-test (*, p < 0.05;**, p < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2698863&req=5

Figure 7: Level of expression of HGF after treatment with H2O2 and/or HGF. Cells were serum-starved and treated with H2O2 (100 μM) and/or HGF (10 ng/ml). The level of HGF mRNA was measured by real-time RT-PCR analysis. Values are the means ± SD of triplicates of three independent experiments. Statistical significance was estimated by Student's t-test (*, p < 0.05;**, p < 0.01).
Mentions: To understand the mechanism of uPA production of ROS, we examined HGF gene expression using the RT-PCR method. The levels of HGF mRNA were 1.7–2.4 fold higher in cells treated with 100 μM H2O2 than in untreated cells. However, HGF mRNA levels were decreased when treated with 500 μM H2O2 (Figure 6). This might be due to H2O2 cytotoxicity. Subsequently, we measured HGF mRNA levels from both cell lines in the absence or presence of exogenous HGF and/or H2O2. The levels of HGF mRNA were suppressed by exogenous treatment of HGF and H2O2 (Figure 7).

Bottom Line: We confirmed that Rac-1 regulated ROS production after activation of the AKT pathway with HGF.Exogenously added H2O2 promoted the expression of HGF, but not in a dose-dependent manner and also showed negative expression of HGF after co-treatment with H2O2 and HGF.We clarified the downstream pathways regulated by ROS after treatment with H2O2, which showed negative control between FRK and p38 kinase activities for uPA regulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Biochemistry and Molecular Biology, College of Medicine, Yeungnam University, Daegu, Korea. lkhee@med.yu.ac.kr

ABSTRACT

Background: Reactive oxygen species (ROS) are closely associated with the intracellular signal cascade, thus strongly implicating involvement in tumor progression. However, the mechanism by which ROS are generated and how ROS target downstream molecules to trigger tumor metastasis is unclear. In this study, we investigated the underlying signal pathways in ROS-induced urokinase plasminogen activator (uPA) expression in the human gastric cancer cells, NUGC-3 and MKN-28.

Methods and results: Intracellular ROS, as determined using the fluorescent probe, 2'-7' dichlorofluorescein diacetate, decreased after treatment with hepatocyte growth factor (HGF). We confirmed that Rac-1 regulated ROS production after activation of the AKT pathway with HGF. Exogenously added H2O2 promoted the expression of HGF, but not in a dose-dependent manner and also showed negative expression of HGF after co-treatment with H2O2 and HGF. Treatment with NAC, an intracellular free radical scavenger, decreased the enhancement of uPA production and tumor invasion in both cells. We clarified the downstream pathways regulated by ROS after treatment with H2O2, which showed negative control between FRK and p38 kinase activities for uPA regulation.

Conclusion: HGF regulates Rac-1-induced ROS production through the Akt pathway and ROS regulates uPA production and invasion via MAP kinase, which provides novel insight into the mechanisms underlying the progression of gastric cancer.

Show MeSH
Related in: MedlinePlus