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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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BrdU immunofluorescence staining assay. (A) Immunofluorescence staining of Brdu in BE(2)-C cells treated with triptolide (25 and 50 nM) for 24 h. Scale bars, 25 μm. (B) The percentage of BrdU positive cells from panel A was calculated. Each value represents 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. BrdU, 5-bromo-2-deoxyuridine; DMSO, dimethyl sulfoxide.
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f2-mmr-11-02-0791: BrdU immunofluorescence staining assay. (A) Immunofluorescence staining of Brdu in BE(2)-C cells treated with triptolide (25 and 50 nM) for 24 h. Scale bars, 25 μm. (B) The percentage of BrdU positive cells from panel A was calculated. Each value represents 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. BrdU, 5-bromo-2-deoxyuridine; DMSO, dimethyl sulfoxide.

Mentions: BE(2)-C cells were treated with increasing doses of triptolide for 24 h. A concentration-dependent response to triptolide in the BE(2)-C cells was observed. As shown in Fig. 1A, triptolide inhibited cell growth even at a low dose of 5 nM. The cell viability was significantly reduced to 50% at 50 nM of triptolide. Triptolide also inhibited cell growth in a time dependent manner (Fig. 1B and C). Moreover, immunofluorescent staining using a BrdU label confirmed that triptolide markedly inhibited cell proliferation (Fig. 2A and B).


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)

BrdU immunofluorescence staining assay. (A) Immunofluorescence staining of Brdu in BE(2)-C cells treated with triptolide (25 and 50 nM) for 24 h. Scale bars, 25 μm. (B) The percentage of BrdU positive cells from panel A was calculated. Each value represents 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. BrdU, 5-bromo-2-deoxyuridine; DMSO, dimethyl sulfoxide.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2-mmr-11-02-0791: BrdU immunofluorescence staining assay. (A) Immunofluorescence staining of Brdu in BE(2)-C cells treated with triptolide (25 and 50 nM) for 24 h. Scale bars, 25 μm. (B) The percentage of BrdU positive cells from panel A was calculated. Each value represents 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. BrdU, 5-bromo-2-deoxyuridine; DMSO, dimethyl sulfoxide.
Mentions: BE(2)-C cells were treated with increasing doses of triptolide for 24 h. A concentration-dependent response to triptolide in the BE(2)-C cells was observed. As shown in Fig. 1A, triptolide inhibited cell growth even at a low dose of 5 nM. The cell viability was significantly reduced to 50% at 50 nM of triptolide. Triptolide also inhibited cell growth in a time dependent manner (Fig. 1B and C). Moreover, immunofluorescent staining using a BrdU label confirmed that triptolide markedly inhibited cell proliferation (Fig. 2A and B).

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