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Triptolide inhibits cell proliferation and tumorigenicity of human neuroblastoma cells.

Yan X, Ke XX, Zhao H, Huang M, Hu R, Cui H - Mol Med Rep (2014)

Bottom Line: Reverse transcription‑quantitative polymerase chain reaction was conducted to detect the expression levels of the apoptosis‑associated proteins, caspase‑3 and caspase‑9.The results demonstrated that exposure of BE(2)‑C human neuroblastoma cells to triptolide resulted in a reduction in cell growth and proliferation, and the induction of cell death and apoptosis, together with cell cycle arrest in the S phase.The xenograft experiment showed that triptolide significantly reduced tumor growth and development in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716, P.R. China.

ABSTRACT
Triptolide is a diterpene triepoxide, extracted from the Chinese herb Tripterygium wilfordii Hook F, which has been shown to have antitumor activity in a number of cancers. Neuroblastoma is an aggressive extracranial pediatric solid tumor, with significant chemotherapeutic resistance. In this study, triptolide was hypothesized to be a potential therapeutic agent for neuroblastoma. The effects of triptolide on neuroblastoma cell growth and tumor development were investigated. Cell growth and proliferation were evaluated using a cell counting kit‑8 assay and a 5-bromo-2-deoxyuridine staining assay. Cell cycle and apoptosis were detected by flow cytometry. Reverse transcription‑quantitative polymerase chain reaction was conducted to detect the expression levels of the apoptosis‑associated proteins, caspase‑3 and caspase‑9. The tumorigenicity of neuroblastoma cells was assessed by a soft agar clonogenic assay and an in vivo tumorigenic assay. The results demonstrated that exposure of BE(2)‑C human neuroblastoma cells to triptolide resulted in a reduction in cell growth and proliferation, and the induction of cell death and apoptosis, together with cell cycle arrest in the S phase. A soft agar assay indicated that triptolide inhibited the colony‑forming ability of BE(2)‑C neuroblastoma cells. The xenograft experiment showed that triptolide significantly reduced tumor growth and development in vivo. The data suggested that this Chinese herb may be a potential novel chemotherapeutic agent for neuroblastoma.

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Triptolide induced neuroblastoma cell apoptosis through caspase-9/caspase-3 pathway activation. Analysis of the percentage of (A) dead and (B) apoptotic BE(2)-C cells. BE(2)-C cells were treated with 25nM triptolide for 24 h, and cell death and apoptosis were determined by trypan blue dye and Annexin V-fluorescein isothiocyanate kit, respectively. DMSO was used as a control. (C) mRNA expression levels of caspase-3 and caspase-9 in BE(2)-C cells treated with DMSO or triptolide were determined by RT-qPCR analysis. Data represent the average obtained from three independent experiments. Data are presented as the mean ± standard deviation. *P<0.05 and **P<0.01, compared with control. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; DMSO, dimethyl sulfoxide.
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f4-mmr-11-02-0791: Triptolide induced neuroblastoma cell apoptosis through caspase-9/caspase-3 pathway activation. Analysis of the percentage of (A) dead and (B) apoptotic BE(2)-C cells. BE(2)-C cells were treated with 25nM triptolide for 24 h, and cell death and apoptosis were determined by trypan blue dye and Annexin V-fluorescein isothiocyanate kit, respectively. DMSO was used as a control. (C) mRNA expression levels of caspase-3 and caspase-9 in BE(2)-C cells treated with DMSO or triptolide were determined by RT-qPCR analysis. Data represent the average obtained from three independent experiments. Data are presented as the mean ± standard deviation. *P<0.05 and **P<0.01, compared with control. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; DMSO, dimethyl sulfoxide.

Mentions: It was also observed that exposure of BE(2)-C cells to triptolide could induce cell death and apoptosis. Triptolide significantly increased cell death from 1.88 in the control group to 25.9% in the triptolide 25 nM group (P<0.001; Fig. 4A). The apoptosis rate was also increased after treatment with 25 nM triptolide for 24 h (Fig. 4B). As shown in Fig. 4C and D, following triptolide treatment mRNA expression levels of caspase-9 and caspase-3 were increased 70.9 and 9.7 fold, respectively, compared with control. These results indicate that triptolide induces cell death and apoptosis through caspase-9 and caspase-3 activation.


Triptolide inhibits cell proliferation and tumorigenicity of human neuroblastoma cells.

Yan X, Ke XX, Zhao H, Huang M, Hu R, Cui H - Mol Med Rep (2014)

Triptolide induced neuroblastoma cell apoptosis through caspase-9/caspase-3 pathway activation. Analysis of the percentage of (A) dead and (B) apoptotic BE(2)-C cells. BE(2)-C cells were treated with 25nM triptolide for 24 h, and cell death and apoptosis were determined by trypan blue dye and Annexin V-fluorescein isothiocyanate kit, respectively. DMSO was used as a control. (C) mRNA expression levels of caspase-3 and caspase-9 in BE(2)-C cells treated with DMSO or triptolide were determined by RT-qPCR analysis. Data represent the average obtained from three independent experiments. Data are presented as the mean ± standard deviation. *P<0.05 and **P<0.01, compared with control. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; DMSO, dimethyl sulfoxide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262511&req=5

f4-mmr-11-02-0791: Triptolide induced neuroblastoma cell apoptosis through caspase-9/caspase-3 pathway activation. Analysis of the percentage of (A) dead and (B) apoptotic BE(2)-C cells. BE(2)-C cells were treated with 25nM triptolide for 24 h, and cell death and apoptosis were determined by trypan blue dye and Annexin V-fluorescein isothiocyanate kit, respectively. DMSO was used as a control. (C) mRNA expression levels of caspase-3 and caspase-9 in BE(2)-C cells treated with DMSO or triptolide were determined by RT-qPCR analysis. Data represent the average obtained from three independent experiments. Data are presented as the mean ± standard deviation. *P<0.05 and **P<0.01, compared with control. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; DMSO, dimethyl sulfoxide.
Mentions: It was also observed that exposure of BE(2)-C cells to triptolide could induce cell death and apoptosis. Triptolide significantly increased cell death from 1.88 in the control group to 25.9% in the triptolide 25 nM group (P<0.001; Fig. 4A). The apoptosis rate was also increased after treatment with 25 nM triptolide for 24 h (Fig. 4B). As shown in Fig. 4C and D, following triptolide treatment mRNA expression levels of caspase-9 and caspase-3 were increased 70.9 and 9.7 fold, respectively, compared with control. These results indicate that triptolide induces cell death and apoptosis through caspase-9 and caspase-3 activation.

Bottom Line: Reverse transcription‑quantitative polymerase chain reaction was conducted to detect the expression levels of the apoptosis‑associated proteins, caspase‑3 and caspase‑9.The results demonstrated that exposure of BE(2)‑C human neuroblastoma cells to triptolide resulted in a reduction in cell growth and proliferation, and the induction of cell death and apoptosis, together with cell cycle arrest in the S phase.The xenograft experiment showed that triptolide significantly reduced tumor growth and development in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716, P.R. China.

ABSTRACT
Triptolide is a diterpene triepoxide, extracted from the Chinese herb Tripterygium wilfordii Hook F, which has been shown to have antitumor activity in a number of cancers. Neuroblastoma is an aggressive extracranial pediatric solid tumor, with significant chemotherapeutic resistance. In this study, triptolide was hypothesized to be a potential therapeutic agent for neuroblastoma. The effects of triptolide on neuroblastoma cell growth and tumor development were investigated. Cell growth and proliferation were evaluated using a cell counting kit‑8 assay and a 5-bromo-2-deoxyuridine staining assay. Cell cycle and apoptosis were detected by flow cytometry. Reverse transcription‑quantitative polymerase chain reaction was conducted to detect the expression levels of the apoptosis‑associated proteins, caspase‑3 and caspase‑9. The tumorigenicity of neuroblastoma cells was assessed by a soft agar clonogenic assay and an in vivo tumorigenic assay. The results demonstrated that exposure of BE(2)‑C human neuroblastoma cells to triptolide resulted in a reduction in cell growth and proliferation, and the induction of cell death and apoptosis, together with cell cycle arrest in the S phase. A soft agar assay indicated that triptolide inhibited the colony‑forming ability of BE(2)‑C neuroblastoma cells. The xenograft experiment showed that triptolide significantly reduced tumor growth and development in vivo. The data suggested that this Chinese herb may be a potential novel chemotherapeutic agent for neuroblastoma.

Show MeSH
Related in: MedlinePlus