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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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Ginsenoside-Rg5 induces apoptosis in HeLa and MS751 cells. (A) Apoptosis-associated DNA fragmentation in HeLa and MS751 cells. (B) Apoptotic rates of HeLa and MS751 cells treated with ginsenoside-Rg5 (0–5 μM) and MNNG (10 μM) 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, vs. negative control (0 μM ginsenoside-Rg5). P-values refer to the comparison of baseline independent characteristics. (C) Flow cytometric analysis of apoptosis using Annexin-V and PI double-staining. MNNG, N-methyl-N-nitro-N-nitrosoguanidin; M, marker of DNA ladder; CT, positive control (10 μM MNNG); PI, propidium iodide.
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f3-mmr-11-02-0940: Ginsenoside-Rg5 induces apoptosis in HeLa and MS751 cells. (A) Apoptosis-associated DNA fragmentation in HeLa and MS751 cells. (B) Apoptotic rates of HeLa and MS751 cells treated with ginsenoside-Rg5 (0–5 μM) and MNNG (10 μM) 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, vs. negative control (0 μM ginsenoside-Rg5). P-values refer to the comparison of baseline independent characteristics. (C) Flow cytometric analysis of apoptosis using Annexin-V and PI double-staining. MNNG, N-methyl-N-nitro-N-nitrosoguanidin; M, marker of DNA ladder; CT, positive control (10 μM MNNG); PI, propidium iodide.

Mentions: The HeLa and MS751 cells were significantly more sensitive to ginsenoside-Rg5. Therefore, to determine whether the loss in cell viability induced by ginsenoside-Rg5 was associated with apoptosis, ginsenoside-Rg5-induced apoptosis was examined in these sensitive cell lines using DNA fragmentation analysis and flow cytometry. As shown in Fig. 3A, ginsenoside-Rg5 induced a ladder-like pattern on the urea polyacrylamide gel electrophoresis (PAGE). The DNA in the untreated cells remained intact. At concentrations of 2.5 and 5 μM, ginsenoside-Rg5 led to concentration- and time-dependent increases in DNA fragmentation. In addition, in HeLa and MS751 cells, the fraction of apoptotic cells increased markedly when the cells were treated with 1.25–5 μM ginsenoside-Rg5 (Fig. 3B). Taken together, these results suggested that the anticancer activity of ginsenoside-Rg5 was mediated, in part, by the induction of apoptosis.


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)

Ginsenoside-Rg5 induces apoptosis in HeLa and MS751 cells. (A) Apoptosis-associated DNA fragmentation in HeLa and MS751 cells. (B) Apoptotic rates of HeLa and MS751 cells treated with ginsenoside-Rg5 (0–5 μM) and MNNG (10 μM) 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, vs. negative control (0 μM ginsenoside-Rg5). P-values refer to the comparison of baseline independent characteristics. (C) Flow cytometric analysis of apoptosis using Annexin-V and PI double-staining. MNNG, N-methyl-N-nitro-N-nitrosoguanidin; M, marker of DNA ladder; CT, positive control (10 μM MNNG); PI, propidium iodide.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-mmr-11-02-0940: Ginsenoside-Rg5 induces apoptosis in HeLa and MS751 cells. (A) Apoptosis-associated DNA fragmentation in HeLa and MS751 cells. (B) Apoptotic rates of HeLa and MS751 cells treated with ginsenoside-Rg5 (0–5 μM) and MNNG (10 μM) 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, vs. negative control (0 μM ginsenoside-Rg5). P-values refer to the comparison of baseline independent characteristics. (C) Flow cytometric analysis of apoptosis using Annexin-V and PI double-staining. MNNG, N-methyl-N-nitro-N-nitrosoguanidin; M, marker of DNA ladder; CT, positive control (10 μM MNNG); PI, propidium iodide.
Mentions: The HeLa and MS751 cells were significantly more sensitive to ginsenoside-Rg5. Therefore, to determine whether the loss in cell viability induced by ginsenoside-Rg5 was associated with apoptosis, ginsenoside-Rg5-induced apoptosis was examined in these sensitive cell lines using DNA fragmentation analysis and flow cytometry. As shown in Fig. 3A, ginsenoside-Rg5 induced a ladder-like pattern on the urea polyacrylamide gel electrophoresis (PAGE). The DNA in the untreated cells remained intact. At concentrations of 2.5 and 5 μM, ginsenoside-Rg5 led to concentration- and time-dependent increases in DNA fragmentation. In addition, in HeLa and MS751 cells, the fraction of apoptotic cells increased markedly when the cells were treated with 1.25–5 μM ginsenoside-Rg5 (Fig. 3B). Taken together, these results suggested that the anticancer activity of ginsenoside-Rg5 was mediated, in part, by the induction of apoptosis.

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