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Dual targeting of retinoid X receptor and histone deacetylase with DW22 as a novel antitumor approach.

Wang L, Chen G, Chen K, Ren Y, Li H, Jiang X, Jia L, Fu S, Li Y, Liu X, Wang S, Yang J, Wu C - Oncotarget (2015)

Bottom Line: We found that the co-expression of RXR-α and HDAC1 was frequently appeared in lung cancer and breast cancer tissues and cell lines.Moreover, intravenous administration of DW22 significantly retarded tumor growth of A549 and MDA-MB-435 xenograft mice models, and induced no substantial weight loss and gross toxicity.In addition, DW22 also reduced cell proliferation, angiogenesis, and induced cell apoptosis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, P.R. China.

ABSTRACT
Retinoid X receptor (RXR) and Histone deacetylase (HDAC) are considered important targets for cancer therapy due to their crucial roles in genetic or epigenetic regulations of cancer development and progression. Here, we evaluated the potential of dual targeting of RXR and HDAC using DW22 as a novel therapeutic approach to cancer treatment. We found that the co-expression of RXR-α and HDAC1 was frequently appeared in lung cancer and breast cancer tissues and cell lines. RXR was activated by DW22 in RXRα and HDAC1 overexpressed A549 and MDA-MB-435 cell lines. Meanwhile, DW22 inhibited the activity of HDAC by decreasing its expression in A549 and MDA-MB-435 cell lines, but not in RXRα and HDAC1 deficient cell lines. Moreover, DW22 suppressed cell growth, induced cell differentiation, prompted cell apoptosis and arrested cell cycle in A549, MDA-MB-435 or HL60 cell lines. Treatment human umbilical vascular endothelial cells (HUVECs) with DW22 suppressed migration, invasion and tube formation through decreasing VEGF expression. The up-regulation of Ac-H3 and p21, and down-regulation of VEGF caused by DW22 was markedly attenuated by silencing of HDAC1. Furthermore, knockdown of RXRα by siRNA completely blocked DW22-induced cell differentiation, but partially attenuated DW22-caused inhibition of cell proliferation, induction of cell apoptosis, and suppression of cell migration, invasion and tube formation. Moreover, intravenous administration of DW22 significantly retarded tumor growth of A549 and MDA-MB-435 xenograft mice models, and induced no substantial weight loss and gross toxicity. In addition, DW22 also reduced cell proliferation, angiogenesis, and induced cell apoptosis in vivo. Collectively, our data demonstrates that dual targeting of RXR and HDAC using DW22 possesses pleiotropic antitumor activities both in vitro and in vivo, providing a novel therapeutic approach for cancer treatment.

No MeSH data available.


Related in: MedlinePlus

The effects of DW22 on RXR activation and HDAC inhibition(A) 3D structure of Bexarotene, SAHA and DW22. (B)In vitro activation of RXR by DW22 in different concentrations (10 nM, 1 μM, and 50 μM). (C) Lanthascreen TR-FRET assay, demonstrating that DW22 increased the binding of the RXRα to coactivator peptide PGC1α in vitro. (D)In vitro inhibition of HDAC by DW22 in different concentrations (1 μM, 5 μM, and 20 μM). (E) Acetylated histone-3(Ac-H3, MW 17 kD), histone-3(H3, MW 17 kD), acetylated histone-4(Ac-H4, MW 10 kD), histone-4(H4, MW 11 kD), and HDAC1(MW 62 kD) were measured in A549 and MDA-MB-435 cell lines after DW22 (1, 5, 20 μM) treated for 48 h. β-actin expression was used as a loading control. All error bars are s.e.m. ***p < 0.005 compare with control group, *p < 0.05 compare with control group.
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Figure 2: The effects of DW22 on RXR activation and HDAC inhibition(A) 3D structure of Bexarotene, SAHA and DW22. (B)In vitro activation of RXR by DW22 in different concentrations (10 nM, 1 μM, and 50 μM). (C) Lanthascreen TR-FRET assay, demonstrating that DW22 increased the binding of the RXRα to coactivator peptide PGC1α in vitro. (D)In vitro inhibition of HDAC by DW22 in different concentrations (1 μM, 5 μM, and 20 μM). (E) Acetylated histone-3(Ac-H3, MW 17 kD), histone-3(H3, MW 17 kD), acetylated histone-4(Ac-H4, MW 10 kD), histone-4(H4, MW 11 kD), and HDAC1(MW 62 kD) were measured in A549 and MDA-MB-435 cell lines after DW22 (1, 5, 20 μM) treated for 48 h. β-actin expression was used as a loading control. All error bars are s.e.m. ***p < 0.005 compare with control group, *p < 0.05 compare with control group.

Mentions: DW22 was identified as a compound dual-targeting of RXR and HDAC [17] (See Figure 2A). Here, we examined the effect of DW22 on RXR activation using cell-based transactivation assays in RXR-α overexpressed cell lines A549 and MDA-MB-435. It was showed that treatment of A549 or MDA-MB-435 cells with DW22 significantly activated RXR reporter in a concentration-dependent manner (Figure 2B). As a positive control, Bexarotene (1 μM) treatment also resulted in an activation of RXR. To explore the activation mechanism, we detected the expression level of RXRα after treatment with DW22 in both cell lines. Western blot analysis data showed that either DW22 or Bexarotene had no effect on the expression of RXRα (Data not shown). These results demonstrate that DW22 can activate RXRα irrespective of its expression in A549 or MDA-MB-435 cells. The observations described above raise the possibility that DW22 might be an agonist of RXR. To test this hypothesis we examined the effect of DW22 on RXRα coactivator interaction in vitro by TR-FRET. In this assay, the interaction of the RXRα (indirectly labeled by terbium) with the coactivator peptide PGC1α (labeled with fluorescein) was detected. As shown in Figure 2C, DW22 treatment resulted in an enhanced binding of the RXRα to coactivator peptide PGC1α (EC50 = 3.6 nmol/L) compared to the well-studied RXR agonist, Bexarotene (EC50 = 16.2 nmol/L). These results suggest that DW22 is a ligand and an agonist of RXRα.


Dual targeting of retinoid X receptor and histone deacetylase with DW22 as a novel antitumor approach.

Wang L, Chen G, Chen K, Ren Y, Li H, Jiang X, Jia L, Fu S, Li Y, Liu X, Wang S, Yang J, Wu C - Oncotarget (2015)

The effects of DW22 on RXR activation and HDAC inhibition(A) 3D structure of Bexarotene, SAHA and DW22. (B)In vitro activation of RXR by DW22 in different concentrations (10 nM, 1 μM, and 50 μM). (C) Lanthascreen TR-FRET assay, demonstrating that DW22 increased the binding of the RXRα to coactivator peptide PGC1α in vitro. (D)In vitro inhibition of HDAC by DW22 in different concentrations (1 μM, 5 μM, and 20 μM). (E) Acetylated histone-3(Ac-H3, MW 17 kD), histone-3(H3, MW 17 kD), acetylated histone-4(Ac-H4, MW 10 kD), histone-4(H4, MW 11 kD), and HDAC1(MW 62 kD) were measured in A549 and MDA-MB-435 cell lines after DW22 (1, 5, 20 μM) treated for 48 h. β-actin expression was used as a loading control. All error bars are s.e.m. ***p < 0.005 compare with control group, *p < 0.05 compare with control group.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4496394&req=5

Figure 2: The effects of DW22 on RXR activation and HDAC inhibition(A) 3D structure of Bexarotene, SAHA and DW22. (B)In vitro activation of RXR by DW22 in different concentrations (10 nM, 1 μM, and 50 μM). (C) Lanthascreen TR-FRET assay, demonstrating that DW22 increased the binding of the RXRα to coactivator peptide PGC1α in vitro. (D)In vitro inhibition of HDAC by DW22 in different concentrations (1 μM, 5 μM, and 20 μM). (E) Acetylated histone-3(Ac-H3, MW 17 kD), histone-3(H3, MW 17 kD), acetylated histone-4(Ac-H4, MW 10 kD), histone-4(H4, MW 11 kD), and HDAC1(MW 62 kD) were measured in A549 and MDA-MB-435 cell lines after DW22 (1, 5, 20 μM) treated for 48 h. β-actin expression was used as a loading control. All error bars are s.e.m. ***p < 0.005 compare with control group, *p < 0.05 compare with control group.
Mentions: DW22 was identified as a compound dual-targeting of RXR and HDAC [17] (See Figure 2A). Here, we examined the effect of DW22 on RXR activation using cell-based transactivation assays in RXR-α overexpressed cell lines A549 and MDA-MB-435. It was showed that treatment of A549 or MDA-MB-435 cells with DW22 significantly activated RXR reporter in a concentration-dependent manner (Figure 2B). As a positive control, Bexarotene (1 μM) treatment also resulted in an activation of RXR. To explore the activation mechanism, we detected the expression level of RXRα after treatment with DW22 in both cell lines. Western blot analysis data showed that either DW22 or Bexarotene had no effect on the expression of RXRα (Data not shown). These results demonstrate that DW22 can activate RXRα irrespective of its expression in A549 or MDA-MB-435 cells. The observations described above raise the possibility that DW22 might be an agonist of RXR. To test this hypothesis we examined the effect of DW22 on RXRα coactivator interaction in vitro by TR-FRET. In this assay, the interaction of the RXRα (indirectly labeled by terbium) with the coactivator peptide PGC1α (labeled with fluorescein) was detected. As shown in Figure 2C, DW22 treatment resulted in an enhanced binding of the RXRα to coactivator peptide PGC1α (EC50 = 3.6 nmol/L) compared to the well-studied RXR agonist, Bexarotene (EC50 = 16.2 nmol/L). These results suggest that DW22 is a ligand and an agonist of RXRα.

Bottom Line: We found that the co-expression of RXR-α and HDAC1 was frequently appeared in lung cancer and breast cancer tissues and cell lines.Moreover, intravenous administration of DW22 significantly retarded tumor growth of A549 and MDA-MB-435 xenograft mice models, and induced no substantial weight loss and gross toxicity.In addition, DW22 also reduced cell proliferation, angiogenesis, and induced cell apoptosis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, P.R. China.

ABSTRACT
Retinoid X receptor (RXR) and Histone deacetylase (HDAC) are considered important targets for cancer therapy due to their crucial roles in genetic or epigenetic regulations of cancer development and progression. Here, we evaluated the potential of dual targeting of RXR and HDAC using DW22 as a novel therapeutic approach to cancer treatment. We found that the co-expression of RXR-α and HDAC1 was frequently appeared in lung cancer and breast cancer tissues and cell lines. RXR was activated by DW22 in RXRα and HDAC1 overexpressed A549 and MDA-MB-435 cell lines. Meanwhile, DW22 inhibited the activity of HDAC by decreasing its expression in A549 and MDA-MB-435 cell lines, but not in RXRα and HDAC1 deficient cell lines. Moreover, DW22 suppressed cell growth, induced cell differentiation, prompted cell apoptosis and arrested cell cycle in A549, MDA-MB-435 or HL60 cell lines. Treatment human umbilical vascular endothelial cells (HUVECs) with DW22 suppressed migration, invasion and tube formation through decreasing VEGF expression. The up-regulation of Ac-H3 and p21, and down-regulation of VEGF caused by DW22 was markedly attenuated by silencing of HDAC1. Furthermore, knockdown of RXRα by siRNA completely blocked DW22-induced cell differentiation, but partially attenuated DW22-caused inhibition of cell proliferation, induction of cell apoptosis, and suppression of cell migration, invasion and tube formation. Moreover, intravenous administration of DW22 significantly retarded tumor growth of A549 and MDA-MB-435 xenograft mice models, and induced no substantial weight loss and gross toxicity. In addition, DW22 also reduced cell proliferation, angiogenesis, and induced cell apoptosis in vivo. Collectively, our data demonstrates that dual targeting of RXR and HDAC using DW22 possesses pleiotropic antitumor activities both in vitro and in vivo, providing a novel therapeutic approach for cancer treatment.

No MeSH data available.


Related in: MedlinePlus