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

Liang,Li‑Dan; He,Tao; Du,Ting‑Wei; Fan,Yong‑Gang; Chen,Dian‑Sen; Wang,Yan

doi:10.3892/mmr.2014.2821

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

Authors:
- Li‑Dan Liang
- Tao He
- Ting‑Wei Du
- Yong‑Gang Fan
- Dian‑Sen Chen
- Yan Wang
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China
Published online on: October 30, 2014 https://doi.org/10.3892/mmr.2014.2821
Pages: 940-946
Copyright: © Liang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

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.

View Figures

View References

1	Saslow D, Castle PE, Cox JT, et al: American Cancer Society. Guideline for human papillomavirus (HPV) vaccine use to prevent cervical cancer and its precursors. CA Cancer J Clin. 57:7–28. 2007. View Article : Google Scholar : PubMed/NCBI
2	Steben M and Duarte-Franco E: Human papillomavirus infection: epidemiology and pathophysiology. Gynecol Oncol. 107:S2–S5. 2007. View Article : Google Scholar : PubMed/NCBI
3	Long HJ 3rd: Management of metastatic cervical cancer: review of the literature. J Clin Oncol. 25:2966–2974. 2007. View Article : Google Scholar : PubMed/NCBI
4	Brinkman JA, Caffrey AS, Muderspach LI, Roman LD and Kast WM: Inhibitory effect of ginsenoside Rg5 and its metabolite ginsenoside Rh3 in an oxazolone induced mouse chronic dermatitis model. Eur J Gynaecol Oncol. 26:129–142. 2005.
5	Leitao MM Jr and Chi DS: Recurrent cervical cancer. Curr Treat Options Oncol. 3:105–111. 2002. View Article : Google Scholar : PubMed/NCBI
6	Yang J, Li J, Sun M and Chen K: Studies of traditional Chinese medicine monomer on HeLa cell of cervical cancer. Pak J Pharm Sci. 27(4 Suppl): 1063–1068. 2014.PubMed/NCBI
7	Cohen I, Tagliaferri M and Tripathy D: Traditional Chinese medicine in the treatment of breast cancer. Semin Oncol. 29:563–574. 2002. View Article : Google Scholar
8	Li X, Yang G, Li X, et al: Traditional Chinese medicine in cancer care: a review of controlled clinical studies published in Chinese. PloS One. 8:e603382013. View Article : Google Scholar : PubMed/NCBI
9	Yang G, Li X, Li X, et al: Traditional chinese medicine in cancer care: a review of case series published in the chinese literature. Evid Based Complement Alternat Med. 2012.7510–7546. 2012.
10	Liu J, Li X, Liu J, Ma L, Li X and Fønnebø V: Traditional Chinese medicine in cancer care: a review of case reports published in Chinese literature. Forsch Komplementarmed. 18:257–263. 2011. View Article : Google Scholar
11	Kwon SW, Han SB, Park IH, Kim JM, Park MK and Park JH: Liquid chromatographic determination of less polar ginsenosides in processed ginseng. J Chromatogr A. 921:335–339. 2001. View Article : Google Scholar : PubMed/NCBI
12	Kim SN, Ha YW, Shin H, Son SH, Wu SJ and Kim YS: Simultaneous quantification of 14 ginsenosides in Panax ginseng C.A. Meyer (Korean red ginseng) by HPLC-ELSD and its application to quality control. J Pharm Biomed Anal. 45:164–170. 2007. View Article : Google Scholar : PubMed/NCBI
13	Yun TK, Lee YS, Lee YH, Kim SI and Yun HY: Anticarcino genic effect of Panax ginseng C.A. Meyer and identification of active compounds. J Korean Med Sci. 16:S6–S18. 2001. View Article : Google Scholar
14	Kang KS, Kim HY, Baek SH, et al: Study on the hydroxyl radical scavenging activity changes of ginseng and ginsenoside-Rb2 by heat processing. Biol Pharm Bull. 30:724–728. 2007. View Article : Google Scholar : PubMed/NCBI
15	Nag SA, Qin JJ, Wang W, et al: Ginsenosides as anticancer agents: In vitro and in vivo activities, structure-activity relationships, and molecular mechanisms of action. Front Pharmacol. 3:252012. View Article : Google Scholar : PubMed/NCBI
16	Lee KY, Lee YH, Kim SI, Park JH and Lee SK: Ginsenoside-Rg5 suppresses cyclin E-dependent protein kinase activity via up-regulating p21Cip/WAF1 and down-regulating cyclin E in SK-HEP-1 cells. Anticancer Res. 17:1067–1072. 1997.PubMed/NCBI
17	Shin YW, Bae EA and Kim DH: Inhibitory effect of ginsenoside Rg5 and its metabolite ginsenoside Rh3 in an oxazolone-induced mouse chronic dermatitis model. Arch Pharm Res. 29:685–690. 2006. View Article : Google Scholar : PubMed/NCBI
18	Kim TW, Joh EH, Kim B and Kim DH: Ginsenoside Rg5 ameliorates lung inflammation in mice by inhibiting the binding of LPS to toll-like receptor-4 on macrophages. Int Immunopharmacol. 12:110–116. 2012. View Article : Google Scholar
19	Kim EJ, Jung IH, Van Le TK, Jeong JJ, Kim NJ and Kim DH: Ginsenosides Rg5 and Rh3 protect scopolamine-induced memory deficits in mice. J Ethnopharmacol. 146:294–299. 2013. View Article : Google Scholar : PubMed/NCBI
20	Lee YY, Park JS, Jung JS, Kim DH and Kim HS: Anti-inflammatory effect of ginsenoside Rg5 in lipopolysaccharide-stimulated BV2 microglial cells. Int J Mol Sci. 14:9820–9833. 2013. View Article : Google Scholar : PubMed/NCBI
21	Kawai K, Liu SX, Tyurin VA, et al: Antioxidant and antiapoptotic function of metallothioneins in HL-60 cells challenged with copper nitrilotriacetate. Chem Res Toxicol. 13:1275–1286. 2000. View Article : Google Scholar : PubMed/NCBI
22	Wu SB, Su JJ, Sun LH, et al: Triterpenoids and steroids from the fruits of Melia toosendan and their cytotoxic effects on two human cancer cell lines. J Natl Prod. 73:1898–1906. 2010. View Article : Google Scholar
23	Nakadai A, Li Q and Kawada T: Chlorpyrifos induces apoptosis in human monocyte cell line U937. Toxicology. 224:202–209. 2006. View Article : Google Scholar : PubMed/NCBI
24	Guanggang X, Diqiu L, Jianzhong Y, et al: Carbamate insecticide methomyl confers cytotoxicity through DNA damage induction. Food Chem Toxicol. 53:352–358. 2013. View Article : Google Scholar
25	Zhang YH, Li HD, Li B, Jiang SD and Jiang LS: Ginsenoside Rg3 induces DNA damage in human osteosarcoma cells and reduces MNNG-induced DNA damage and apoptosis in normal human cells. Oncol Rep. 31:919–925. 2014.
26	Wu GS, Lu JJ, Guo JJ, et al: Ganoderic acid DM, a natural triterpenoid, induces DNA damage, G1 cell cycle arrest and apoptosis in human breast cancer cells. Fitoterapia. 83:408–414. 2012. View Article : Google Scholar
27	Sokolov MV, Smilenov LB, Hall EJ, Panyutin IG, Bonner WM and Sedelnikova OA: Ionizing radiation induces DNA double-strand breaks in bystander primary human fibroblasts. Oncogene. 24:7257–7265. 2005. View Article : Google Scholar : PubMed/NCBI
28	Parry MC, Bhabra G, Sood A, et al: Thresholds for indirect DNA damage across cellular barriers for orthopaedic biomaterials. Biomaterials. 31:4477–4483. 2010. View Article : Google Scholar : PubMed/NCBI
29	Toh DF, Patel DN, Chan EC, Teo A, Neo SY and Koh HL: Anti-proliferative effects of raw and steamed extracts of Panax notoginseng and its ginsenoside constituents on human liver cancer cells. Chin Med. 6:42011. View Article : Google Scholar : PubMed/NCBI
30	Yue PY, Mak NK, Cheng YK, et al: Pharmacogenomics and the Yin/Yang actions of ginseng: anti-tumor, angiomodulating and steroid-like activities of ginsenosides. Chin Med. 2:62007. View Article : Google Scholar : PubMed/NCBI
31	Bras M, Queenan B and Susin SA: Programmed cell death via mitochondria: different modes of dying. Biochemistry (Mosc). 70:231–239. 2005. View Article : Google Scholar
32	Broker LE, Kruyt FA and Giaccone G: Cell death independent of caspases: a review. Clin Cancer Res. 11:3155–3162. 2005. View Article : Google Scholar : PubMed/NCBI
33	Edinger AL and Thompson CB: Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol. 16:663–669. 2004. View Article : Google Scholar : PubMed/NCBI
34	Kim HS, Lee EH, Ko SR, Choi KJ, Park JH and Im DS: Effects of ginsenosides Rg3 and Rh2 on the proliferation of prostate cancer cells. Arch Pharm Res. 27:429–435. 2004. View Article : Google Scholar : PubMed/NCBI
35	Park HM, Kim SJ, Kim JS and Kang HS: Reactive oxygen species mediated ginsenoside Rg3- and Rh2-induced apoptosis in hepatoma cells through mitochondrial signaling pathways. Food Chem Toxicol. 50:2736–2741. 2012. View Article : Google Scholar : PubMed/NCBI
36	Li B, Zhao J, Wang CZ, et al: Ginsenoside Rh2 induces apoptosis and paraptosis-like cell death in colorectal cancer cells through activation of p53. Cancer Lett. 301:185–192. 2011. View Article : Google Scholar : PubMed/NCBI
37	Kim SE, Lee YH, Park JH and Lee SK: Ginsenoside-Rs4, a new type of ginseng saponin concurrently induces apoptosis and selectively elevates protein levels of p53 and p21WAF1 in human hepatoma SK-HEP-1 cells. Eur J Cancer. 35:507–511. 1999. View Article : Google Scholar : PubMed/NCBI
38	Oh SH and Lee BH: A ginseng saponin metabolite-induced apoptosis in HepG2 cells involves a mitochondria-mediated pathway and its downstream caspase-8 activation and Bid cleavage. Toxicol Appl Pharmacol. 194:221–229. 2004. View Article : Google Scholar : PubMed/NCBI
39	Robertson JD and Orrenius S: Role of mitochondria in toxic cell death. Toxicology. 181–182:491–496. 2002. View Article : Google Scholar
40	Poon PY, Kwok HH, Yue PY, et al: Cytoprotective effect of 20S-Rg3 on benzo[a]pyrene-induced DNA damage. Drug Metab Dispos. 40:120–129. 2012. View Article : Google Scholar
41	Cotelle S and Férard JF: Comet assay in genetic ecotoxicology: a review. Environ Mol Mutagen. 34:246–255. 1999. View Article : Google Scholar
42	Yu Y, Zhu W, Diao H, Zhou C, Chen FF and Yang J: A comparative study of using comet assay and gammaH2AX foci formation in the detection of N-methyl-N′-nitro-N-nitrosoguanidine-induced DNA damage. Toxicology In Vitro. 20:959–965. 2006. View Article : Google Scholar