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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 negatively regulated cell invasion by inhibiting ROS/ERK pathway. (a) Influence of LXA4 on cell invasion in Panc-1 cells. Cells were treated with vehicle (methanol), LXA4 (400 nM), ROS scavenger NAC (20 mM), or ERK specific inhibitor FR180204 (10 μM) for 24 hours. Then 1 × 105 cells were transferred into transwell chambers covered with Matrigel. After forty-eight hours, cells were stained with 0.1% crystal violet, observed, and counted under microscope. (b) The quantified results of (a). (c) Representative western blot analysis of activated p-ERK and total ERK in cells treated as in (a). (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. It was normalized by total protein.  ∗P < 0.05 versus vehicle control.
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fig3: LXA4 negatively regulated cell invasion by inhibiting ROS/ERK pathway. (a) Influence of LXA4 on cell invasion in Panc-1 cells. Cells were treated with vehicle (methanol), LXA4 (400 nM), ROS scavenger NAC (20 mM), or ERK specific inhibitor FR180204 (10 μM) for 24 hours. Then 1 × 105 cells were transferred into transwell chambers covered with Matrigel. After forty-eight hours, cells were stained with 0.1% crystal violet, observed, and counted under microscope. (b) The quantified results of (a). (c) Representative western blot analysis of activated p-ERK and total ERK in cells treated as in (a). (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. It was normalized by total protein.  ∗P < 0.05 versus vehicle control.

Mentions: The ERK pathway, which is overactive in pancreatic cancer, is widely accepted to affect cell invasion [20]. When exposed to the specific ERK pathway inhibitor FR180204 (10 μM), cells present less aggressive invasion as LXA4 and NAC (Figures 3(a) and 3(b)), which suggests that ERK might mediate LXA4 attenuated cell invasion. Because ERK is reported to be a downstream pathway of ROS [12, 13], we detected ERK activity and showed phospho-ERK accounted for a lower proportion of total ERK when the cells were treated with LXA4 and NAC (Figure 3(c)). However, cells that were exposed to FR180204 failed to show a decrease in intracellular ROS (Figure 3(d)). Our data confirmed that ROS could induce ERK activation, which suggests that LXA4 could inactivate the ERK pathway via decreasing intracellular ROS. This in turn further downregulates 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 negatively regulated cell invasion by inhibiting ROS/ERK pathway. (a) Influence of LXA4 on cell invasion in Panc-1 cells. Cells were treated with vehicle (methanol), LXA4 (400 nM), ROS scavenger NAC (20 mM), or ERK specific inhibitor FR180204 (10 μM) for 24 hours. Then 1 × 105 cells were transferred into transwell chambers covered with Matrigel. After forty-eight hours, cells were stained with 0.1% crystal violet, observed, and counted under microscope. (b) The quantified results of (a). (c) Representative western blot analysis of activated p-ERK and total ERK in cells treated as in (a). (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. It was normalized by total protein.  ∗P < 0.05 versus vehicle control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: LXA4 negatively regulated cell invasion by inhibiting ROS/ERK pathway. (a) Influence of LXA4 on cell invasion in Panc-1 cells. Cells were treated with vehicle (methanol), LXA4 (400 nM), ROS scavenger NAC (20 mM), or ERK specific inhibitor FR180204 (10 μM) for 24 hours. Then 1 × 105 cells were transferred into transwell chambers covered with Matrigel. After forty-eight hours, cells were stained with 0.1% crystal violet, observed, and counted under microscope. (b) The quantified results of (a). (c) Representative western blot analysis of activated p-ERK and total ERK in cells treated as in (a). (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. It was normalized by total protein.  ∗P < 0.05 versus vehicle control.
Mentions: The ERK pathway, which is overactive in pancreatic cancer, is widely accepted to affect cell invasion [20]. When exposed to the specific ERK pathway inhibitor FR180204 (10 μM), cells present less aggressive invasion as LXA4 and NAC (Figures 3(a) and 3(b)), which suggests that ERK might mediate LXA4 attenuated cell invasion. Because ERK is reported to be a downstream pathway of ROS [12, 13], we detected ERK activity and showed phospho-ERK accounted for a lower proportion of total ERK when the cells were treated with LXA4 and NAC (Figure 3(c)). However, cells that were exposed to FR180204 failed to show a decrease in intracellular ROS (Figure 3(d)). Our data confirmed that ROS could induce ERK activation, which suggests that LXA4 could inactivate the ERK pathway via decreasing intracellular ROS. This in turn further downregulates 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