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Effect of celastrol on growth inhibition of prostate cancer cells through the regulation of hERG channel in vitro.

Ji N, Li J, Wei Z, Kong F, Jin H, Chen X, Li Y, Deng Y - Biomed Res Int (2015)

Bottom Line: Moreover, Celastrol induced DU145 cell apoptosis in a cell cycle-dependent manner, which means Celastrol could arrest DU145 cells in G0/G1 phase; accordingly, cells in S phase decreased gradually and no obvious changes were found in G2/M phase cells.Through transmission electron microscope, apoptotic bodies containing nuclear fragments were found in Celastrol-treated DU145 cells.Overexpression of hERG channel was found in DU145 cells, while Celastrol could downregulate it at both protein and mRNA level in a dose-dependent manner (P < 0.01).

View Article: PubMed Central - PubMed

Affiliation: Medical College, Wuhan University of Science and Technology, Wuhan 430065, China ; The Outpatient Department, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China.

ABSTRACT

Objective: To explore the antiprostate cancer effects of Celastrol on prostate cancer cells' proliferation, apoptosis, and cell cycle distribution, as well as the correlation to the regulation of hERG.

Methods: DU145 cells were treated with various concentrations of Celastrol (0.25-16.0 μmol/L) for 0-72 hours. MTT assay was used to evaluate the inhibition effect of Celastrol on the growth of DU145 cells. Cell apoptosis was detected through both Annexin-V FITC/PI double-labeled cytometry and Hoechst 33258. Cell cycle regulation was examined by a propidium iodide method. Western blot and RT-PCR technologies were applied to assess the expression level of hERG in DU145 cells.

Results: Celastrol presented striking growth inhibition and apoptosis induction potency on DU145 cells in vitro in a time- and dose-dependent manner. The IC50 value of Celastrol for 24 hours was 2.349 ± 0.213 μmol/L. Moreover, Celastrol induced DU145 cell apoptosis in a cell cycle-dependent manner, which means Celastrol could arrest DU145 cells in G0/G1 phase; accordingly, cells in S phase decreased gradually and no obvious changes were found in G2/M phase cells. Through transmission electron microscope, apoptotic bodies containing nuclear fragments were found in Celastrol-treated DU145 cells. Overexpression of hERG channel was found in DU145 cells, while Celastrol could downregulate it at both protein and mRNA level in a dose-dependent manner (P < 0.01).

Conclusions: Celastrol exhibits its antiprostate cancer effects partially through the downregulation of the expression level of hERG channel in DU145 cells, suggesting that Celastrol may be a potential agent against prostate cancer with a mechanism of blocking the hERG channel.

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(a) Effects of Celastrol on the expression of hERG protein in DU145 cells and normal mononuclear cells with various concentrations for 24 h. (b) Effects of Celastrol on the expression of hERG mRNA in DU145 cells and normal mononuclear cells with various concentrations for 24 h.
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fig4: (a) Effects of Celastrol on the expression of hERG protein in DU145 cells and normal mononuclear cells with various concentrations for 24 h. (b) Effects of Celastrol on the expression of hERG mRNA in DU145 cells and normal mononuclear cells with various concentrations for 24 h.

Mentions: Compared with normal mononuclear cells, the presence of hERG potassium channel protein expression levels was higher in DU145 cells and the 0.5~2.0 μmol/L tripterine was treated for 24 h. The protein expression had a concentration dependent decline, with a statistically significant difference (P < 0.05). In order to further clarify the role of Tripterygium wilfordii red on the hERG protein, we examined the changes of hERG protein and mRNA content in the level of gene transcription. Similarly, hERG potassium channel protein level of mRNA was dose-dependently downregulated and obviously higher than the mononuclear cells of normal hERG gene expression level (Figure 4).


Effect of celastrol on growth inhibition of prostate cancer cells through the regulation of hERG channel in vitro.

Ji N, Li J, Wei Z, Kong F, Jin H, Chen X, Li Y, Deng Y - Biomed Res Int (2015)

(a) Effects of Celastrol on the expression of hERG protein in DU145 cells and normal mononuclear cells with various concentrations for 24 h. (b) Effects of Celastrol on the expression of hERG mRNA in DU145 cells and normal mononuclear cells with various concentrations for 24 h.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383143&req=5

fig4: (a) Effects of Celastrol on the expression of hERG protein in DU145 cells and normal mononuclear cells with various concentrations for 24 h. (b) Effects of Celastrol on the expression of hERG mRNA in DU145 cells and normal mononuclear cells with various concentrations for 24 h.
Mentions: Compared with normal mononuclear cells, the presence of hERG potassium channel protein expression levels was higher in DU145 cells and the 0.5~2.0 μmol/L tripterine was treated for 24 h. The protein expression had a concentration dependent decline, with a statistically significant difference (P < 0.05). In order to further clarify the role of Tripterygium wilfordii red on the hERG protein, we examined the changes of hERG protein and mRNA content in the level of gene transcription. Similarly, hERG potassium channel protein level of mRNA was dose-dependently downregulated and obviously higher than the mononuclear cells of normal hERG gene expression level (Figure 4).

Bottom Line: Moreover, Celastrol induced DU145 cell apoptosis in a cell cycle-dependent manner, which means Celastrol could arrest DU145 cells in G0/G1 phase; accordingly, cells in S phase decreased gradually and no obvious changes were found in G2/M phase cells.Through transmission electron microscope, apoptotic bodies containing nuclear fragments were found in Celastrol-treated DU145 cells.Overexpression of hERG channel was found in DU145 cells, while Celastrol could downregulate it at both protein and mRNA level in a dose-dependent manner (P < 0.01).

View Article: PubMed Central - PubMed

Affiliation: Medical College, Wuhan University of Science and Technology, Wuhan 430065, China ; The Outpatient Department, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China.

ABSTRACT

Objective: To explore the antiprostate cancer effects of Celastrol on prostate cancer cells' proliferation, apoptosis, and cell cycle distribution, as well as the correlation to the regulation of hERG.

Methods: DU145 cells were treated with various concentrations of Celastrol (0.25-16.0 μmol/L) for 0-72 hours. MTT assay was used to evaluate the inhibition effect of Celastrol on the growth of DU145 cells. Cell apoptosis was detected through both Annexin-V FITC/PI double-labeled cytometry and Hoechst 33258. Cell cycle regulation was examined by a propidium iodide method. Western blot and RT-PCR technologies were applied to assess the expression level of hERG in DU145 cells.

Results: Celastrol presented striking growth inhibition and apoptosis induction potency on DU145 cells in vitro in a time- and dose-dependent manner. The IC50 value of Celastrol for 24 hours was 2.349 ± 0.213 μmol/L. Moreover, Celastrol induced DU145 cell apoptosis in a cell cycle-dependent manner, which means Celastrol could arrest DU145 cells in G0/G1 phase; accordingly, cells in S phase decreased gradually and no obvious changes were found in G2/M phase cells. Through transmission electron microscope, apoptotic bodies containing nuclear fragments were found in Celastrol-treated DU145 cells. Overexpression of hERG channel was found in DU145 cells, while Celastrol could downregulate it at both protein and mRNA level in a dose-dependent manner (P < 0.01).

Conclusions: Celastrol exhibits its antiprostate cancer effects partially through the downregulation of the expression level of hERG channel in DU145 cells, suggesting that Celastrol may be a potential agent against prostate cancer with a mechanism of blocking the hERG channel.

Show MeSH
Related in: MedlinePlus