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Sulforaphane Increases Cyclin-Dependent Kinase Inhibitor, p21 Protein in Human Oral Carcinoma Cells and Nude Mouse Animal Model to Induce G(2)/M Cell Cycle Arrest.

Kim JH, Han Kwon K, Jung JY, Han HS, Hyun Shim J, Oh S, Choi KH, Choi ES, Shin JA, Leem DH, Soh Y, Cho NP, Cho SD - J Clin Biochem Nutr (2009)

Bottom Line: Previously, our group reported that sulforaphane (SFN), a naturally occurring chemopreventive agent from cruciferous vegetables, effectively inhibits the proliferation of KB and YD-10B human oral squamous carcinoma cells by causing apoptosis.Cell cycle arrest induced by SFN was associated with a significant increase in the p21 protein level and a decrease in cyclin B expression, but there was no change in the cyclin A protein level.In addition, SFN increased the p21 promoter activity significantly.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry and Institute of Oral Bioscience, Brain Korea 21 project, Chonbuk National University, Jeonju, 561-756, Republic of Korea.

ABSTRACT
Previously, our group reported that sulforaphane (SFN), a naturally occurring chemopreventive agent from cruciferous vegetables, effectively inhibits the proliferation of KB and YD-10B human oral squamous carcinoma cells by causing apoptosis. In this study, treatment of 20 and 40 microM of SFN for 12 h caused a cell cycle arrest in the G(2)/M phase. Cell cycle arrest induced by SFN was associated with a significant increase in the p21 protein level and a decrease in cyclin B expression, but there was no change in the cyclin A protein level. In addition, SFN increased the p21 promoter activity significantly. Furthermore, SFN induced p21 protein expression in a nude mouse xenograft model suggesting that SFN is a potent inducer of the p21 protein in human oral squamous carcinoma cells. These findings show that SFN is a promising candidate for molecular-targeting chemotherapy against human oral squamous cell carcinoma.

No MeSH data available.


Related in: MedlinePlus

Effects of SFN on the levels of the proteins involved in the regulation of G2/M transition. (A) immunoblotting for p21, cyclin B and cyclin A using lysates from control and SFN-treated KB cells. (B) The data points in the graph are the mean ± SD of three independent experiments. *p<0.05 compared to the control group. (C) Transactivation activity of pWWP (p21 promoter construct) induced by SFN in KB cells. (D) immunobloting for sp1 protein using lysates from control and SFN-treated KB cells.
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Figure 3: Effects of SFN on the levels of the proteins involved in the regulation of G2/M transition. (A) immunoblotting for p21, cyclin B and cyclin A using lysates from control and SFN-treated KB cells. (B) The data points in the graph are the mean ± SD of three independent experiments. *p<0.05 compared to the control group. (C) Transactivation activity of pWWP (p21 promoter construct) induced by SFN in KB cells. (D) immunobloting for sp1 protein using lysates from control and SFN-treated KB cells.

Mentions: Because, the most of the cells were arrested at the G2/M phase, experiments were carried out to determine whether the SFN treatment also affects the expression level of G2/M phase cell cycle regulatory proteins. KB cells were treated with 20 µM of SFN for 12 h and the expression of these proteins (p21, cyclin A and cyclin B) were examined by western blot analysis. Fig. 3A and B showed a significant increase in the level of p21 protein expression and the level of cyclin B expression was significantly reduced by SFN whereas cyclin A was not changed. Furthermore, SFN induced the transactivation in KB cells transfected with pWWP (Fig. 3C). To find whether sp1 protein regulates p21 protein expression, we evaluated the level of sp1 protein expression. Unfortunately, sp1 protein was not changed by SFN compared to control (Fig. 3D). These results suggest that SFN modulates the expression level of the G2/M phase cell cycle regulatory proteins and its transactivation in KB oral carcinoma cells.


Sulforaphane Increases Cyclin-Dependent Kinase Inhibitor, p21 Protein in Human Oral Carcinoma Cells and Nude Mouse Animal Model to Induce G(2)/M Cell Cycle Arrest.

Kim JH, Han Kwon K, Jung JY, Han HS, Hyun Shim J, Oh S, Choi KH, Choi ES, Shin JA, Leem DH, Soh Y, Cho NP, Cho SD - J Clin Biochem Nutr (2009)

Effects of SFN on the levels of the proteins involved in the regulation of G2/M transition. (A) immunoblotting for p21, cyclin B and cyclin A using lysates from control and SFN-treated KB cells. (B) The data points in the graph are the mean ± SD of three independent experiments. *p<0.05 compared to the control group. (C) Transactivation activity of pWWP (p21 promoter construct) induced by SFN in KB cells. (D) immunobloting for sp1 protein using lysates from control and SFN-treated KB cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: Effects of SFN on the levels of the proteins involved in the regulation of G2/M transition. (A) immunoblotting for p21, cyclin B and cyclin A using lysates from control and SFN-treated KB cells. (B) The data points in the graph are the mean ± SD of three independent experiments. *p<0.05 compared to the control group. (C) Transactivation activity of pWWP (p21 promoter construct) induced by SFN in KB cells. (D) immunobloting for sp1 protein using lysates from control and SFN-treated KB cells.
Mentions: Because, the most of the cells were arrested at the G2/M phase, experiments were carried out to determine whether the SFN treatment also affects the expression level of G2/M phase cell cycle regulatory proteins. KB cells were treated with 20 µM of SFN for 12 h and the expression of these proteins (p21, cyclin A and cyclin B) were examined by western blot analysis. Fig. 3A and B showed a significant increase in the level of p21 protein expression and the level of cyclin B expression was significantly reduced by SFN whereas cyclin A was not changed. Furthermore, SFN induced the transactivation in KB cells transfected with pWWP (Fig. 3C). To find whether sp1 protein regulates p21 protein expression, we evaluated the level of sp1 protein expression. Unfortunately, sp1 protein was not changed by SFN compared to control (Fig. 3D). These results suggest that SFN modulates the expression level of the G2/M phase cell cycle regulatory proteins and its transactivation in KB oral carcinoma cells.

Bottom Line: Previously, our group reported that sulforaphane (SFN), a naturally occurring chemopreventive agent from cruciferous vegetables, effectively inhibits the proliferation of KB and YD-10B human oral squamous carcinoma cells by causing apoptosis.Cell cycle arrest induced by SFN was associated with a significant increase in the p21 protein level and a decrease in cyclin B expression, but there was no change in the cyclin A protein level.In addition, SFN increased the p21 promoter activity significantly.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry and Institute of Oral Bioscience, Brain Korea 21 project, Chonbuk National University, Jeonju, 561-756, Republic of Korea.

ABSTRACT
Previously, our group reported that sulforaphane (SFN), a naturally occurring chemopreventive agent from cruciferous vegetables, effectively inhibits the proliferation of KB and YD-10B human oral squamous carcinoma cells by causing apoptosis. In this study, treatment of 20 and 40 microM of SFN for 12 h caused a cell cycle arrest in the G(2)/M phase. Cell cycle arrest induced by SFN was associated with a significant increase in the p21 protein level and a decrease in cyclin B expression, but there was no change in the cyclin A protein level. In addition, SFN increased the p21 promoter activity significantly. Furthermore, SFN induced p21 protein expression in a nude mouse xenograft model suggesting that SFN is a potent inducer of the p21 protein in human oral squamous carcinoma cells. These findings show that SFN is a promising candidate for molecular-targeting chemotherapy against human oral squamous cell carcinoma.

No MeSH data available.


Related in: MedlinePlus