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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 represses tumor growth in xenografted nude mice model by causing apoptotic cell death.Notes: Tumor xenografts were established by subcutaneous inoculation of NOZ cells into the right flank of nude mice. (A) The mice were then administered 0.2 mL of vehicle (PBS) or 20(S)-Rg3 (20 and 40 mg/kg) IP every day for up to 3 weeks. (B) Tumor dimensions were periodically measured using calipers. (C, D) Photographs of representative tumors from each group are shown. Tumors were excised from the animals and weighed. (E) Survival curves of the mice in control and 20(S)-Rg3 (40 mg/kg) groups are shown. (F) SA-β-gal staining showed that 20(S)-Rg3 induced more senescent cells in tumor tissues. Vacuolization by 20(S)-Rg3 was accompanied by a large accumulation of p53 and p21 in tumor tissues. (G, H) Immunofluorescent images illustrate location of PCNA and cleaved-caspase-3 proteins in tumor tissues. Green fluorescence shows nuclear expression of PCNA, red fluorescence shows cytoplasmic expression of cleaved-caspase-3, and blue fluorescence shows all cell nuclei with patterns of DAPI staining. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; PCNA, proliferating cell nuclear antigen; PBS, phosphate buffered saline; IP, intraperitoneal; SD, standard deviation; vs, versus; SA-β-gal, senescence-associated β-galactosidase.
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f6-dddt-9-3969: 20(S)-Rg3 represses tumor growth in xenografted nude mice model by causing apoptotic cell death.Notes: Tumor xenografts were established by subcutaneous inoculation of NOZ cells into the right flank of nude mice. (A) The mice were then administered 0.2 mL of vehicle (PBS) or 20(S)-Rg3 (20 and 40 mg/kg) IP every day for up to 3 weeks. (B) Tumor dimensions were periodically measured using calipers. (C, D) Photographs of representative tumors from each group are shown. Tumors were excised from the animals and weighed. (E) Survival curves of the mice in control and 20(S)-Rg3 (40 mg/kg) groups are shown. (F) SA-β-gal staining showed that 20(S)-Rg3 induced more senescent cells in tumor tissues. Vacuolization by 20(S)-Rg3 was accompanied by a large accumulation of p53 and p21 in tumor tissues. (G, H) Immunofluorescent images illustrate location of PCNA and cleaved-caspase-3 proteins in tumor tissues. Green fluorescence shows nuclear expression of PCNA, red fluorescence shows cytoplasmic expression of cleaved-caspase-3, and blue fluorescence shows all cell nuclei with patterns of DAPI staining. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; PCNA, proliferating cell nuclear antigen; PBS, phosphate buffered saline; IP, intraperitoneal; SD, standard deviation; vs, versus; SA-β-gal, senescence-associated β-galactosidase.

Mentions: To further evaluate whether 20(S)-Rg3 had an effect on inhibition of tumor growth in vivo, we measured the tumor volume in a xenograft tumor model in which NOZ cells were injected IP into nude mice. As shown in Figure 6A, we found that the drug concentration in the serum of mice with 40 mg/kg 20(S)-Rg3 administered IP daily for 1 week reached a peak at about 200 μM by a HPLC assay, which just fell within the range of the drug concentrations (25–400 μM) in vitro. So, 20 and 40 mg/kg of 20(S)-Rg3 were selected for in vivo experiments. When transplant tumors reached a mean group size of approximately 100 mm3, mice were treated every day for 3 weeks with various dose of 20(S)-Rg3. 20(S)-Rg3 showed a significant inhibitory effect on tumor volume and weight (Figure 6B–D). The survival of xenografted nude mice was prolonged after 40 mg/kg 20(S)-Rg3 treatment (Figure 6E). Moreover, SA-β-gal staining of tumor sections indicated that 20(S)-Rg3 caused tumor cell senescence. 20(S)-Rg3 dose-dependently induced a significant increase in p53 and p21Waf1 expression (Figure 6F). In addition, tumor cell density and nuclear amount were both reduced. These results indicate that 20(S)-Rg3 inhibit tumor growth by causing cell senescence and apoptosis. Furthermore, PCNA expression was decreased, and caspase-3 activity was increased in 20(S)-Rg3 treatment groups (Figure 6G and H).


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 represses tumor growth in xenografted nude mice model by causing apoptotic cell death.Notes: Tumor xenografts were established by subcutaneous inoculation of NOZ cells into the right flank of nude mice. (A) The mice were then administered 0.2 mL of vehicle (PBS) or 20(S)-Rg3 (20 and 40 mg/kg) IP every day for up to 3 weeks. (B) Tumor dimensions were periodically measured using calipers. (C, D) Photographs of representative tumors from each group are shown. Tumors were excised from the animals and weighed. (E) Survival curves of the mice in control and 20(S)-Rg3 (40 mg/kg) groups are shown. (F) SA-β-gal staining showed that 20(S)-Rg3 induced more senescent cells in tumor tissues. Vacuolization by 20(S)-Rg3 was accompanied by a large accumulation of p53 and p21 in tumor tissues. (G, H) Immunofluorescent images illustrate location of PCNA and cleaved-caspase-3 proteins in tumor tissues. Green fluorescence shows nuclear expression of PCNA, red fluorescence shows cytoplasmic expression of cleaved-caspase-3, and blue fluorescence shows all cell nuclei with patterns of DAPI staining. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; PCNA, proliferating cell nuclear antigen; PBS, phosphate buffered saline; IP, intraperitoneal; SD, standard deviation; vs, versus; SA-β-gal, senescence-associated β-galactosidase.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4539091&req=5

f6-dddt-9-3969: 20(S)-Rg3 represses tumor growth in xenografted nude mice model by causing apoptotic cell death.Notes: Tumor xenografts were established by subcutaneous inoculation of NOZ cells into the right flank of nude mice. (A) The mice were then administered 0.2 mL of vehicle (PBS) or 20(S)-Rg3 (20 and 40 mg/kg) IP every day for up to 3 weeks. (B) Tumor dimensions were periodically measured using calipers. (C, D) Photographs of representative tumors from each group are shown. Tumors were excised from the animals and weighed. (E) Survival curves of the mice in control and 20(S)-Rg3 (40 mg/kg) groups are shown. (F) SA-β-gal staining showed that 20(S)-Rg3 induced more senescent cells in tumor tissues. Vacuolization by 20(S)-Rg3 was accompanied by a large accumulation of p53 and p21 in tumor tissues. (G, H) Immunofluorescent images illustrate location of PCNA and cleaved-caspase-3 proteins in tumor tissues. Green fluorescence shows nuclear expression of PCNA, red fluorescence shows cytoplasmic expression of cleaved-caspase-3, and blue fluorescence shows all cell nuclei with patterns of DAPI staining. Data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 vs control.Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; PCNA, proliferating cell nuclear antigen; PBS, phosphate buffered saline; IP, intraperitoneal; SD, standard deviation; vs, versus; SA-β-gal, senescence-associated β-galactosidase.
Mentions: To further evaluate whether 20(S)-Rg3 had an effect on inhibition of tumor growth in vivo, we measured the tumor volume in a xenograft tumor model in which NOZ cells were injected IP into nude mice. As shown in Figure 6A, we found that the drug concentration in the serum of mice with 40 mg/kg 20(S)-Rg3 administered IP daily for 1 week reached a peak at about 200 μM by a HPLC assay, which just fell within the range of the drug concentrations (25–400 μM) in vitro. So, 20 and 40 mg/kg of 20(S)-Rg3 were selected for in vivo experiments. When transplant tumors reached a mean group size of approximately 100 mm3, mice were treated every day for 3 weeks with various dose of 20(S)-Rg3. 20(S)-Rg3 showed a significant inhibitory effect on tumor volume and weight (Figure 6B–D). The survival of xenografted nude mice was prolonged after 40 mg/kg 20(S)-Rg3 treatment (Figure 6E). Moreover, SA-β-gal staining of tumor sections indicated that 20(S)-Rg3 caused tumor cell senescence. 20(S)-Rg3 dose-dependently induced a significant increase in p53 and p21Waf1 expression (Figure 6F). In addition, tumor cell density and nuclear amount were both reduced. These results indicate that 20(S)-Rg3 inhibit tumor growth by causing cell senescence and apoptosis. Furthermore, PCNA expression was decreased, and caspase-3 activity was increased in 20(S)-Rg3 treatment groups (Figure 6G and H).

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