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Platycodin D induces apoptosis and triggers ERK- and JNK-mediated autophagy in human hepatocellular carcinoma BEL-7402 cells.

Li T, Xu XH, Tang ZH, Wang YF, Leung CH, Ma DL, Chen XP, Wang YT, Chen Y, Lu JJ - Acta Pharmacol. Sin. (2015)

Bottom Line: Moreover, platycodin D (20 μmol/L) activated the ERK and JNK pathways in BEL-7402 cells, and simultaneous blockage of the two pathways effectively suppressed platycodin D-induced autophagy and enhanced platycodin D-induced apoptosis.In BEL-7402-bearing mice, platycodin D (10 mg·kg(-1)•d(-1)) significantly reduced relative tumor volume with decreased body weight.Platycodin D-induced cell proliferation inhibition and apoptosis are amplified by co-treatment with autophagy inhibitors.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.

ABSTRACT

Aim: Platycodin D, the main saponin isolated from Chinese herb Platycodonis Radix, exhibits anticancer activities against various cancer cell lines. Here we evaluated its anticancer action against human hepatocellular carcinoma cells in vitro and in vivo, and elucidated the relationship between platycodin D-induced apoptosis and autophagy.

Methods: The viability of human hepatocellular carcinoma BEL-7402 cells was evaluated with MTT assay, and the apoptosis was examined using Annexin V/PI and Hoechst 33342 staining assays. Monodansylcadaverine (MDC) staining was used to label autophagic vacuoles. The proteins were detected using Western blot analysis. For studying its anticancer action in vivo, platycodin D (5 and 10 mg· kg(-1)·d(-1)) was intraperitoneally injected to BEL-7402-bearing mice for 21 days.

Results: Platycodin D (5-40 μmol/L) inhibited the cell proliferation in vitro with IC50 values of 37.70±3.99, 24.30±2.30 and 19.70±2.36 μmol/L at 24, 48 and 72 h, respectively. Platycodin D (5-20 μmol/L) dose-dependently increased BEL-7402 cell apoptosis, increased the Bax/Bcl-2 ratio and the levels of cleaved PARP and cleaved caspase-3, and decreased the level of Bcl-2. Furthermore, platycodin D (5-20 μmol/L) induced autophagy in BEL-7402 cells, as evidenced by formation of cytoplasmic vacuoles, increased amounts of LC3-II, and increased numbers of MDC-positive cells. Pretreatment with the autophagy inhibitor chloroquine (5 μmol/L) or BAF (50 nmol/L) significantly enhanced platycodin D-induced proliferation inhibition and apoptosis. Moreover, platycodin D (20 μmol/L) activated the ERK and JNK pathways in BEL-7402 cells, and simultaneous blockage of the two pathways effectively suppressed platycodin D-induced autophagy and enhanced platycodin D-induced apoptosis. In BEL-7402-bearing mice, platycodin D (10 mg·kg(-1)•d(-1)) significantly reduced relative tumor volume with decreased body weight.

Conclusion: Platycodin D not only inhibits the proliferation of BEL-7402 cells but also suppresses BEL-7402 xenograft tumor growth. Platycodin D-induced cell proliferation inhibition and apoptosis are amplified by co-treatment with autophagy inhibitors.

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PD inhibits the proliferation of hepatocellular carcinoma BEL-7402 cells. (A) The chemical structure of PD. (B) Cells were treated with different concentrations of PD for 24, 48, and 72 h, and cell proliferation inhibition was detected by the MTT assay. Statistical significance was analyzed using one-way analysis of variance using Graph Pad Prism (Demo, Version 5) with bP<0.05, cP<0.01 vs control.
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fig1: PD inhibits the proliferation of hepatocellular carcinoma BEL-7402 cells. (A) The chemical structure of PD. (B) Cells were treated with different concentrations of PD for 24, 48, and 72 h, and cell proliferation inhibition was detected by the MTT assay. Statistical significance was analyzed using one-way analysis of variance using Graph Pad Prism (Demo, Version 5) with bP<0.05, cP<0.01 vs control.

Mentions: Platycodon grandiflorum (Jacq.) A. DC, commonly known as the balloon flower, is widely distributed in Northeast Asia. Platycodonis radix is the two- or three-year-old root of Platycodon grandiflorum (Jacq.) A. DC, with a long history of use as a dietary source and a folk remedy for pulmonary diseases and respiratory system disorders in Korea, Japan and China1. Platycodin D (PD) (Figure 1A) is one of the main saponins extracted from Platycodonis radix, and it possesses immune-stimulatory2, anti-inflammatory3,4, anti-nociceptive5, anti-obesity5,6, and anti-atherogenic7 activities. In particular, PD exhibits excellent anticancer effects against various cancer cell lines mainly by inhibiting cell proliferation, inducing cell cycle arrest and promoting apoptosis8,9,10,11,12,13,14. PD-induced G2/M phase cycle arrest may be regulated by suppressing spindle microtubule dynamics in leukemia U937, THP-1, and K562 cells11. PD-mediated apoptosis may be related to the activation of caspase 3 and the induction of reactive oxygen species12. In our previous studies, PD inhibited cell proliferation and induced apoptosis via the induction of poly ADP-ribose polymerase (PARP) cleavage, the up-regulation of Bax and the down-regulation of survivin in hepatocellular carcinoma cells15. In addition, PD also triggered autophagy in a broad spectrum of cell lines including breast cancer, lung cancer, and hepatocellular carcinoma cells16.


Platycodin D induces apoptosis and triggers ERK- and JNK-mediated autophagy in human hepatocellular carcinoma BEL-7402 cells.

Li T, Xu XH, Tang ZH, Wang YF, Leung CH, Ma DL, Chen XP, Wang YT, Chen Y, Lu JJ - Acta Pharmacol. Sin. (2015)

PD inhibits the proliferation of hepatocellular carcinoma BEL-7402 cells. (A) The chemical structure of PD. (B) Cells were treated with different concentrations of PD for 24, 48, and 72 h, and cell proliferation inhibition was detected by the MTT assay. Statistical significance was analyzed using one-way analysis of variance using Graph Pad Prism (Demo, Version 5) with bP<0.05, cP<0.01 vs control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4816242&req=5

fig1: PD inhibits the proliferation of hepatocellular carcinoma BEL-7402 cells. (A) The chemical structure of PD. (B) Cells were treated with different concentrations of PD for 24, 48, and 72 h, and cell proliferation inhibition was detected by the MTT assay. Statistical significance was analyzed using one-way analysis of variance using Graph Pad Prism (Demo, Version 5) with bP<0.05, cP<0.01 vs control.
Mentions: Platycodon grandiflorum (Jacq.) A. DC, commonly known as the balloon flower, is widely distributed in Northeast Asia. Platycodonis radix is the two- or three-year-old root of Platycodon grandiflorum (Jacq.) A. DC, with a long history of use as a dietary source and a folk remedy for pulmonary diseases and respiratory system disorders in Korea, Japan and China1. Platycodin D (PD) (Figure 1A) is one of the main saponins extracted from Platycodonis radix, and it possesses immune-stimulatory2, anti-inflammatory3,4, anti-nociceptive5, anti-obesity5,6, and anti-atherogenic7 activities. In particular, PD exhibits excellent anticancer effects against various cancer cell lines mainly by inhibiting cell proliferation, inducing cell cycle arrest and promoting apoptosis8,9,10,11,12,13,14. PD-induced G2/M phase cycle arrest may be regulated by suppressing spindle microtubule dynamics in leukemia U937, THP-1, and K562 cells11. PD-mediated apoptosis may be related to the activation of caspase 3 and the induction of reactive oxygen species12. In our previous studies, PD inhibited cell proliferation and induced apoptosis via the induction of poly ADP-ribose polymerase (PARP) cleavage, the up-regulation of Bax and the down-regulation of survivin in hepatocellular carcinoma cells15. In addition, PD also triggered autophagy in a broad spectrum of cell lines including breast cancer, lung cancer, and hepatocellular carcinoma cells16.

Bottom Line: Moreover, platycodin D (20 μmol/L) activated the ERK and JNK pathways in BEL-7402 cells, and simultaneous blockage of the two pathways effectively suppressed platycodin D-induced autophagy and enhanced platycodin D-induced apoptosis.In BEL-7402-bearing mice, platycodin D (10 mg·kg(-1)•d(-1)) significantly reduced relative tumor volume with decreased body weight.Platycodin D-induced cell proliferation inhibition and apoptosis are amplified by co-treatment with autophagy inhibitors.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.

ABSTRACT

Aim: Platycodin D, the main saponin isolated from Chinese herb Platycodonis Radix, exhibits anticancer activities against various cancer cell lines. Here we evaluated its anticancer action against human hepatocellular carcinoma cells in vitro and in vivo, and elucidated the relationship between platycodin D-induced apoptosis and autophagy.

Methods: The viability of human hepatocellular carcinoma BEL-7402 cells was evaluated with MTT assay, and the apoptosis was examined using Annexin V/PI and Hoechst 33342 staining assays. Monodansylcadaverine (MDC) staining was used to label autophagic vacuoles. The proteins were detected using Western blot analysis. For studying its anticancer action in vivo, platycodin D (5 and 10 mg· kg(-1)·d(-1)) was intraperitoneally injected to BEL-7402-bearing mice for 21 days.

Results: Platycodin D (5-40 μmol/L) inhibited the cell proliferation in vitro with IC50 values of 37.70±3.99, 24.30±2.30 and 19.70±2.36 μmol/L at 24, 48 and 72 h, respectively. Platycodin D (5-20 μmol/L) dose-dependently increased BEL-7402 cell apoptosis, increased the Bax/Bcl-2 ratio and the levels of cleaved PARP and cleaved caspase-3, and decreased the level of Bcl-2. Furthermore, platycodin D (5-20 μmol/L) induced autophagy in BEL-7402 cells, as evidenced by formation of cytoplasmic vacuoles, increased amounts of LC3-II, and increased numbers of MDC-positive cells. Pretreatment with the autophagy inhibitor chloroquine (5 μmol/L) or BAF (50 nmol/L) significantly enhanced platycodin D-induced proliferation inhibition and apoptosis. Moreover, platycodin D (20 μmol/L) activated the ERK and JNK pathways in BEL-7402 cells, and simultaneous blockage of the two pathways effectively suppressed platycodin D-induced autophagy and enhanced platycodin D-induced apoptosis. In BEL-7402-bearing mice, platycodin D (10 mg·kg(-1)•d(-1)) significantly reduced relative tumor volume with decreased body weight.

Conclusion: Platycodin D not only inhibits the proliferation of BEL-7402 cells but also suppresses BEL-7402 xenograft tumor growth. Platycodin D-induced cell proliferation inhibition and apoptosis are amplified by co-treatment with autophagy inhibitors.

Show MeSH
Related in: MedlinePlus