Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes: Association with c-myc, p53, and bcl-2 mRNA

M. J. Hsu; J. S. Cheng; H. C. Huang

doi:10.1038/sj.bjp.0703559

Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes: Association with c-myc, p53, and bcl-2 mRNA

M. J. Hsu, J. S. Cheng, H. C. Huang

Research output: Contribution to journal › Article › peer-review

53 Citations (Scopus)

Abstract

1. The mechanisms involved in the apoptotic effect of saikosaponin-d, a triterpene saponin from Bupleurum falcatum L., were studied in human CEM lymphocytes and compared with those of dexamethasone (3 / 10^-7 M). 2. Saikosaponin-d (10^-8 to 10^-5 M) inhibited the serum-stimulated [³H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [³H]-thymidine incorporation. 3. Cell viability was unaffected by saikosaponin-d until 10^-5-10^-4 M. Dexamethasone significantly reduced the number of viable cells. 4. Following saikosaponin-d (10^-5-10^-4 M) treatment, flow cytometry analysis of propidium iodide-stained cells showed a significant increase in the percentage of cells in the apoptotic region. Dexamethasone also significantly increased the percentage of apoptotic cells. The supravital exposure to propidium iodide and annexin V labelling demonstrated that saikosaponin-d (10^-5-10^-4 M) induced apoptosis as well as necrosis. 5. The apoptotic effect of saikosaponin-d (3 / 10^-6-10^-4 M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 / 10^-6-10^-5 M) was unaffected by the presence of Z-VAD-FMK, indicating that saikosaponin-d-induced apoptosis may not be mediated by caspase activity. However, the percentage of apoptotic cells induced by dexamethasone was significantly reduced by the presence of Z-VAD-FMK. 6. Levels of c-myc, p53, and bcl-2 mRNA were analysed by the reverse transcription-polymerase chain reaction. Levels of c-myc and p53 mRNA were significantly increased, while the level of bcl-2 mRNA was decreased, by saikosaponin-d (10^-5 M) treatment. Dexamethasone did not significantly change the expression of these genes. 7. It is suggested that the apoptotic effect of saikosaponin-d may be partly mediated by increases in c-myc and p53 mRNA levels accompanied by a decrease in bcl-2 mRNA level.

Original language	English
Pages (from-to)	1285-1293
Number of pages	9
Journal	British Journal of Pharmacology
Volume	131
Issue number	7
DOIs	https://doi.org/10.1038/sj.bjp.0703559
Publication status	Published - 2000

Keywords

Apoptosis
CEM lymphocytes
Caspase
Saikosaponin-d
bcl-2
c-myc
p53

ASJC Scopus subject areas

Pharmacology

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10.1038/sj.bjp.0703559

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title = "Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes: Association with c-myc, p53, and bcl-2 mRNA",

abstract = "1. The mechanisms involved in the apoptotic effect of saikosaponin-d, a triterpene saponin from Bupleurum falcatum L., were studied in human CEM lymphocytes and compared with those of dexamethasone (3 / 10-7 M). 2. Saikosaponin-d (10-8 to 10-5 M) inhibited the serum-stimulated [3H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [3H]-thymidine incorporation. 3. Cell viability was unaffected by saikosaponin-d until 10-5-10-4 M. Dexamethasone significantly reduced the number of viable cells. 4. Following saikosaponin-d (10-5-10-4 M) treatment, flow cytometry analysis of propidium iodide-stained cells showed a significant increase in the percentage of cells in the apoptotic region. Dexamethasone also significantly increased the percentage of apoptotic cells. The supravital exposure to propidium iodide and annexin V labelling demonstrated that saikosaponin-d (10-5-10-4 M) induced apoptosis as well as necrosis. 5. The apoptotic effect of saikosaponin-d (3 / 10-6-10-4 M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 / 10-6-10-5 M) was unaffected by the presence of Z-VAD-FMK, indicating that saikosaponin-d-induced apoptosis may not be mediated by caspase activity. However, the percentage of apoptotic cells induced by dexamethasone was significantly reduced by the presence of Z-VAD-FMK. 6. Levels of c-myc, p53, and bcl-2 mRNA were analysed by the reverse transcription-polymerase chain reaction. Levels of c-myc and p53 mRNA were significantly increased, while the level of bcl-2 mRNA was decreased, by saikosaponin-d (10-5 M) treatment. Dexamethasone did not significantly change the expression of these genes. 7. It is suggested that the apoptotic effect of saikosaponin-d may be partly mediated by increases in c-myc and p53 mRNA levels accompanied by a decrease in bcl-2 mRNA level.",

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author = "Hsu, {M. J.} and Cheng, {J. S.} and Huang, {H. C.}",

year = "2000",

doi = "10.1038/sj.bjp.0703559",

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AU - Cheng, J. S.

AU - Huang, H. C.

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N2 - 1. The mechanisms involved in the apoptotic effect of saikosaponin-d, a triterpene saponin from Bupleurum falcatum L., were studied in human CEM lymphocytes and compared with those of dexamethasone (3 / 10-7 M). 2. Saikosaponin-d (10-8 to 10-5 M) inhibited the serum-stimulated [3H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [3H]-thymidine incorporation. 3. Cell viability was unaffected by saikosaponin-d until 10-5-10-4 M. Dexamethasone significantly reduced the number of viable cells. 4. Following saikosaponin-d (10-5-10-4 M) treatment, flow cytometry analysis of propidium iodide-stained cells showed a significant increase in the percentage of cells in the apoptotic region. Dexamethasone also significantly increased the percentage of apoptotic cells. The supravital exposure to propidium iodide and annexin V labelling demonstrated that saikosaponin-d (10-5-10-4 M) induced apoptosis as well as necrosis. 5. The apoptotic effect of saikosaponin-d (3 / 10-6-10-4 M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 / 10-6-10-5 M) was unaffected by the presence of Z-VAD-FMK, indicating that saikosaponin-d-induced apoptosis may not be mediated by caspase activity. However, the percentage of apoptotic cells induced by dexamethasone was significantly reduced by the presence of Z-VAD-FMK. 6. Levels of c-myc, p53, and bcl-2 mRNA were analysed by the reverse transcription-polymerase chain reaction. Levels of c-myc and p53 mRNA were significantly increased, while the level of bcl-2 mRNA was decreased, by saikosaponin-d (10-5 M) treatment. Dexamethasone did not significantly change the expression of these genes. 7. It is suggested that the apoptotic effect of saikosaponin-d may be partly mediated by increases in c-myc and p53 mRNA levels accompanied by a decrease in bcl-2 mRNA level.

AB - 1. The mechanisms involved in the apoptotic effect of saikosaponin-d, a triterpene saponin from Bupleurum falcatum L., were studied in human CEM lymphocytes and compared with those of dexamethasone (3 / 10-7 M). 2. Saikosaponin-d (10-8 to 10-5 M) inhibited the serum-stimulated [3H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [3H]-thymidine incorporation. 3. Cell viability was unaffected by saikosaponin-d until 10-5-10-4 M. Dexamethasone significantly reduced the number of viable cells. 4. Following saikosaponin-d (10-5-10-4 M) treatment, flow cytometry analysis of propidium iodide-stained cells showed a significant increase in the percentage of cells in the apoptotic region. Dexamethasone also significantly increased the percentage of apoptotic cells. The supravital exposure to propidium iodide and annexin V labelling demonstrated that saikosaponin-d (10-5-10-4 M) induced apoptosis as well as necrosis. 5. The apoptotic effect of saikosaponin-d (3 / 10-6-10-4 M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 / 10-6-10-5 M) was unaffected by the presence of Z-VAD-FMK, indicating that saikosaponin-d-induced apoptosis may not be mediated by caspase activity. However, the percentage of apoptotic cells induced by dexamethasone was significantly reduced by the presence of Z-VAD-FMK. 6. Levels of c-myc, p53, and bcl-2 mRNA were analysed by the reverse transcription-polymerase chain reaction. Levels of c-myc and p53 mRNA were significantly increased, while the level of bcl-2 mRNA was decreased, by saikosaponin-d (10-5 M) treatment. Dexamethasone did not significantly change the expression of these genes. 7. It is suggested that the apoptotic effect of saikosaponin-d may be partly mediated by increases in c-myc and p53 mRNA levels accompanied by a decrease in bcl-2 mRNA level.

KW - Apoptosis

KW - CEM lymphocytes

KW - Caspase

KW - Saikosaponin-d

KW - bcl-2

KW - c-myc

KW - p53

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Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes: Association with c-myc, p53, and bcl-2 mRNA

Abstract

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