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The influence of monoacylglycerol lipase inhibition upon the expression of epidermal growth factor receptor in human PC-3 prostate cancer cells.

Cipriano M, Gouveia-Figueira S, Persson E, Nording M, Fowler CJ - BMC Res Notes (2014)

Bottom Line: CB1 receptor expression levels were found to differ greatly between two experimental series conducted using PC-3 cells.In the first series of experiments, JZL184 produced a small mitogenic effect for cells that had not been treated with EGF, whereas an anti-proliferative effect was seen for EGF-treated cells.An anti-proliferative effect for the EGF-treated cells was also seen with the CB receptor agonist CP55,940.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology and Clinical Neuroscience, Umeå University, Umeå, Sweden. cf@pharm.umu.se.

ABSTRACT

Background: It has been reported that direct activation of the cannabinoid CB1 receptor in epidermal growth factor (EGR)-stimulated PC-3 prostate cancer cells results in an anti-proliferative effect accompanied by a down-regulation of EGF receptors (EGFR). In the present study, we investigated whether similar effects are seen following inhibition of the endocannabinoid hydrolytic enzyme monoacylglycerol lipase (MGL).

Results: CB1 receptor expression levels were found to differ greatly between two experimental series conducted using PC-3 cells. The monoacylglycerol lipase inhibitor JZL184 increased levels of 2-arachidonoylglycerol in the PC-3 cells without producing changes in the levels of anandamide and related N-acylethanolamines. In the first series of experiments, JZL184 produced a small mitogenic effect for cells that had not been treated with EGF, whereas an anti-proliferative effect was seen for EGF-treated cells. An anti-proliferative effect for the EGF-treated cells was also seen with the CB receptor agonist CP55,940. In the second batch of cells, there was an interaction between JZL184 and CB1 receptor expression densities in linear regression analyses with EGFR expression as the dependent variable.

Conclusions: Inhibition of MGL by JZL184 can affect EGFR expression. However, the use in our hands of PC-3 cells as a model to investigate the therapeutic potential of MGL inhibitors and related compounds is compromised by their variability of CB1 receptor expression.

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Effects of JZL184 and CP55,940 upon PC-3 cell proliferation. Cells were incubated for 3 weeks in the absence or presence of EGF (10 ng/ml). Cells in 6 well plates were treated for three weeks without medium change in the absence or presence of 10 ng/ml of EGF. A and B: After two of the three weeks, either vehicle or JZL184 (1 μM) was added to the wells and the incubation was continued for a week without change of medium. In A, levels of 2-AG and N-acylethanolamines are shown. The data, taken for both PC-3 experimental series, show the total levels of the lipids in the cell extract expressed relative to the corresponding value for the extracts taken from wells containing control cells from the same plate. The bars show median values. Abbreviations: 2-AG, 2-arachidonoylglycerol; AEA, anandamide; PEA, palmitoylethanolamide; OEA, oleoylethanolamide; SEA, stearoylethanolamide. In B the data are from the first experimental series, and values are means ± s.e.m., n = 6. **P < 0.01, ***P < 0.001, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for JZL184. In C (first experimental series), the same experimental protocol was followed, but using 100 nM CP55,940 (“CP”). In C, data is given as means ± s.e.m., n = 7 (EGF-treated) or n = 3 (no EGF). **P < 0.01, NSnot significant, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for CP55,940.
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Figure 2: Effects of JZL184 and CP55,940 upon PC-3 cell proliferation. Cells were incubated for 3 weeks in the absence or presence of EGF (10 ng/ml). Cells in 6 well plates were treated for three weeks without medium change in the absence or presence of 10 ng/ml of EGF. A and B: After two of the three weeks, either vehicle or JZL184 (1 μM) was added to the wells and the incubation was continued for a week without change of medium. In A, levels of 2-AG and N-acylethanolamines are shown. The data, taken for both PC-3 experimental series, show the total levels of the lipids in the cell extract expressed relative to the corresponding value for the extracts taken from wells containing control cells from the same plate. The bars show median values. Abbreviations: 2-AG, 2-arachidonoylglycerol; AEA, anandamide; PEA, palmitoylethanolamide; OEA, oleoylethanolamide; SEA, stearoylethanolamide. In B the data are from the first experimental series, and values are means ± s.e.m., n = 6. **P < 0.01, ***P < 0.001, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for JZL184. In C (first experimental series), the same experimental protocol was followed, but using 100 nM CP55,940 (“CP”). In C, data is given as means ± s.e.m., n = 7 (EGF-treated) or n = 3 (no EGF). **P < 0.01, NSnot significant, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for CP55,940.

Mentions: As part of the first series of experiments with EGF, the cells were treated for the last week with either vehicle or JZL184 (1 μM). The JZL184 treatment produced the expected increase in 2-AG levels without affecting either AEA or related N-acylethanolamide levels (Figure 2A). The effect of this compound upon the cell proliferation is shown in Figure 2B. A two-way ANOVA matching for JZL184 indicated a significant interaction JZL184 × EGF (F1,10 = 84, P < 0.0001), due to a stimulation of cell proliferation by the compound for control cells and an inhibition for the EGF-treated cells.


The influence of monoacylglycerol lipase inhibition upon the expression of epidermal growth factor receptor in human PC-3 prostate cancer cells.

Cipriano M, Gouveia-Figueira S, Persson E, Nording M, Fowler CJ - BMC Res Notes (2014)

Effects of JZL184 and CP55,940 upon PC-3 cell proliferation. Cells were incubated for 3 weeks in the absence or presence of EGF (10 ng/ml). Cells in 6 well plates were treated for three weeks without medium change in the absence or presence of 10 ng/ml of EGF. A and B: After two of the three weeks, either vehicle or JZL184 (1 μM) was added to the wells and the incubation was continued for a week without change of medium. In A, levels of 2-AG and N-acylethanolamines are shown. The data, taken for both PC-3 experimental series, show the total levels of the lipids in the cell extract expressed relative to the corresponding value for the extracts taken from wells containing control cells from the same plate. The bars show median values. Abbreviations: 2-AG, 2-arachidonoylglycerol; AEA, anandamide; PEA, palmitoylethanolamide; OEA, oleoylethanolamide; SEA, stearoylethanolamide. In B the data are from the first experimental series, and values are means ± s.e.m., n = 6. **P < 0.01, ***P < 0.001, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for JZL184. In C (first experimental series), the same experimental protocol was followed, but using 100 nM CP55,940 (“CP”). In C, data is given as means ± s.e.m., n = 7 (EGF-treated) or n = 3 (no EGF). **P < 0.01, NSnot significant, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for CP55,940.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 2: Effects of JZL184 and CP55,940 upon PC-3 cell proliferation. Cells were incubated for 3 weeks in the absence or presence of EGF (10 ng/ml). Cells in 6 well plates were treated for three weeks without medium change in the absence or presence of 10 ng/ml of EGF. A and B: After two of the three weeks, either vehicle or JZL184 (1 μM) was added to the wells and the incubation was continued for a week without change of medium. In A, levels of 2-AG and N-acylethanolamines are shown. The data, taken for both PC-3 experimental series, show the total levels of the lipids in the cell extract expressed relative to the corresponding value for the extracts taken from wells containing control cells from the same plate. The bars show median values. Abbreviations: 2-AG, 2-arachidonoylglycerol; AEA, anandamide; PEA, palmitoylethanolamide; OEA, oleoylethanolamide; SEA, stearoylethanolamide. In B the data are from the first experimental series, and values are means ± s.e.m., n = 6. **P < 0.01, ***P < 0.001, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for JZL184. In C (first experimental series), the same experimental protocol was followed, but using 100 nM CP55,940 (“CP”). In C, data is given as means ± s.e.m., n = 7 (EGF-treated) or n = 3 (no EGF). **P < 0.01, NSnot significant, Šídák’s multiple comparison test vs. the corresponding vehicle control following a significant interaction term in the two-way ANOVA matching for CP55,940.
Mentions: As part of the first series of experiments with EGF, the cells were treated for the last week with either vehicle or JZL184 (1 μM). The JZL184 treatment produced the expected increase in 2-AG levels without affecting either AEA or related N-acylethanolamide levels (Figure 2A). The effect of this compound upon the cell proliferation is shown in Figure 2B. A two-way ANOVA matching for JZL184 indicated a significant interaction JZL184 × EGF (F1,10 = 84, P < 0.0001), due to a stimulation of cell proliferation by the compound for control cells and an inhibition for the EGF-treated cells.

Bottom Line: CB1 receptor expression levels were found to differ greatly between two experimental series conducted using PC-3 cells.In the first series of experiments, JZL184 produced a small mitogenic effect for cells that had not been treated with EGF, whereas an anti-proliferative effect was seen for EGF-treated cells.An anti-proliferative effect for the EGF-treated cells was also seen with the CB receptor agonist CP55,940.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology and Clinical Neuroscience, Umeå University, Umeå, Sweden. cf@pharm.umu.se.

ABSTRACT

Background: It has been reported that direct activation of the cannabinoid CB1 receptor in epidermal growth factor (EGR)-stimulated PC-3 prostate cancer cells results in an anti-proliferative effect accompanied by a down-regulation of EGF receptors (EGFR). In the present study, we investigated whether similar effects are seen following inhibition of the endocannabinoid hydrolytic enzyme monoacylglycerol lipase (MGL).

Results: CB1 receptor expression levels were found to differ greatly between two experimental series conducted using PC-3 cells. The monoacylglycerol lipase inhibitor JZL184 increased levels of 2-arachidonoylglycerol in the PC-3 cells without producing changes in the levels of anandamide and related N-acylethanolamines. In the first series of experiments, JZL184 produced a small mitogenic effect for cells that had not been treated with EGF, whereas an anti-proliferative effect was seen for EGF-treated cells. An anti-proliferative effect for the EGF-treated cells was also seen with the CB receptor agonist CP55,940. In the second batch of cells, there was an interaction between JZL184 and CB1 receptor expression densities in linear regression analyses with EGFR expression as the dependent variable.

Conclusions: Inhibition of MGL by JZL184 can affect EGFR expression. However, the use in our hands of PC-3 cells as a model to investigate the therapeutic potential of MGL inhibitors and related compounds is compromised by their variability of CB1 receptor expression.

Show MeSH
Related in: MedlinePlus