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Ginsenoside‑Rg5 induces apoptosis and DNA damage in human cervical cancer cells.

Liang LD, He T, Du TW, Fan YG, Chen DS, Wang Y - Mol Med Rep (2014)

Bottom Line: The HeLa and MS751 cells were significantly more sensitive to ginsenoside‑Rg5 treatment compared with the C‑33A, HT‑3 and Me180 cells.As expected, ginsenoside‑Rg5 induced significant concentration‑ and time‑dependent increases in apoptosis.In addition, ginsenoside‑Rg5 induced significant concentration‑dependent increases in the level of DNA damage compared with the negative control.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.

ABSTRACT
Panax ginseng is traditionally used as a remedy for cancer, inflammation, stress and aging, and ginsenoside‑Rg5 is a major bioactive constituent of steamed ginseng. The present study aimed to evaluate whether ginsenoside‑Rg5 had any marked cytotoxic, apoptotic or DNA‑damaging effects in human cervical cancer cells. Five human cervical cancer cell lines (HeLa, MS751, C33A, Me180 and HT‑3) were used to investigate the cytotoxicity of ginsenoside‑Rg5 using a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. Additionally, the effects of ginsenoside‑Rg5 on the apoptosis of HeLa and MS751 cells were detected using DNA ladder assays and flow cytometry. DNA damage was assessed in the HeLa and MS751 cells using alkaline comet assays and by detection of γH2AX focus formation. The HeLa and MS751 cells were significantly more sensitive to ginsenoside‑Rg5 treatment compared with the C‑33A, HT‑3 and Me180 cells. As expected, ginsenoside‑Rg5 induced significant concentration‑ and time‑dependent increases in apoptosis. In addition, ginsenoside‑Rg5 induced significant concentration‑dependent increases in the level of DNA damage compared with the negative control. Consistent with the comet assay data, the percentage of γH2AX‑positive HeLa and MS751 cells also revealed that ginsenoside‑Rg5 caused DNA double‑strands to break in a concentration‑dependent manner. In conclusion, ginsenoside‑Rg5 had marked genotoxic effects in the HeLa and MS751 cells and, thus, demonstrates potential as a genotoxic or cytotoxic drug for the treatment of cervical cancer.

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DNA double-strand breaks were visualized using γH2AX foci formation in the HeLa and MS751 cells. (A) Images of γH2AX foci formation in cells. Anti-γH2AX monoclonal antibodies were used to detect DNA damage foci immunofluorescence and DAPI was used for nuclei staining (magnification, ×400). (B) Percentages of γH2AX-positive cells were measured following treatment with 10 μM MNNG (CT) and 0, 0.625, 1.25, 2.5 or 5 μM ginsenoside-Rg5 for 24 h. Data are expressed as the mean ± standard error of the mean of three independent experiments. *P<0.05 and **P<0.01, compared with the negative control. CT, positive control; MNNG, N-methyl-N-nitro-N-nitrosoguanidin; DAPI, 4′,6-diamidino-2-phenylindole.
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f5-mmr-11-02-0940: DNA double-strand breaks were visualized using γH2AX foci formation in the HeLa and MS751 cells. (A) Images of γH2AX foci formation in cells. Anti-γH2AX monoclonal antibodies were used to detect DNA damage foci immunofluorescence and DAPI was used for nuclei staining (magnification, ×400). (B) Percentages of γH2AX-positive cells were measured following treatment with 10 μM MNNG (CT) and 0, 0.625, 1.25, 2.5 or 5 μM ginsenoside-Rg5 for 24 h. Data are expressed as the mean ± standard error of the mean of three independent experiments. *P<0.05 and **P<0.01, compared with the negative control. CT, positive control; MNNG, N-methyl-N-nitro-N-nitrosoguanidin; DAPI, 4′,6-diamidino-2-phenylindole.

Mentions: The phosphorylation of histone H2AX foci formation has been suggested as a sensitive way to detect DNA DSBs (27). A threshold of four or more γH2AX foci/cell is optimal for determining the extent of DNA damage (28). Immunofluorescent images of histone H2AX phosphorylation in the γH2AX-stained HeLa and MS751 cells are shown in Fig. 5A. Ginsenoside-Rg5 caused a concentration-dependent induction of γH2AX foci. In the control, the HeLa and MS751 cells had few γH2AX-positive foci in the nuclei and there were ~5.5% cells containing over four foci (Fig. 5B). All treatments with ginsenoside-Rg5 and MNNG induced the formation of foci and increased the percentages of γH2AX-positive cells. In addition, ginsenoside-Rg5 and MNNG exhibited distinct concentration-dependent effects (P<0.01) on the formation of γH2AX foci in the HeLa and MS751 cells (Fig. 5B).


Ginsenoside‑Rg5 induces apoptosis and DNA damage in human cervical cancer cells.

Liang LD, He T, Du TW, Fan YG, Chen DS, Wang Y - Mol Med Rep (2014)

DNA double-strand breaks were visualized using γH2AX foci formation in the HeLa and MS751 cells. (A) Images of γH2AX foci formation in cells. Anti-γH2AX monoclonal antibodies were used to detect DNA damage foci immunofluorescence and DAPI was used for nuclei staining (magnification, ×400). (B) Percentages of γH2AX-positive cells were measured following treatment with 10 μM MNNG (CT) and 0, 0.625, 1.25, 2.5 or 5 μM ginsenoside-Rg5 for 24 h. Data are expressed as the mean ± standard error of the mean of three independent experiments. *P<0.05 and **P<0.01, compared with the negative control. CT, positive control; MNNG, N-methyl-N-nitro-N-nitrosoguanidin; DAPI, 4′,6-diamidino-2-phenylindole.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262516&req=5

f5-mmr-11-02-0940: DNA double-strand breaks were visualized using γH2AX foci formation in the HeLa and MS751 cells. (A) Images of γH2AX foci formation in cells. Anti-γH2AX monoclonal antibodies were used to detect DNA damage foci immunofluorescence and DAPI was used for nuclei staining (magnification, ×400). (B) Percentages of γH2AX-positive cells were measured following treatment with 10 μM MNNG (CT) and 0, 0.625, 1.25, 2.5 or 5 μM ginsenoside-Rg5 for 24 h. Data are expressed as the mean ± standard error of the mean of three independent experiments. *P<0.05 and **P<0.01, compared with the negative control. CT, positive control; MNNG, N-methyl-N-nitro-N-nitrosoguanidin; DAPI, 4′,6-diamidino-2-phenylindole.
Mentions: The phosphorylation of histone H2AX foci formation has been suggested as a sensitive way to detect DNA DSBs (27). A threshold of four or more γH2AX foci/cell is optimal for determining the extent of DNA damage (28). Immunofluorescent images of histone H2AX phosphorylation in the γH2AX-stained HeLa and MS751 cells are shown in Fig. 5A. Ginsenoside-Rg5 caused a concentration-dependent induction of γH2AX foci. In the control, the HeLa and MS751 cells had few γH2AX-positive foci in the nuclei and there were ~5.5% cells containing over four foci (Fig. 5B). All treatments with ginsenoside-Rg5 and MNNG induced the formation of foci and increased the percentages of γH2AX-positive cells. In addition, ginsenoside-Rg5 and MNNG exhibited distinct concentration-dependent effects (P<0.01) on the formation of γH2AX foci in the HeLa and MS751 cells (Fig. 5B).

Bottom Line: The HeLa and MS751 cells were significantly more sensitive to ginsenoside‑Rg5 treatment compared with the C‑33A, HT‑3 and Me180 cells.As expected, ginsenoside‑Rg5 induced significant concentration‑ and time‑dependent increases in apoptosis.In addition, ginsenoside‑Rg5 induced significant concentration‑dependent increases in the level of DNA damage compared with the negative control.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.

ABSTRACT
Panax ginseng is traditionally used as a remedy for cancer, inflammation, stress and aging, and ginsenoside‑Rg5 is a major bioactive constituent of steamed ginseng. The present study aimed to evaluate whether ginsenoside‑Rg5 had any marked cytotoxic, apoptotic or DNA‑damaging effects in human cervical cancer cells. Five human cervical cancer cell lines (HeLa, MS751, C33A, Me180 and HT‑3) were used to investigate the cytotoxicity of ginsenoside‑Rg5 using a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. Additionally, the effects of ginsenoside‑Rg5 on the apoptosis of HeLa and MS751 cells were detected using DNA ladder assays and flow cytometry. DNA damage was assessed in the HeLa and MS751 cells using alkaline comet assays and by detection of γH2AX focus formation. The HeLa and MS751 cells were significantly more sensitive to ginsenoside‑Rg5 treatment compared with the C‑33A, HT‑3 and Me180 cells. As expected, ginsenoside‑Rg5 induced significant concentration‑ and time‑dependent increases in apoptosis. In addition, ginsenoside‑Rg5 induced significant concentration‑dependent increases in the level of DNA damage compared with the negative control. Consistent with the comet assay data, the percentage of γH2AX‑positive HeLa and MS751 cells also revealed that ginsenoside‑Rg5 caused DNA double‑strands to break in a concentration‑dependent manner. In conclusion, ginsenoside‑Rg5 had marked genotoxic effects in the HeLa and MS751 cells and, thus, demonstrates potential as a genotoxic or cytotoxic drug for the treatment of cervical cancer.

Show MeSH
Related in: MedlinePlus