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Lipoxin A4 Attenuates Cell Invasion by Inhibiting ROS/ERK/MMP Pathway in Pancreatic Cancer.

Zong L, Li J, Chen X, Chen K, Li W, Li X, Zhang L, Duan W, Lei J, Xu Q, Shan T, Ma Q, Sun H - Oxid Med Cell Longev (2015)

Bottom Line: Our data showed that LXA4 significantly inhibited both cell invasion and the expression of matrix metalloproteinase- (MMP-) 9 and MMP-2.Further experiments implied that LXA4 decreased the levels of intracellular reactive oxygen species (ROS) and the activity of the extracellular signal regulated kinases (ERK) pathway to achieve similar outcome to ROS scavenger N-acetyl-L-cysteine (NAC).However, a decreased level of intracellular ROS was not observed in cells treated with the specific ERK pathway inhibitor FR180204.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China.

ABSTRACT
Lipoxin A4 (LXA4), an endogenous arachidonic acid metabolite, was previously considered an anti-inflammatory lipid mediator. But it also has the potential to inhibit cancer progression. To explore the therapeutic effect of LXA4 in pancreatic cancer, we used Panc-1 cells to investigate the mechanism by which LXA4 can attenuate pancreatic cancer cell invasion. Our data showed that LXA4 significantly inhibited both cell invasion and the expression of matrix metalloproteinase- (MMP-) 9 and MMP-2. Further experiments implied that LXA4 decreased the levels of intracellular reactive oxygen species (ROS) and the activity of the extracellular signal regulated kinases (ERK) pathway to achieve similar outcome to ROS scavenger N-acetyl-L-cysteine (NAC). However, a decreased level of intracellular ROS was not observed in cells treated with the specific ERK pathway inhibitor FR180204. The blocking of either intracellular ROS or ERK pathway caused the downregulation of MMP-9 and MMP-2 expression. Furthermore, tests revealed that LXA4 inhibited MMP-9 and MMP-2 at the mRNA, protein, and functional levels. Finally, LXA4 dramatically limited the invasion of CoCl2-mimic hypoxic cells and abrogated intracellular ROS levels, ERK activity, and MMPs expression. These results suggest that LXA4 attenuates cell invasion in pancreatic cancer by suppressing the ROS/ERK/MMPs pathway, which may be beneficial for preventing the invasion of pancreatic cancer.

No MeSH data available.


Related in: MedlinePlus

LXA4 reverses CoCl2-induced cell invasion through ROS/ERK/MMP pathway. (a) Effect of LXA4 on CoCl2-induced cell invasion. Panc-1 cells were treated with vehicle (methanol), LXA4, CoCl2 (0.15 mM), or CoCl2 + LXA4. Cell invasion assay was performed when cells had been transferred into transwell chamber for 48 hours. (b) The quantified data of (a). (c) Western blot analysis of cells treated as above. (d) Intracellular ROS determined in cells treated in (a). Cells incubated with DCF-DA for 20 min were washed with PBS three times and then lysed by RIPA lysis buffer and tested by fluorimetry at 510 nm. The absorbance was normalized by total protein. (e) Expression of activated p-ERK and total ERK detected by western blot.  ∗P < 0.05 versus corresponding control.
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fig5: LXA4 reverses CoCl2-induced cell invasion through ROS/ERK/MMP pathway. (a) Effect of LXA4 on CoCl2-induced cell invasion. Panc-1 cells were treated with vehicle (methanol), LXA4, CoCl2 (0.15 mM), or CoCl2 + LXA4. Cell invasion assay was performed when cells had been transferred into transwell chamber for 48 hours. (b) The quantified data of (a). (c) Western blot analysis of cells treated as above. (d) Intracellular ROS determined in cells treated in (a). Cells incubated with DCF-DA for 20 min were washed with PBS three times and then lysed by RIPA lysis buffer and tested by fluorimetry at 510 nm. The absorbance was normalized by total protein. (e) Expression of activated p-ERK and total ERK detected by western blot.  ∗P < 0.05 versus corresponding control.

Mentions: According to our previous study [21, 22], pancreatic cancer is a type of malignancy that demonstrates poor perfusion, and consequently a hypoxic microenvironment can dramatically increase intracellular ROS which may promote cell invasion and epithelial-mesenchymal transition (EMT). To test whether hypoxia could increase MMP-9 and MMP-2 levels and whether LXA4 could reverse this overexpression, we added 0.15 mM CoCl2 to mimic the cellular hypoxic state. In cell invasion assay, after a comparison with cells that were treated with vehicle control, we found that cells treated with CoCl2 became more aggressive in nature. However, when they were treated with CoCl2 + LXA4, the number of cells that passed through the Matrigel decreased (Figures 5(a) and 5(b)), which suggested LXA4 reversed CoCl2-induced cell invasion. Next, the expression of MMP was measured. Cells that were treated with CoCl2 overexpressed MMP-9 and MMP-2, which was reversed by CoCl2 + LXA4 (Figure 5(c)). This demonstrates that LXA4 could reverse the CoCl2-induced overexpression of MMPs. Furthermore, an assay to determine intracellular ROS assay showed that CoCl2 upregulated intracellular ROS while LXA4 could attenuate that effect (Figure 5(d)). In addition, the cellular ERK pathway was activated when the cells were cultured with CoCl2, but it was inactivated by LXA4 (Figure 5(e)). These data implied that inactivation of the ROS/ERK/MMP pathway might be involved in the reversal of CoCl2-induced cell invasion.


Lipoxin A4 Attenuates Cell Invasion by Inhibiting ROS/ERK/MMP Pathway in Pancreatic Cancer.

Zong L, Li J, Chen X, Chen K, Li W, Li X, Zhang L, Duan W, Lei J, Xu Q, Shan T, Ma Q, Sun H - Oxid Med Cell Longev (2015)

LXA4 reverses CoCl2-induced cell invasion through ROS/ERK/MMP pathway. (a) Effect of LXA4 on CoCl2-induced cell invasion. Panc-1 cells were treated with vehicle (methanol), LXA4, CoCl2 (0.15 mM), or CoCl2 + LXA4. Cell invasion assay was performed when cells had been transferred into transwell chamber for 48 hours. (b) The quantified data of (a). (c) Western blot analysis of cells treated as above. (d) Intracellular ROS determined in cells treated in (a). Cells incubated with DCF-DA for 20 min were washed with PBS three times and then lysed by RIPA lysis buffer and tested by fluorimetry at 510 nm. The absorbance was normalized by total protein. (e) Expression of activated p-ERK and total ERK detected by western blot.  ∗P < 0.05 versus corresponding control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4663743&req=5

fig5: LXA4 reverses CoCl2-induced cell invasion through ROS/ERK/MMP pathway. (a) Effect of LXA4 on CoCl2-induced cell invasion. Panc-1 cells were treated with vehicle (methanol), LXA4, CoCl2 (0.15 mM), or CoCl2 + LXA4. Cell invasion assay was performed when cells had been transferred into transwell chamber for 48 hours. (b) The quantified data of (a). (c) Western blot analysis of cells treated as above. (d) Intracellular ROS determined in cells treated in (a). Cells incubated with DCF-DA for 20 min were washed with PBS three times and then lysed by RIPA lysis buffer and tested by fluorimetry at 510 nm. The absorbance was normalized by total protein. (e) Expression of activated p-ERK and total ERK detected by western blot.  ∗P < 0.05 versus corresponding control.
Mentions: According to our previous study [21, 22], pancreatic cancer is a type of malignancy that demonstrates poor perfusion, and consequently a hypoxic microenvironment can dramatically increase intracellular ROS which may promote cell invasion and epithelial-mesenchymal transition (EMT). To test whether hypoxia could increase MMP-9 and MMP-2 levels and whether LXA4 could reverse this overexpression, we added 0.15 mM CoCl2 to mimic the cellular hypoxic state. In cell invasion assay, after a comparison with cells that were treated with vehicle control, we found that cells treated with CoCl2 became more aggressive in nature. However, when they were treated with CoCl2 + LXA4, the number of cells that passed through the Matrigel decreased (Figures 5(a) and 5(b)), which suggested LXA4 reversed CoCl2-induced cell invasion. Next, the expression of MMP was measured. Cells that were treated with CoCl2 overexpressed MMP-9 and MMP-2, which was reversed by CoCl2 + LXA4 (Figure 5(c)). This demonstrates that LXA4 could reverse the CoCl2-induced overexpression of MMPs. Furthermore, an assay to determine intracellular ROS assay showed that CoCl2 upregulated intracellular ROS while LXA4 could attenuate that effect (Figure 5(d)). In addition, the cellular ERK pathway was activated when the cells were cultured with CoCl2, but it was inactivated by LXA4 (Figure 5(e)). These data implied that inactivation of the ROS/ERK/MMP pathway might be involved in the reversal of CoCl2-induced cell invasion.

Bottom Line: Our data showed that LXA4 significantly inhibited both cell invasion and the expression of matrix metalloproteinase- (MMP-) 9 and MMP-2.Further experiments implied that LXA4 decreased the levels of intracellular reactive oxygen species (ROS) and the activity of the extracellular signal regulated kinases (ERK) pathway to achieve similar outcome to ROS scavenger N-acetyl-L-cysteine (NAC).However, a decreased level of intracellular ROS was not observed in cells treated with the specific ERK pathway inhibitor FR180204.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China.

ABSTRACT
Lipoxin A4 (LXA4), an endogenous arachidonic acid metabolite, was previously considered an anti-inflammatory lipid mediator. But it also has the potential to inhibit cancer progression. To explore the therapeutic effect of LXA4 in pancreatic cancer, we used Panc-1 cells to investigate the mechanism by which LXA4 can attenuate pancreatic cancer cell invasion. Our data showed that LXA4 significantly inhibited both cell invasion and the expression of matrix metalloproteinase- (MMP-) 9 and MMP-2. Further experiments implied that LXA4 decreased the levels of intracellular reactive oxygen species (ROS) and the activity of the extracellular signal regulated kinases (ERK) pathway to achieve similar outcome to ROS scavenger N-acetyl-L-cysteine (NAC). However, a decreased level of intracellular ROS was not observed in cells treated with the specific ERK pathway inhibitor FR180204. The blocking of either intracellular ROS or ERK pathway caused the downregulation of MMP-9 and MMP-2 expression. Furthermore, tests revealed that LXA4 inhibited MMP-9 and MMP-2 at the mRNA, protein, and functional levels. Finally, LXA4 dramatically limited the invasion of CoCl2-mimic hypoxic cells and abrogated intracellular ROS levels, ERK activity, and MMPs expression. These results suggest that LXA4 attenuates cell invasion in pancreatic cancer by suppressing the ROS/ERK/MMPs pathway, which may be beneficial for preventing the invasion of pancreatic cancer.

No MeSH data available.


Related in: MedlinePlus