Effects of ginsenoside Rg2 on the ultraviolet B-induced DNA damage responses in HaCaT cells

  • Se Eun Ha
  • Dae Hyun Shin
  • Hyung Do Kim
  • Sun Mi Shim
  • Hack Soo Kim
  • Bo Hyeon Kim
  • Jung Sup Lee
  • Jong Kun ParkEmail author


Our previous study demonstrated the increase in the repair of UVB damage by mRg2, a mixture of ginsenosides containing 60% Rg2 in NIH3T3 cells. In the present study, the effects of purified Rg2 on the repair and apoptosis in ultraviolet B (UVB)-exposed HaCaT cells were investigated on gene expression levels. When cells were exposed to UVB and post-incubated in normal medium for 24 h, the cell viability decreased to about 50% of that in nontreated control. When Rg2 was post-incubated, however, the UVB-induced cytotoxicity was significantly prevented in an Rg2 concentration- and time-dependent manner. The apoptotic nuclear fragmentation resulting from UVB exposure was also significantly protected by the Rg2 post-incubation. Microarray analysis showed that the genes stimulated by the Rg2-alone treatment include those involved in p53 signaling pathway such as GADD45α, GADD45β, and cell communication genes. RT-PCR analysis showed that the Rg2-alone treatment slightly upregulated the p53 and GADD45 transcript and protein levels by about 1.5-fold as compared with the nontreated control. The mRNA levels of p53 and GADD45 in cells exposed to UVB and post-incubated with Rg2 for 24 h decreased in an Rg2 concentration-dependent manner as compared with that post-incubated in normal medium. However, the mRNA level of the UVB-exposed cells post-incubated with 5 μM retinol was essentially the same as that post-incubated in normal medium. Time course experiment showed that the mRNA levels of p53 and GADD45 in UVB-exposed cells were upregulated by post-incubation with 50 μM Rg2 until 6 and 9 h, respectively, and then gradually decreased until 24 h. By Western blot analysis, it was also revealed that the Rg2 post-incubation decreases the expression of p53, phospho-p53, GADD45, and ATM in UVB-exposed cells. Time course analysis also indicated that these decreased expressions were due to the earlier upregulation of p53 and GADD45 proteins. When UVB-exposed cells were post-incubated with Rg2 for 24 h after UVB exposure, the level of remaining cyclobutane pyrimidine dimers decreased in both Rg2 concentration- and time-dependent manner. All these results suggest that Rg2 protects cells against UVB-induced genotoxicity by increasing DNA repair, in possible association with modulation of protein levels involved in p53 signaling pathway.


Ginsenoside Rg2 UVB Repair Apoptosis p53 GADD45 



The present study was supported by SMBA (2008-JB-SMBA). Dae Hyun Shin, Hack Soo Kim, and Bo Hyeon Kim were supported by Somang Cosmetics, Korea.


  1. Afaq F, Adhami VM, Ahmad N, Mukhtar H (2002) Botanical antioxidants for chemoprevention of photocarcinogenesis. Front Biosci 7:d784–d792CrossRefPubMedGoogle Scholar
  2. Apone F, Tito A, Carola A, Arciello S, Tortora A, Filippini L, Monoli I, Cucchiara M, Gibertoni S, Chrispeels MJ, Colucci G (2010) A mixture of peptides and sugars derived from plant cell walls increases plant defense responses to stress and attenuates ageing-associated molecular changes in cultured skin cells. J Biotech 145:367–376CrossRefGoogle Scholar
  3. Bachelor MA, Bowden GT (2004) UVA-mediated activation of signaling pathways involved in skin tumor promotion and progression. Semin Cancer Biol 14(2):131–138CrossRefPubMedGoogle Scholar
  4. Böhm M, Wolff I, Scholzen TE, Robinson SJ, Healy E, Luger TA, Schwarz T, Schwarz A (2005) alpha-Melanocyte-stimulating hormone protects from ultraviolet radiation-induced apoptosis and DNA damage. J Biol Chem 280:5795–5802CrossRefPubMedGoogle Scholar
  5. Cai BX, Luo D, Lin XF, Gao J (2008) Compound K suppresses ultraviolet radiation-induced apoptosis by inducing DNA repair in human keratinocytes. Arch Pharm Res 31(11):1483–1488CrossRefPubMedGoogle Scholar
  6. Chang L, Zhou B, Hu S, Guo R, Liu X, Jones SN, Yen Y (2008) ATM-mediated serine 72 phosphorylation stabilizes ribonucleotide reductase small subunit p53R2 protein against MDM2 to DNA damage. Proc Natl Acad Sci U S A 105(47):18519–18524CrossRefPubMedGoogle Scholar
  7. Cho SW, Cho EH, Choi SY (1995) Ginsenosides activate DNA polymerase delta from bovine placenta. Life Sci 57(14):1359–1365CrossRefPubMedGoogle Scholar
  8. Clem RJ, Miller LK (1994) Prevention of apoptosis by a baculovirus gene during infection of insect cells. Mol Cell Biol 14:5212–5222PubMedGoogle Scholar
  9. Cuesta A, Zambrano A, Royo M, Pascual A (2008) The tumor suppressor p53 regulates the expression of the beta-amyloid precursor protein (APP). Biochem J 418(3):643–650CrossRefGoogle Scholar
  10. Curtin JC, Dragnev KH, Sekula D, Christie AJ, Dmitrovsky E, Spinella MJ (2001) Retinoic acid activates p53 in human embryonal carcinoma through retinoid receptor-dependent stimulation of p53 transactivation function. Oncogene 20(20):2559–2569CrossRefPubMedGoogle Scholar
  11. De Gruijl FR (1999) Skin cancer and solar UV radiation. Eur J Cancer 35:2003–2009CrossRefPubMedGoogle Scholar
  12. Dregoesc D, Rainbow AJ (2009) Differential effects of hypoxia and acidosis on p53 expression, repair of UVC-damaged DNA and viability after UVC in normal and tumor-derived human cells. DNA Repair 8(3):370–382CrossRefPubMedGoogle Scholar
  13. El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B (1993) WAF1, a potential mediator of p53 tumor suppression. Cell 75:817–825CrossRefPubMedGoogle Scholar
  14. Hall P, McKee P, Menage HD, Dover R, Lane DP (1993) High levels of p53 protein in UV-irradiated normal human skin. Oncogene 8:203–207PubMedGoogle Scholar
  15. Hermeking H, Lengauer C, Polyak K (1997) 14-3-3 sigma is a p53 regulated inhibitor of G2/M progression. Mol Cell 1:3–11CrossRefPubMedGoogle Scholar
  16. Jeong SJ, Han SH, Kim DY, Lee JC, Kim HS, Kim BH, Lee JS, Hwang EH, Park JK (2007) Effects of mRg2, a mixture of ginsenosides containing 60% Rg2, on the ultraviolet B-induced DNA repair synthesis and apoptosis in NIH3T3 cells. Int J Toxicol 26(2):151–158CrossRefPubMedGoogle Scholar
  17. Kastan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW (1991) Participation of p53 protein in the cellular response to DNA damage. Cancer Res 51:6304–6311PubMedGoogle Scholar
  18. Kastan M, Zhan Q, el-Deiry W, Carrier F, Jacks T, Walsh WP, Plunkett B, Vogelstein B, Fornace A (1992) A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia–telangiectasia. Cell 71:587–597CrossRefPubMedGoogle Scholar
  19. Kim JH, Cho SY, Lee JH, Jeong SM, Yoon IS, Lee BH, Pyo MK, Lee SM, Chung JM, Kim S, Rhim H, Oh JW, Nah SY (2007) Neuroprotective effects of ginsenoside Rg3 against homocysteine-induced excitotoxicity in rat hippocampus. Brain Res 1136:190–199CrossRefPubMedGoogle Scholar
  20. Kim YK, Yoo DS, Xu H, Park NI, Kim HH, Choi JE, Park SU (2009) Ginsenoside content of berries and roots of three typical Korean ginseng (Panax ginseng) cultivars. Nat Prod Commun 4(7):903–906PubMedGoogle Scholar
  21. Kunwar A, Bansal P, Kumar SJ, Bag PP, Paul P, Reddy ND, Kumbhare LB, Jain VK, Chaubey RC, Unnikrishnan MK, Priyadarsini KI (2010) In vivo radioprotection studies of 3,3′-diselenodipropionic acid, a selenocystine derivative. Free Radical Biol Med 48:399–410CrossRefGoogle Scholar
  22. Laethem AV, Claerhout S, Garmyn M, Agostinis P (2005) The sunburn cell: regulation of death and survival of the keratinocyte. Int J Biochem Cell Biol 37:1547–1553CrossRefPubMedGoogle Scholar
  23. Laethem AV, Garmyn M, Agostinis P (2009) Starting and propagating apoptotic signals in UVB irradiated keratinocytes. Photochem Photobiol Sci 8(3):299–308CrossRefPubMedGoogle Scholar
  24. Lee YJ, Chung E, Lee KY, Lee YH, Huh B, Lee SK (1997) Ginsenoside-Rg1, one of the major active molecules from Panax ginseng, is a functional ligand of glucocorticoid receptor. Mol Cell Endocrinol 133:135–140CrossRefPubMedGoogle Scholar
  25. Lee EH, Cho SY, Kim SJ, Shin ES, Chang HK, Kim DH, Yeom MH, Woe KS, Lee J, Sim YC, Lee TR (2003) Ginsenoside F1 protects human HaCaT keratinocytes from ultraviolet-B-induced apoptosis by maintaining constant levels of Bcl-2. J Invest Dermatol 121:607–613CrossRefPubMedGoogle Scholar
  26. Lee HU, Bae EA, Han MJ, Kim DH (2005) Hepatoprotective effect of ginsenoside Rb1 and compound K on tert-butyl hydroperoxide-induced liver injury. Liver Int 25:1069–1073CrossRefPubMedGoogle Scholar
  27. Liang Y, Lin SY, Brunicardi FC, Goss J, Li K (2008) DNA damage response pathways in tumor suppression and cancer treatment. World J Surg 33(4):661–666CrossRefGoogle Scholar
  28. Meng L, Lin T, Tsai RY (2008) Nucleoplasmic mobilization of nucleostemin stabilizes MDM2 and promotes G2-M progression and cell survival. J Cell Sci 121:4037–4046CrossRefPubMedGoogle Scholar
  29. Mouret S, Charveron M, Favier A, Cadet J, Douki T (2008) Differential repair of UVB-induced cyclobutane pyrimidine dimers in cultured human skin cells and whole human skin. DNA Repair 7(5):704–712CrossRefPubMedGoogle Scholar
  30. Nah SY, Kim DH, Rhim H (2007) Ginsenosides: are any of them candidates for drugs acting on the central nervous system? CNS Drug Rev 13:381–404PubMedGoogle Scholar
  31. Nah SY, Bhatia KS, Lyles J, Ellinwood EH, Lee TH (2008) Effects of ginseng saponin on acute cocaine-induced alterations in evoked dopamine release and uptake in rat brain nucleus accumbens. Brain Res 1248:184–190CrossRefPubMedGoogle Scholar
  32. Powley IR, Kondrashov A, Young LA, Dobbyn HC, Hill K, Cannell IG, Stoneley M, Kong YW, Cotes JA, Smith GC, Wek R, Hayes C, Gant TW, Spriggs KA, Bushell M, Willis AE (2009) Translational reprogramming following UVB irradiation is mediated by DNA–PKcs and allows selective recruitment to the polysomes of mRNAs encoding DNA repair enzymes. Genes Dev 23(10):1207–1220CrossRefPubMedGoogle Scholar
  33. Rakhorst HA, Tra WM, Posthumus-Van Sluijs ST, Hovius SE, Levendag PC, Kanaar R, Hofer SO (2006) Quantitative analysis of radiation-induced DNA break repair in a cultured oral mucosal model. Tissue Eng 12:3395–3403CrossRefPubMedGoogle Scholar
  34. Sarma SN, Kim YJ, Ryu JC (2010) Gene expression profiles of human promyelocytic leukemia cell lines exposed to volatile organic compounds. Toxicol 271(3):122–130Google Scholar
  35. Selivanova G (2010) Therapeutic targeting of p53 by small molecules. Semin Cancer Biol 20:46–56CrossRefPubMedGoogle Scholar
  36. Shieh SY, Ikeda M, Taya Y, Prives C (1997) DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2. Cell 91:325–334CrossRefPubMedGoogle Scholar
  37. Shin YW, Bae EA, Kim SS, Lee YC, Kim DH (2005) Effect of ginsenoside Rb1 and compound K in chronic oxazolone-induced mouse dermatitis. Int Immunopharmacol 5:1183–1191CrossRefPubMedGoogle Scholar
  38. Shivakumar C, Brown D, Deb S (1995) Wild type human p53 transactivates the human proliferating cell nuclear antigen promoter. Mol Cell Biol 15:6785–6793PubMedGoogle Scholar
  39. Siliciano JD, Canman CE, Taya Y, Sakaguchi K, Appella E, Kastan MB (1997) DNA damage induces phosphorylation of the amino terminus of p53. Genes Dev 11:3471–3481CrossRefPubMedGoogle Scholar
  40. Steinberg ML, Hubbard K, Utti C, Clas B, Hwang BJ, Hill HZ, Orlow I (2009) Patterns of persistent DNA damage associated with sun exposure and the glutathione S-transferase M1 genotype in melanoma patients. Photochem Photobiol 85(1):379–386CrossRefPubMedGoogle Scholar
  41. Tsai YS, Lee KW, Huang JL, Liu YS, Juo SH, Kuo WR, Chang JG, Lin CS, Jong YJ (2008) Arecoline, a major alkaloid of areca nut, inhibits p53, represses DNA repair and triggers DNA damage response in human epithelial cells. Toxicology 249(2–3):230–237CrossRefPubMedGoogle Scholar
  42. Utrera R, Collavin L, Lazarevic D, Delia D, Schneider C (1998) A novel p53-inducible gene coding for a microtubule-localized protein with G2-phase specific expression. EMBO J 17:5015–5025CrossRefPubMedGoogle Scholar
  43. Wang J, Wu L, Zhang W, Deng C (2009) Effect of Panax notoginseng saponins on vascular intima hyperplasia and PCNA expression in rat aorta after balloon angioplasty. Zhongguo Zhong Yao Za Zhi 34(6):735–739PubMedGoogle Scholar
  44. Yun TK, Choi SY (1990) A case–control study of ginseng intake and cancer. Int J Epidermiol 19:871–876CrossRefGoogle Scholar
  45. Yun TK, Choi SY (1995) Preventive effect of ginseng intake against various human cancer: a case–control study on 1987 pairs. Cancer Epidemiol Biomarker Prev 4:401–408Google Scholar
  46. Zhan Q, Bae I, Kastan MB, Fornace AJ (1994) Interaction of the p53-regulated protein gadd45 with proliferating cell nuclear antigen. Cancer Res 54:2755–2760PubMedGoogle Scholar
  47. Zhou BR, Luo D, Wei FD, Chen XE, Gao J (2008) Baicalin protects human fibroblasts against ultraviolet B-induced cyclobutane pyrimidine dimers formation. Arch Dermatol Res 300(6):331–334CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Se Eun Ha
    • 1
  • Dae Hyun Shin
    • 2
  • Hyung Do Kim
    • 1
  • Sun Mi Shim
    • 1
  • Hack Soo Kim
    • 2
  • Bo Hyeon Kim
    • 2
  • Jung Sup Lee
    • 3
  • Jong Kun Park
    • 1
    Email author
  1. 1.Division of Biological Science, Institute of Natural SciencesWonkwang UniversityChonbukSouth Korea
  2. 2.Somang Cosmetics R&D CenterIncheonSouth Korea
  3. 3.Department of BiotechnologyChosun UniversityKwangjuSouth Korea

Personalised recommendations