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Decursin and decursinol angelate inhibit estrogen-stimulated and estrogen-independent growth and survival of breast cancer cells.

Jiang C, Guo J, Wang Z, Xiao B, Lee HJ, Lee EO, Kim SH, Lu J - Breast Cancer Res. (2007)

Bottom Line: Decursin and DA exerted growth inhibitory effects on MCF-7 cells through G1 arrest and caspase-mediated apoptosis.Decursin and the pure antiestrogen Faslodex exerted an additive growth inhibitory effect on MCF-7 cells.In contrast, decursinol, which lacks the side chain of decursin and DA, did not have these cellular and molecular activities at comparable concentrations.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Hormel Institute, University of Minnesota, 801 16th Avenue NE, Austin, MN 55912, USA.

ABSTRACT

Introduction: Estrogen and estrogen receptor (ER)-mediated signaling are crucial for the etiology and progression of human breast cancer. Attenuating ER activities by natural products is a promising strategy to decrease breast cancer risk. We recently discovered that the pyranocoumarin compound decursin and its isomer decursinol angelate (DA) have potent novel antiandrogen receptor signaling activities. Because the ER and the androgen receptor belong to the steroid receptor superfamily, we examined whether these compounds affected ER expression and signaling in breast cancer cells.

Methods: We treated estrogen-dependent MCF-7 and estrogen-independent MDA MB-231 human breast cancer cells with decursin and DA, and examined cell growth, apoptosis, and ERalpha and ERbeta expression in both cell lines - and, in particular, estrogen-stimulated signaling in the MCF-7 cells. We compared these compounds with decursinol to determine their structure-activity relationship.

Results: Decursin and DA exerted growth inhibitory effects on MCF-7 cells through G1 arrest and caspase-mediated apoptosis. These compounds decreased ERalpha in MCF-7 cells at both mRNA and protein levels, and suppressed estrogen-stimulated genes. Decursin and the pure antiestrogen Faslodex exerted an additive growth inhibitory effect on MCF-7 cells. In MDA MB-231 cells, these compounds induced cell-cycle arrests in the G1 and G2 phases as well as inducing apoptosis, accompanied by an increased expression of ERbeta. In contrast, decursinol, which lacks the side chain of decursin and DA, did not have these cellular and molecular activities at comparable concentrations.

Conclusion: The side chain of decursin and DA is crucial for their anti-ER signaling and breast cancer growth inhibitory activities. These data provide mechanistic rationales for validating the chemopreventive and therapeutic efficacy of decursin and its derivatives in preclinical animal models of breast cancer.

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Effects of pyranocoumarins on estrogen receptor alpha nuclear translocation and transactivation in MCF-7 cells. (a), (b) Inhibitory effects of decursin on estrogen receptor (ER)α nuclear translocation in MCF-7 cells. (a) The time course of ERα nuclear translocation in MCF-7 cells stimulated by 17β-estradiol (E2) was first established in estrogen-starved MCF-7 cells. Fresh medium without or with 1 nM E2 was added. At different time points, nuclear (N) and cytosolic (C) fractions were prepared for western blot analyses. Poly(ADP-ribose) polymerase (PARP) and α-tubulin were detected as markers of N and C proteins, respectively. DMSO, dimethylsulfoxide. (b) For the decursin experiment, estrogen-deprived MCF-7 cells were pretreated with decursin for 2 hours and were stimulated with 1 nM E2 for 1 hour (total decursin exposure time, 3 hours). N and C fractions were prepared for western blot analyses. One-tenth nuclear fraction was used compared with the time-course experiment to show the difference. The total lysate ERα level was determined (right panel). (c) Differential effects of decursin versus decursinol on estrogen-response element (ERE) activity in MCF-7 cells: left, with E2; right, without, E2. An aliquot of 1 × 105 MCF-7 cells was placed in a 12-well plate and cotransfected with ERE-luciferase reporter plasmid and pSV-β-galactosidase reporter in phenol-red free improved minimum essential medium (PRF-IMEM) without serum and insulin. After transfection for 24 hours, the cells were treated with different concentrations of decursin (D, 20 μM, 50 μM), decursinol (DL, 100 μM) or DMSO in the absence or presence of 1 nM E2 for 24 hours. Cells extracts were prepared for luciferase activity and β-galactosidase activity. Mean ± standard error, three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus basal or E2-stimulated control.
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Figure 6: Effects of pyranocoumarins on estrogen receptor alpha nuclear translocation and transactivation in MCF-7 cells. (a), (b) Inhibitory effects of decursin on estrogen receptor (ER)α nuclear translocation in MCF-7 cells. (a) The time course of ERα nuclear translocation in MCF-7 cells stimulated by 17β-estradiol (E2) was first established in estrogen-starved MCF-7 cells. Fresh medium without or with 1 nM E2 was added. At different time points, nuclear (N) and cytosolic (C) fractions were prepared for western blot analyses. Poly(ADP-ribose) polymerase (PARP) and α-tubulin were detected as markers of N and C proteins, respectively. DMSO, dimethylsulfoxide. (b) For the decursin experiment, estrogen-deprived MCF-7 cells were pretreated with decursin for 2 hours and were stimulated with 1 nM E2 for 1 hour (total decursin exposure time, 3 hours). N and C fractions were prepared for western blot analyses. One-tenth nuclear fraction was used compared with the time-course experiment to show the difference. The total lysate ERα level was determined (right panel). (c) Differential effects of decursin versus decursinol on estrogen-response element (ERE) activity in MCF-7 cells: left, with E2; right, without, E2. An aliquot of 1 × 105 MCF-7 cells was placed in a 12-well plate and cotransfected with ERE-luciferase reporter plasmid and pSV-β-galactosidase reporter in phenol-red free improved minimum essential medium (PRF-IMEM) without serum and insulin. After transfection for 24 hours, the cells were treated with different concentrations of decursin (D, 20 μM, 50 μM), decursinol (DL, 100 μM) or DMSO in the absence or presence of 1 nM E2 for 24 hours. Cells extracts were prepared for luciferase activity and β-galactosidase activity. Mean ± standard error, three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus basal or E2-stimulated control.

Mentions: Because decursin and DA potently inhibit the cytosol to nuclear translocation of AR upon androgen stimulation in prostate cancer cells [26,27], we investigated whether a similar inhibitory effect was exerted on ERα. We first determined the optimal timeframe to study ERα nuclear translocation in MCF-7 cells. As shown in Figure 6a (lane 2 versus lane 1), the ERα was predominantly localized in the nucleus in MCF-7 cells grown in PRF-IMEM medium without serum and insulin for 48 hours. This was in contrast to the predominant cytosolic distribution of AR in androgen-deprived prostate cancer cells [26]. Nevertheless, a decrease of cytosolic ERα (Figure 6a, lane 3 versus lane 1) and a reciprocal increase of nuclear ERα (Figure 6a, lane 4 versus lane 2) could be detected as soon as 30 minutes of stimulation by 17β-estradiol, and the changes persisted through 4 hours of stimulation (Figure 4a).


Decursin and decursinol angelate inhibit estrogen-stimulated and estrogen-independent growth and survival of breast cancer cells.

Jiang C, Guo J, Wang Z, Xiao B, Lee HJ, Lee EO, Kim SH, Lu J - Breast Cancer Res. (2007)

Effects of pyranocoumarins on estrogen receptor alpha nuclear translocation and transactivation in MCF-7 cells. (a), (b) Inhibitory effects of decursin on estrogen receptor (ER)α nuclear translocation in MCF-7 cells. (a) The time course of ERα nuclear translocation in MCF-7 cells stimulated by 17β-estradiol (E2) was first established in estrogen-starved MCF-7 cells. Fresh medium without or with 1 nM E2 was added. At different time points, nuclear (N) and cytosolic (C) fractions were prepared for western blot analyses. Poly(ADP-ribose) polymerase (PARP) and α-tubulin were detected as markers of N and C proteins, respectively. DMSO, dimethylsulfoxide. (b) For the decursin experiment, estrogen-deprived MCF-7 cells were pretreated with decursin for 2 hours and were stimulated with 1 nM E2 for 1 hour (total decursin exposure time, 3 hours). N and C fractions were prepared for western blot analyses. One-tenth nuclear fraction was used compared with the time-course experiment to show the difference. The total lysate ERα level was determined (right panel). (c) Differential effects of decursin versus decursinol on estrogen-response element (ERE) activity in MCF-7 cells: left, with E2; right, without, E2. An aliquot of 1 × 105 MCF-7 cells was placed in a 12-well plate and cotransfected with ERE-luciferase reporter plasmid and pSV-β-galactosidase reporter in phenol-red free improved minimum essential medium (PRF-IMEM) without serum and insulin. After transfection for 24 hours, the cells were treated with different concentrations of decursin (D, 20 μM, 50 μM), decursinol (DL, 100 μM) or DMSO in the absence or presence of 1 nM E2 for 24 hours. Cells extracts were prepared for luciferase activity and β-galactosidase activity. Mean ± standard error, three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus basal or E2-stimulated control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2246173&req=5

Figure 6: Effects of pyranocoumarins on estrogen receptor alpha nuclear translocation and transactivation in MCF-7 cells. (a), (b) Inhibitory effects of decursin on estrogen receptor (ER)α nuclear translocation in MCF-7 cells. (a) The time course of ERα nuclear translocation in MCF-7 cells stimulated by 17β-estradiol (E2) was first established in estrogen-starved MCF-7 cells. Fresh medium without or with 1 nM E2 was added. At different time points, nuclear (N) and cytosolic (C) fractions were prepared for western blot analyses. Poly(ADP-ribose) polymerase (PARP) and α-tubulin were detected as markers of N and C proteins, respectively. DMSO, dimethylsulfoxide. (b) For the decursin experiment, estrogen-deprived MCF-7 cells were pretreated with decursin for 2 hours and were stimulated with 1 nM E2 for 1 hour (total decursin exposure time, 3 hours). N and C fractions were prepared for western blot analyses. One-tenth nuclear fraction was used compared with the time-course experiment to show the difference. The total lysate ERα level was determined (right panel). (c) Differential effects of decursin versus decursinol on estrogen-response element (ERE) activity in MCF-7 cells: left, with E2; right, without, E2. An aliquot of 1 × 105 MCF-7 cells was placed in a 12-well plate and cotransfected with ERE-luciferase reporter plasmid and pSV-β-galactosidase reporter in phenol-red free improved minimum essential medium (PRF-IMEM) without serum and insulin. After transfection for 24 hours, the cells were treated with different concentrations of decursin (D, 20 μM, 50 μM), decursinol (DL, 100 μM) or DMSO in the absence or presence of 1 nM E2 for 24 hours. Cells extracts were prepared for luciferase activity and β-galactosidase activity. Mean ± standard error, three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus basal or E2-stimulated control.
Mentions: Because decursin and DA potently inhibit the cytosol to nuclear translocation of AR upon androgen stimulation in prostate cancer cells [26,27], we investigated whether a similar inhibitory effect was exerted on ERα. We first determined the optimal timeframe to study ERα nuclear translocation in MCF-7 cells. As shown in Figure 6a (lane 2 versus lane 1), the ERα was predominantly localized in the nucleus in MCF-7 cells grown in PRF-IMEM medium without serum and insulin for 48 hours. This was in contrast to the predominant cytosolic distribution of AR in androgen-deprived prostate cancer cells [26]. Nevertheless, a decrease of cytosolic ERα (Figure 6a, lane 3 versus lane 1) and a reciprocal increase of nuclear ERα (Figure 6a, lane 4 versus lane 2) could be detected as soon as 30 minutes of stimulation by 17β-estradiol, and the changes persisted through 4 hours of stimulation (Figure 4a).

Bottom Line: Decursin and DA exerted growth inhibitory effects on MCF-7 cells through G1 arrest and caspase-mediated apoptosis.Decursin and the pure antiestrogen Faslodex exerted an additive growth inhibitory effect on MCF-7 cells.In contrast, decursinol, which lacks the side chain of decursin and DA, did not have these cellular and molecular activities at comparable concentrations.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Hormel Institute, University of Minnesota, 801 16th Avenue NE, Austin, MN 55912, USA.

ABSTRACT

Introduction: Estrogen and estrogen receptor (ER)-mediated signaling are crucial for the etiology and progression of human breast cancer. Attenuating ER activities by natural products is a promising strategy to decrease breast cancer risk. We recently discovered that the pyranocoumarin compound decursin and its isomer decursinol angelate (DA) have potent novel antiandrogen receptor signaling activities. Because the ER and the androgen receptor belong to the steroid receptor superfamily, we examined whether these compounds affected ER expression and signaling in breast cancer cells.

Methods: We treated estrogen-dependent MCF-7 and estrogen-independent MDA MB-231 human breast cancer cells with decursin and DA, and examined cell growth, apoptosis, and ERalpha and ERbeta expression in both cell lines - and, in particular, estrogen-stimulated signaling in the MCF-7 cells. We compared these compounds with decursinol to determine their structure-activity relationship.

Results: Decursin and DA exerted growth inhibitory effects on MCF-7 cells through G1 arrest and caspase-mediated apoptosis. These compounds decreased ERalpha in MCF-7 cells at both mRNA and protein levels, and suppressed estrogen-stimulated genes. Decursin and the pure antiestrogen Faslodex exerted an additive growth inhibitory effect on MCF-7 cells. In MDA MB-231 cells, these compounds induced cell-cycle arrests in the G1 and G2 phases as well as inducing apoptosis, accompanied by an increased expression of ERbeta. In contrast, decursinol, which lacks the side chain of decursin and DA, did not have these cellular and molecular activities at comparable concentrations.

Conclusion: The side chain of decursin and DA is crucial for their anti-ER signaling and breast cancer growth inhibitory activities. These data provide mechanistic rationales for validating the chemopreventive and therapeutic efficacy of decursin and its derivatives in preclinical animal models of breast cancer.

Show MeSH
Related in: MedlinePlus