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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 retards the growth of BEL-7402 xenograft tumors in vivo. After the injection of BEL-7402 cells, mice were treated with different concentrations of PD. The tumor masses taken from tumor-bearing mice were weighed, and the inhibition rate was evaluated (A). The relative tumor volume (B) and body weight (C) were measured. bP<0.05, cP<0.01; eP<0.05, fP<0.01 vs control.
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fig2: PD retards the growth of BEL-7402 xenograft tumors in vivo. After the injection of BEL-7402 cells, mice were treated with different concentrations of PD. The tumor masses taken from tumor-bearing mice were weighed, and the inhibition rate was evaluated (A). The relative tumor volume (B) and body weight (C) were measured. bP<0.05, cP<0.01; eP<0.05, fP<0.01 vs control.

Mentions: We next evaluated the in vivo effect of PD on BEL-7402 xenograft tumor growth. BALB/cA nude mice were subcutaneously injected with BEL-7402 cells and intraperitoneally administered with 10 mg/kg or 5 mg/kg PD for 21 d. The intravenous 5 mg/kg mitomycin C (MMC) group was used as a positive control. As shown in the Figure 2A, a significant decrease (P<0.05) of the tumor weights was detected in mice of the 10 mg/kg PD group compared with the solvent control group. The relative tumor volume in the 10 mg/kg PD-treated group was much lower than that in the solvent control group. After the 21-d treatment, the relative tumor volumes in the 10 mg/kg PD-treated and 5 mg/kg PD-treated groups were 8.10±3.78 and 13.50±5.12, respectively, and the relative tumor volume in the solvent control group was 14.65±4.2 (Figure 2B). In addition to the reduction of body weight after 10 mg/kg PD treatment (Figure 2C), two mice died during the 10 and 5 mg/kg PD treatments, respectively, suggesting that intraperitoneally administered PD had toxic effects in BEL-7402 xenograft mice.


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 retards the growth of BEL-7402 xenograft tumors in vivo. After the injection of BEL-7402 cells, mice were treated with different concentrations of PD. The tumor masses taken from tumor-bearing mice were weighed, and the inhibition rate was evaluated (A). The relative tumor volume (B) and body weight (C) were measured. bP<0.05, cP<0.01; eP<0.05, fP<0.01 vs control.
© Copyright Policy
Related In: Results  -  Collection

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

fig2: PD retards the growth of BEL-7402 xenograft tumors in vivo. After the injection of BEL-7402 cells, mice were treated with different concentrations of PD. The tumor masses taken from tumor-bearing mice were weighed, and the inhibition rate was evaluated (A). The relative tumor volume (B) and body weight (C) were measured. bP<0.05, cP<0.01; eP<0.05, fP<0.01 vs control.
Mentions: We next evaluated the in vivo effect of PD on BEL-7402 xenograft tumor growth. BALB/cA nude mice were subcutaneously injected with BEL-7402 cells and intraperitoneally administered with 10 mg/kg or 5 mg/kg PD for 21 d. The intravenous 5 mg/kg mitomycin C (MMC) group was used as a positive control. As shown in the Figure 2A, a significant decrease (P<0.05) of the tumor weights was detected in mice of the 10 mg/kg PD group compared with the solvent control group. The relative tumor volume in the 10 mg/kg PD-treated group was much lower than that in the solvent control group. After the 21-d treatment, the relative tumor volumes in the 10 mg/kg PD-treated and 5 mg/kg PD-treated groups were 8.10±3.78 and 13.50±5.12, respectively, and the relative tumor volume in the solvent control group was 14.65±4.2 (Figure 2B). In addition to the reduction of body weight after 10 mg/kg PD treatment (Figure 2C), two mice died during the 10 and 5 mg/kg PD treatments, respectively, suggesting that intraperitoneally administered PD had toxic effects in BEL-7402 xenograft mice.

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