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[Inducement effect of ginsenoside Rg3 on apoptosis of human bladder transitional cell carcinoma cell line EJ]
Jun-xia Chen1, Hui-min Peng, Shu-ping Pu
1Department of Cell Biology and Genetics, Chongqing University of Medical Sciences, Chongqing 400016, China. chjunxia@126.com
Objective:
To explore the effect of Rg3 on inhibiting and inducing apoptosis of bladder cancer cells.
Method:
The bladder cancer cell line EJ was treated with Rg3 of various concentrations. Cell proliferation was measured by MTT assay. Morphological changes of cells were observed by fluorescent staining of Hoechst 33258. Cell cycle and apoptosis rate were analyzed by flow cytometry (FCM). The expression of caspase-3 in cells was detected by immunocytochemistry. DNA ladder was showed by agarose gel electrophoresis.
Result:
Rg3 inhibited proliferation of EJ cells in a manner of concentration-dependent relationship, IC50 of Rg3 in 48 h treatment was 125.5 mg x L(-1) to EJ cells. When treated with 150 mg x L(-1) of Rg3 for 24 h and 48 h, the cells showed apoptotic morphological characteristics including the condensed chromatin, the nuclear fragmentation, the apoptotic body and bright fluorescent granules as well as a higher caspase-3 expression. FCM assay indicated that Rg3 regulated cell cycle and induced apoptosis of EJ cells. When treated for 24 h and 48 h with 75 mg x L(-1) of Rg3 as well as for 48 h with 150 mg x L(-1) of Rg3, the percentages of cells in S phase and G2/M phase were increased, whereas the percentage of cells in G0-G1 was decreased. The apoptosis rates were increased from (1.05 +/- 0.17)% in control group cells to (8.41 +/- 0.98)%, (18.57 +/- 2.20)% and (33.98 +/- 1.64)%, respectively. Remarkable DNA ladders were revealed. The effects showed a manner in dose and time dependent of Rg3.
Conclusion:
The results suggest that ginsenoside Rg3 exerts an inhibiting effect on proliferation of EJ cells by inducing apoptosis.
Insights
Ginsenoside Rg3 effectively inhibits bladder cancer cell proliferation by inducing apoptosis. This compound demonstrated a dose- and time-dependent effect on EJ cells, leading to significant cell cycle regulation and increased apoptosis rates.
Area of Science:
- Oncology
- Pharmacology
- Molecular Biology
Background:
- Bladder cancer remains a significant health concern globally.
- Identifying novel therapeutic agents for bladder cancer is crucial.
- Ginsenoside Rg3, a compound derived from ginseng, has shown potential anti-cancer properties.
Purpose of the Study:
- To investigate the effects of ginsenoside Rg3 on bladder cancer cell proliferation.
- To determine if ginsenoside Rg3 induces apoptosis in bladder cancer cells.
- To analyze the impact of ginsenoside Rg3 on cell cycle progression.
Main Methods:
- EJ bladder cancer cell line was treated with varying concentrations of ginsenoside Rg3.
- Cell proliferation was assessed using MTT assay.
- Apoptosis and cell cycle distribution were analyzed via flow cytometry (FCM).
- Morphological changes were observed using Hoechst 33258 staining, and caspase-3 expression and DNA fragmentation were evaluated.
Main Results:
- Ginsenoside Rg3 exhibited a dose-dependent inhibition of EJ cell proliferation, with an IC50 of 125.5 mg/L at 48 hours.
- Apoptotic morphological characteristics, increased caspase-3 expression, and DNA laddering were observed.
- FCM analysis revealed Rg3 induced apoptosis and altered cell cycle distribution, increasing S and G2/M phases while decreasing G0-G1 phase.
- Apoptosis rates increased significantly, reaching up to 33.98% with specific Rg3 treatments.
Conclusions:
- Ginsenoside Rg3 effectively inhibits bladder cancer cell proliferation.
- The mechanism of inhibition involves the induction of apoptosis.
- The effects of ginsenoside Rg3 are both dose- and time-dependent.
