Cancer Res Treat.  2011 Mar;43(1):49-55.

Effects of Triterpenoid Glycosides from Fresh Ginseng Berry on SW480 Human Colorectal Cancer Cell Line

• Xie JT
• Du GJ
• McEntee E
• Aung HH
• He H
• Mehendale
• Wang CZ
• Yuan CS

Abstract

PURPOSE
The pharmacological activities, notably the anticancer properties, of bioactive constituents fromfresh American ginseng berry have not yet been well studied. In this study, we investigated the antiproliferative effects of fresh American ginseng berry extract (AGBE) and its representative triterpenoid glycosides using the human colorectal cancer cell line SW480.
MATERIALS AND METHODS
Using high performance liquid chromatography (HPLC), the contents of 8 ginsenosides in AGBE were determined. The cell growth inhibitory effects of AGBE and three triterpenoid glycosides (ginsenosides Rb3, Re, and Rg3) were evaluated by proliferation assay and 3H-thymidine incorporation assay. Cell cycle and apoptotic effects were analyzed by using flow cytometry after staining with propidium iodide and annexin V.
RESULTS
HPLC analysis data showed that AGBE has a distinct ginsenoside profile. AGBE inhibited SW480 cell growth significantly in a time-dependent (24-96 hours) and concentration-dependent (0.1-1.0 mg/mL) manner. Ginsenosides Rb3, Re, and Rg3 also possess significant antiproliferative activities on SW480 cells. 3H-thymidine incorporation assay indicated that AGBE and ginsenosides Rb3, Re, and Rg3 might inhibit the transferring and duplication of DNA in SW480 cells. Flow cytometric assay data suggested that AGBE arrested SW480 cells in S and G2/M phases, and significantly induced cell apoptosis.
CONCLUSION
AGBE and ginsenosides Rb3, Re, and Rg3 possessed significant antiproliferative effects and induced changes of morphological appearance on SW480 cells. The mechanisms of the antiproliferation of AGBE and tested ginsenosides involved could be cell cycle arrest and induction of apoptosis.

Keyword

American ginseng; Ginseng berry; Ginsenoside; SW480; Antiproliferation; Cell cycle; Apoptosis

MeSH Terms

Apoptosis
Cell Cycle
Cell Cycle Checkpoints
Cell Line
Chromatography, High Pressure Liquid
Chromatography, Liquid
Colorectal Neoplasms
DNA
Flow Cytometry
Fruit
Ginsenosides
Glycosides
Humans
Panax
Propidium
DNA
Ginsenosides
Glycosides
Propidium

Figure

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  Fig. 1 High performance liquid chromatographic analyses of American ginseng berry extract (AGBE). (A) Chromatogram of saponin standards, ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd, and Rg3. (B) Chromatogram of AGBE.

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  Fig. 2 Effects of American ginseng berry extract (AGBE) on proliferation of SW480 cells after treatment for different time. SW480 cells were exposed to 1.0 mg/mL of AGBE for 24, 48, 72, and 96 hr. Control at corresponding time is normalized to 100%. *p<0.05, **p<0.01 vs. corresponding control.

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  Fig. 3 Effects of American ginseng berry extract (AGBE) on proliferation and morphological changes of SW480 cells after treatment for 72 hr. (A) SW480 cells were exposed to 0.1, 0.5 and 1.0 mg/mL of AGBE for 72 hr. Control at 72 hr is normalized to 100%. *p<0.05, **p<0.01 vs. control. (B) The morphological aspects of untreated and 1 mg/mL of AGBE treated cells were photographed with a microscope.

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  Fig. 4 Effects of ginsenosides Rb3, Re, and Rg3 on proliferation of SW480 cells. Cells were treated with 0.03, 0.1 and 0.3 mg/mL of ginsenosides for 72 hr. Control at 72 hr is normalized to 100%.

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  Fig. 5 Effects of American ginseng berry extract (AGBE) and ginsenosides on the 3H-thymiding incorporation of SW480 cells. (A) SW480 cells were treated with 0.5 and 1.0 mg/mL of AGBE for 72 hr. (B) SW480 cells were treated with 0.03, 0.1, and 0.3 mg/mL of ginsenosides Rb3, Re, and Rg3 for 72 hr. Control at 72 hr is normalized to 100%. **p<0.01 vs. control.

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  Fig. 6 Cell cycle and apoptosis analysis of SW480 cells using flow cytometry. SW480 cells were treated with 0.5 and 1.0 mg/mL of American ginseng berry extract (AGBE) for 48 hr. (A) Cell cycle profile of SW480 cells. Since not enough viable cells were collected after treatment with 1 mg/mL of AGBE, the cell cycle profile of this group was not calculated. (B) The cytograms of apoptosis assay of SW480 cells. The percentage of apoptotic cells was indicated.

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