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20(S)-ginsenoside Rg3 promotes senescence and apoptosis in gallbladder cancer cells via the p53 pathway.

Zhang F, Li M, Wu X, Hu Y, Cao Y, Wang X, Xiang S, Li H, Jiang L, Tan Z, Lu W, Weng H, Shu Y, Gong W, Wang X, Zhang Y, Shi W, Dong P, Gu J, Liu Y - Drug Des Devel Ther (2015)

Bottom Line: However, the anticancer effect of 20(S)-Rg3 in human GBC has not yet been determined.In this study, we primarily found that 20(S)-Rg3 exposure suppressed the survival of both NOZ and GBC-SD cell lines in a concentration-dependent manner.Our results demonstrated that 20(S)-Rg3 potently inhibited growth and survival of GBC cells both in vitro and in vivo. 20(S)-Rg3 attenuated GBC growth probably via activation of the p53 pathway, and subsequent induction of cellular senescence and mitochondrial-dependent apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery and Laboratory of General Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Institute of Biliary Tract Disease, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.

ABSTRACT
Gallbladder cancer (GBC), the most frequent malignancy of the biliary tract, is associated with high mortality and extremely poor prognosis. 20(S)-ginsenoside Rg3 (20(S)-Rg3) is a steroidal saponin with high pharmacological activity. However, the anticancer effect of 20(S)-Rg3 in human GBC has not yet been determined. In this study, we primarily found that 20(S)-Rg3 exposure suppressed the survival of both NOZ and GBC-SD cell lines in a concentration-dependent manner. Moreover, induction of cellular senescence and G0/G1 arrest by 20(S)-Rg3 were accompanied by a large accumulation of p53 and p21 as a result of murine double minute 2 (MDM2) inhibition. 20(S)-Rg3 also caused a remarkable increase in apoptosis via the activation of the mitochondrial-mediated intrinsic caspase pathway. Furthermore, intraperitoneal injection of 20(S)-Rg3 (20 or 40 mg/kg) for 3 weeks markedly inhibited the growth of xenografts in nude mice. Our results demonstrated that 20(S)-Rg3 potently inhibited growth and survival of GBC cells both in vitro and in vivo. 20(S)-Rg3 attenuated GBC growth probably via activation of the p53 pathway, and subsequent induction of cellular senescence and mitochondrial-dependent apoptosis. Therefore, 20(S)-Rg3 may be a potential chemotherapeutic agent for GBC therapy.

No MeSH data available.


Related in: MedlinePlus

20(S)-Rg3 induces apoptosis in gallbladder cancer cells.Notes: (A, B) Changes in apoptotic nuclear morphology were observed by Hoechst 33342 staining and visualized by fluorescent microscopy. (C, D) NOZ and GBC-SD cells were analyzed by flow cytometry with Annexin V-FITC/PI staining after 20(S)-Rg3 treatment. Annexin V vs PI plots from the gated cells showed the populations corresponding to viable (Annexin V−/PI−) and necrotic (Annexin V−/PI+), early (Annexin V+/PI−), and late (Annexin V+/PI+) apoptotic cells. (E, F) Flow cytometric analysis of ΔΨm. NOZ and GBC-SD cells were treated with 20(S)-Rg3 followed by Rhodamine 123 staining. Cells with high ΔΨm are marked “survival” and those with low ΔΨm are marked “apoptosis.” Percentages of cells with low ΔΨm (apoptosis) are shown. (G) Western blot analysis of apoptosis-related proteins in both cell lines. β-actin was used as a loading control. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: SD, standard deviation; FITC, fluorescein isothiocyanate; PI, propidium iodide; vs, versus.
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f4-dddt-9-3969: 20(S)-Rg3 induces apoptosis in gallbladder cancer cells.Notes: (A, B) Changes in apoptotic nuclear morphology were observed by Hoechst 33342 staining and visualized by fluorescent microscopy. (C, D) NOZ and GBC-SD cells were analyzed by flow cytometry with Annexin V-FITC/PI staining after 20(S)-Rg3 treatment. Annexin V vs PI plots from the gated cells showed the populations corresponding to viable (Annexin V−/PI−) and necrotic (Annexin V−/PI+), early (Annexin V+/PI−), and late (Annexin V+/PI+) apoptotic cells. (E, F) Flow cytometric analysis of ΔΨm. NOZ and GBC-SD cells were treated with 20(S)-Rg3 followed by Rhodamine 123 staining. Cells with high ΔΨm are marked “survival” and those with low ΔΨm are marked “apoptosis.” Percentages of cells with low ΔΨm (apoptosis) are shown. (G) Western blot analysis of apoptosis-related proteins in both cell lines. β-actin was used as a loading control. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: SD, standard deviation; FITC, fluorescein isothiocyanate; PI, propidium iodide; vs, versus.

Mentions: We could also see from flow cytometric DNA-histogram that cells were accumulated in the sub-G1 phase following 20(S)-Rg3 treatment. Hoechst 33342 staining was thus performed to examine the nuclear morphology. In control cells, the nuclei were stained weakly, and homogeneously blue, whereas, in cells treated with 20(S)-Rg3 some bright chromatin condensation and nuclear fragmentation were observed (Figure 4A). The numbers of apoptotic nuclei containing condensed chromatin increased significantly as the 20(S)-Rg3 concentration increased (Figure 4B). PI staining also showed a dose-dependent increase in necrosis in both cell lines after 20(S)-Rg3 treatment (Figure S2). A marked dose-dependent increase in both the early and late stages of apoptosis was observed in NOZ and GBC-SC cells following 20(S)-Rg3 treatment by flow cytometry analysis with Annexin V-FITC [fluorescein isothiocyanate]/PI staining (Figure 4C and D). These results from different apoptosis assays reveal significant features of apoptosis, which strongly suggest that 20(S)-Rg3-mediated inhibition of cell growth in GBC is closely correlated with the enhanced apoptosis.


20(S)-ginsenoside Rg3 promotes senescence and apoptosis in gallbladder cancer cells via the p53 pathway.

Zhang F, Li M, Wu X, Hu Y, Cao Y, Wang X, Xiang S, Li H, Jiang L, Tan Z, Lu W, Weng H, Shu Y, Gong W, Wang X, Zhang Y, Shi W, Dong P, Gu J, Liu Y - Drug Des Devel Ther (2015)

20(S)-Rg3 induces apoptosis in gallbladder cancer cells.Notes: (A, B) Changes in apoptotic nuclear morphology were observed by Hoechst 33342 staining and visualized by fluorescent microscopy. (C, D) NOZ and GBC-SD cells were analyzed by flow cytometry with Annexin V-FITC/PI staining after 20(S)-Rg3 treatment. Annexin V vs PI plots from the gated cells showed the populations corresponding to viable (Annexin V−/PI−) and necrotic (Annexin V−/PI+), early (Annexin V+/PI−), and late (Annexin V+/PI+) apoptotic cells. (E, F) Flow cytometric analysis of ΔΨm. NOZ and GBC-SD cells were treated with 20(S)-Rg3 followed by Rhodamine 123 staining. Cells with high ΔΨm are marked “survival” and those with low ΔΨm are marked “apoptosis.” Percentages of cells with low ΔΨm (apoptosis) are shown. (G) Western blot analysis of apoptosis-related proteins in both cell lines. β-actin was used as a loading control. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: SD, standard deviation; FITC, fluorescein isothiocyanate; PI, propidium iodide; vs, versus.
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Related In: Results  -  Collection

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f4-dddt-9-3969: 20(S)-Rg3 induces apoptosis in gallbladder cancer cells.Notes: (A, B) Changes in apoptotic nuclear morphology were observed by Hoechst 33342 staining and visualized by fluorescent microscopy. (C, D) NOZ and GBC-SD cells were analyzed by flow cytometry with Annexin V-FITC/PI staining after 20(S)-Rg3 treatment. Annexin V vs PI plots from the gated cells showed the populations corresponding to viable (Annexin V−/PI−) and necrotic (Annexin V−/PI+), early (Annexin V+/PI−), and late (Annexin V+/PI+) apoptotic cells. (E, F) Flow cytometric analysis of ΔΨm. NOZ and GBC-SD cells were treated with 20(S)-Rg3 followed by Rhodamine 123 staining. Cells with high ΔΨm are marked “survival” and those with low ΔΨm are marked “apoptosis.” Percentages of cells with low ΔΨm (apoptosis) are shown. (G) Western blot analysis of apoptosis-related proteins in both cell lines. β-actin was used as a loading control. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: SD, standard deviation; FITC, fluorescein isothiocyanate; PI, propidium iodide; vs, versus.
Mentions: We could also see from flow cytometric DNA-histogram that cells were accumulated in the sub-G1 phase following 20(S)-Rg3 treatment. Hoechst 33342 staining was thus performed to examine the nuclear morphology. In control cells, the nuclei were stained weakly, and homogeneously blue, whereas, in cells treated with 20(S)-Rg3 some bright chromatin condensation and nuclear fragmentation were observed (Figure 4A). The numbers of apoptotic nuclei containing condensed chromatin increased significantly as the 20(S)-Rg3 concentration increased (Figure 4B). PI staining also showed a dose-dependent increase in necrosis in both cell lines after 20(S)-Rg3 treatment (Figure S2). A marked dose-dependent increase in both the early and late stages of apoptosis was observed in NOZ and GBC-SC cells following 20(S)-Rg3 treatment by flow cytometry analysis with Annexin V-FITC [fluorescein isothiocyanate]/PI staining (Figure 4C and D). These results from different apoptosis assays reveal significant features of apoptosis, which strongly suggest that 20(S)-Rg3-mediated inhibition of cell growth in GBC is closely correlated with the enhanced apoptosis.

Bottom Line: However, the anticancer effect of 20(S)-Rg3 in human GBC has not yet been determined.In this study, we primarily found that 20(S)-Rg3 exposure suppressed the survival of both NOZ and GBC-SD cell lines in a concentration-dependent manner.Our results demonstrated that 20(S)-Rg3 potently inhibited growth and survival of GBC cells both in vitro and in vivo. 20(S)-Rg3 attenuated GBC growth probably via activation of the p53 pathway, and subsequent induction of cellular senescence and mitochondrial-dependent apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery and Laboratory of General Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Institute of Biliary Tract Disease, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.

ABSTRACT
Gallbladder cancer (GBC), the most frequent malignancy of the biliary tract, is associated with high mortality and extremely poor prognosis. 20(S)-ginsenoside Rg3 (20(S)-Rg3) is a steroidal saponin with high pharmacological activity. However, the anticancer effect of 20(S)-Rg3 in human GBC has not yet been determined. In this study, we primarily found that 20(S)-Rg3 exposure suppressed the survival of both NOZ and GBC-SD cell lines in a concentration-dependent manner. Moreover, induction of cellular senescence and G0/G1 arrest by 20(S)-Rg3 were accompanied by a large accumulation of p53 and p21 as a result of murine double minute 2 (MDM2) inhibition. 20(S)-Rg3 also caused a remarkable increase in apoptosis via the activation of the mitochondrial-mediated intrinsic caspase pathway. Furthermore, intraperitoneal injection of 20(S)-Rg3 (20 or 40 mg/kg) for 3 weeks markedly inhibited the growth of xenografts in nude mice. Our results demonstrated that 20(S)-Rg3 potently inhibited growth and survival of GBC cells both in vitro and in vivo. 20(S)-Rg3 attenuated GBC growth probably via activation of the p53 pathway, and subsequent induction of cellular senescence and mitochondrial-dependent apoptosis. Therefore, 20(S)-Rg3 may be a potential chemotherapeutic agent for GBC therapy.

No MeSH data available.


Related in: MedlinePlus