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Inhibitory Effect of Ginseng on Breast Cancer Cell Line Growth Via Up-Regulation of Cyclin Dependent Kinase Inhibitor, p21 and p53

  • Shabanah, Othman A AL (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University) ;
  • Alotaibi, Moureq R (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University) ;
  • Rejaie, Salim S Al (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University) ;
  • Alhoshani, Ali R (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University) ;
  • Almutairi, Mashal M (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University) ;
  • Alshammari, Musaad A (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University) ;
  • Hafez, Mohamed M (College of Pharmacy, Pharmacology and Toxicology Department, Kind Saud University)
  • Published : 2016.11.01

Abstract

Objective: Breast cancer is global female health problem worldwide. Most of the currently used agents for breast cancer treatment have toxic side-effects. Ginseng root, an oriental medicine, has many health benefits and may exhibit direct anti-cancer properties. This study was performed to assess the effects of ginseng on breast cancer cell lines. Materials and Methods: Cytotoxicity of ginseng extract was measured by MTT assay after exposure of MDA-MB-231, MCF-10A and MCF-7 breast cancer cells to concentrations of 0.25, 0.5, 1, 1.5, 2 and 2.5 mg/well. Expression levels of p21WAF, p16INK4A, Bcl-2, Bax and P53 genes were analyzed by quantitative real time PCR. Results: The treatment resulted in inhibition of cell proliferation in a dose-and time-dependent manner. p53, p21WAF1and p16INK4A expression levels were up-regulated in ginseng treated MDA-MB-231 and MCF-7 cancer cells compared to untreated controls and in MCF-10A cells. The expression levels of Bcl2 in the MDA-MB-231 and MCF-7 cells were down-regulated. In contrast, that of Bax was significantly up-regulated. Conclusion: The results of this study revealed that ginseng may inhibit breast cancer cell growth by activation of the apoptotic pathway.

Keywords

Ginseng;MDA-231;MTT assay;gene expression;quantitative real time PCR

References

  1. Al-Ansari MM, Hendrayani SF, Tulbah A, et al (2012). p16INK4A represses breast stromal fibroblasts migration/invasion and their VEGF-A-dependent promotion of angiogenesis through Akt inhibition. Neoplasia, 14, 1269-77. https://doi.org/10.1593/neo.121632
  2. Al-Khalaf HH, Aboussekhra A (2013). p16(INK4A) positively regulates p21(WAF1) expression by suppressing AUF1-dependent mRNA decay. PLoS One, 8, e70133. https://doi.org/10.1371/journal.pone.0070133
  3. Al-Mohanna MA, Al-Khalaf HH, Al-Yousef N, et al (2007). The p16INK4a tumor suppressor controls p21WAF1 induction in response to ultraviolet light. Nucleic Acids Res, 35, 223-33.
  4. Al Dhaheri Y, Eid A, AbuQamar S, et al (2013). Mitotic arrest and apoptosis in breast cancer cells induced by Origanum majorana extract: upregulation of TNF-alpha and downregulation of survivin and mutant p53. PLoS One, 8, e56649. https://doi.org/10.1371/journal.pone.0056649
  5. Azevedo FV, Lopes DS, Gimenes SN, et al (2015). Human breast cancer cell death induced by BnSP-6, a Lys-49 PLA homologue from Bothrops pauloensis venom. Int J Biol Macromol, 82, 671-7.
  6. Bauer KR, Brown M, Cress RD, et al (2007). Descriptive analysis of estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-negative invasive breast cancer, the so-called triple-negative phenotype: a population-based study from the California cancer Registry. Cancer, 109, 1721-8. https://doi.org/10.1002/cncr.22618
  7. Brambilla E, Negoescu A, Gazzeri S, et al (1996). Apoptosisrelated factors p53, Bcl2, and Bax in neuroendocrine lung tumors. Am J Pathol, 149, 1941-52.
  8. Brenton JD, Carey LA, Ahmed AA, et al (2005). Molecular classification and molecular forecasting of breast cancer: ready for clinical application?. J Clin Oncol, 23, 7350-60. https://doi.org/10.1200/JCO.2005.03.3845
  9. Chang YS, Seo EK, Gyllenhaal C, et al (2003). Panax ginseng: a role in cancer therapy?. Integr Cancer Ther, 2, 13-33. https://doi.org/10.1177/1534735403251167
  10. Chiarugi V, Magnelli L, Ruggiero M (1994). Apoptosis, senescence, immortalization and cancer. Pharmacol Res, 30, 301-15. https://doi.org/10.1016/1043-6618(94)80010-3
  11. Chomczynski P (1993). A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques, 15, 532-4.
  12. Coleman KG, Wautlet BS, Morrissey D, et al (1997). Identification of CDK4 sequences involved in cyclin D1 and p16 binding. J Biol Chem, 272, 18869-74. https://doi.org/10.1074/jbc.272.30.18869
  13. Demain AL, Vaishnav P (2011). Natural products for cancer chemotherapy. Microb Biotechnol, 4, 687-99. https://doi.org/10.1111/j.1751-7915.2010.00221.x
  14. Duda RB, Kang SS, Archer SY, et al (2001). American ginseng transcriptionally activates p21 mRNA in breast cancer cell lines. J Korean Med Sci, 16, 54-60. https://doi.org/10.3346/jkms.2001.16.S.S54
  15. el-Deiry WS, Harper JW, O'Connor PM, et al (1994). WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res, 54, 1169-74.
  16. el-Deiry WS, Tokino T, Velculescu VE, et al (1993). WAF1, a potential mediator of p53 tumor suppression. Cell, 75, 817-25. https://doi.org/10.1016/0092-8674(93)90500-P
  17. Farid P, Gomb SZ, Peter I, et al (2001). bcl2, p53 and bax in thyroid tumors and their relation to apoptosis. Neoplasma, 48, 299-301.
  18. Ferguson LR, Chen H, Collins AR, et al (2015). Genomic instability in human cancer: Molecular insights and opportunities for therapeutic attack and prevention through diet and nutrition. Semin Cancer Biol, 35, 5-24. https://doi.org/10.1016/j.semcancer.2015.03.005
  19. Gai X, Tu K, Li C, et al (2015). Histone acetyltransferase PCAF accelerates apoptosis by repressing a GLI1/BCL2/BAX axis in hepatocellular carcinoma. Cell Death Dis, 6, e1712. https://doi.org/10.1038/cddis.2015.76
  20. Gonzalez CA, Riboli E (2010). Diet and cancer prevention: Contributions from the European Prospective Investigation into Cancer and Nutrition (EPIC) study. Eur J Cancer, 46, 2555-62. https://doi.org/10.1016/j.ejca.2010.07.025
  21. Grana X, Reddy EP (1995). Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene, 11, 211-9.
  22. Hassan ZK, Elamin MH, Omer SA, et al (2014). Oleuropein induces apoptosis via the p53 pathway in breast cancer cells. Asian Pac J Cancer Prev, 14, 6739-42.
  23. He BC, Gao JL, Luo X, et al (2011). Ginsenoside Rg3 inhibits colorectal tumor growth through the down-regulation of Wnt/ss-catenin signaling. Int J Oncol, 38, 437-45.
  24. Herrmann M, Lorenz HM, Voll R, et al (1994). A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res, 22, 5506-7. https://doi.org/10.1093/nar/22.24.5506
  25. Jang HJ, Han IH, Kim YJ, et al (2014). Anticarcinogenic effects of products of heat-processed ginsenoside Re, a major constituent of ginseng berry, on human gastric cancer cells. J Agric Food Chem, 62, 2830-6. https://doi.org/10.1021/jf5000776
  26. Kim HS, Lee EH, Ko SR, et al (2004). Effects of ginsenosides Rg3 and Rh2 on the proliferation of prostate cancer cells. Arch Pharm Res, 27, 429-35. https://doi.org/10.1007/BF02980085
  27. Kim SJ, Kim AK (2015). Anti-breast cancer activity of Fine Black ginseng (Panax ginseng Meyer) and ginsenoside Rg5. J Ginseng Res, 39, 125-34. https://doi.org/10.1016/j.jgr.2014.09.003
  28. King ML, Murphy LL (2010). Role of cyclin inhibitor protein p21 in the inhibition of HCT116 human colon cancer cell proliferation by American ginseng (Panax quinquefolius) and its constituents. Phytomedicine, 17, 261-8. https://doi.org/10.1016/j.phymed.2009.06.008
  29. Kralj M, Husnjak K, Korbler T, et al (2003). Endogenous p21WAF1/CIP1 status predicts the response of human tumor cells to wild-type p53 and p21WAF1/CIP1 overexpression. Cancer Gene Ther, 10, 457-67. https://doi.org/10.1038/sj.cgt.7700588
  30. Kumar H, Savaliya M, Biswas S, et al (2015). Assessment of the in vitro cytotoxicity and in vivo anti-tumor activity of the alcoholic stem bark extract/fractions of Mimusops elengi Linn. Cytotechnology, 68, 861-77 .
  31. Kurata K, Yanagisawa R, Ohira M, et al (2008). Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregulation in doxorubicin-treated neuroblastoma cells. Oncogene, 27, 741-54. https://doi.org/10.1038/sj.onc.1210672
  32. Lee DG, Jang SI, Kim YR, et al (2014). Anti-proliferative effects of ginsenosides extracted from mountain ginseng on lung cancer. Chin J Integr Med, 22, 344-52.
  33. Lee JJ, Kwon HK, Jung IH, et al (2009). Anti-cancer activities of ginseng extract fermented with phellinus linteus. Mycobiology, 37, 21-7. https://doi.org/10.4489/MYCO.2009.37.1.021
  34. Li N, Li Q, Cao X, et al (2011). The tumor suppressor p33ING1b upregulates p16INK4a expression and induces cellular senescence. FEBS Lett, 585, 3106-12. https://doi.org/10.1016/j.febslet.2011.08.044
  35. Li Y, Upadhyay S, Bhuiyan M, et al (1999). Induction of apoptosis in breast cancer cells MDA-MB-231 by genistein. Oncogene, 18, 3166-72. https://doi.org/10.1038/sj.onc.1202650
  36. Liu W, Zhang R (1998). Upregulation of p21WAF1/CIP1 in human breast cancer cell lines MCF-7 and MDA-MB-468 undergoing apoptosis induced by natural product anticancer drugs 10-hydroxycamptothecin and camptothecin through p53-dependent and independent pathways. Int J Oncol, 12, 793-804.
  37. Mann J (2002). Natural products in cancer chemotherapy: past, present and future. Nat Rev Cancer, 2, 143-8. https://doi.org/10.1038/nrc723
  38. Munagala R, Kausar H, Munjal C, et al (2011). Withaferin A induces p53-dependent apoptosis by repression of HPV oncogenes and upregulation of tumor suppressor proteins in human cervical cancer cells. Carcinogenesis, 32, 1697-705. https://doi.org/10.1093/carcin/bgr192
  39. Naseri MH, Mahdavi M, Davoodi J, et al (2015). Up regulation of Bax and down regulation of Bcl2 during 3-NC mediated apoptosis in human cancer cells. Cancer Cell Int, 15, 55. https://doi.org/10.1186/s12935-015-0204-2
  40. Newman DJ, Cragg GM, Holbeck S, et al (2002). Natural products and derivatives as leads to cell cycle pathway targets in cancer chemotherapy. Curr Cancer Drug Targets, 2, 279-308. https://doi.org/10.2174/1568009023333791
  41. Nishino H, Tokuda H, Ii T, et al (2001). Cancer chemoprevention by ginseng in mouse liver and other organs. J Korean Med Sci, 16, 66-9. https://doi.org/10.3346/jkms.2001.16.S.S66
  42. Nurse P, Masui Y, Hartwell L (1998). Understanding the cell cycle. Nat Med, 4, 1103-6. https://doi.org/10.1038/2594
  43. Pal SK, Childs BH, Pegram M (2011). Triple negative breast cancer: unmet medical needs. Breast Cancer Res Treat, 125, 627-36. https://doi.org/10.1007/s10549-010-1293-1
  44. Phalke S, Mzoughi S, Bezzi M, et al (2012). p53-Independent regulation of p21Waf1/Cip1 expression and senescence by PRMT6. Nucleic Acids Res, 40, 9534-42. https://doi.org/10.1093/nar/gks858
  45. Polyak K, Xia Y, Zweier JL, et al (1997). A model for p53-induced apoptosis. Nature, 389, 300-5. https://doi.org/10.1038/38525
  46. Prasad VS, LaFond RE, Zhou M, et al (1997). Upregulation of endogenous p53 and induction of in vivo apoptosis in B-lineage lymphomas of E(mu)-myc transgenic mice by deregulated c-myc transgene. Mol Carcinog, 18, 66-77. https://doi.org/10.1002/(SICI)1098-2744(199702)18:2<66::AID-MC2>3.0.CO;2-O
  47. Raina K, Agarwal R (2007). Combinatorial strategies for cancer eradication by silibinin and cytotoxic agents: efficacy and mechanisms. Acta Pharmacol Sin, 28, 1466-75. https://doi.org/10.1111/j.1745-7254.2007.00691.x
  48. Santana-Davila R, Perez EA (2010). Treatment options for patients with triple-negative breast cancer. J Hematol Oncol, 3, 42. https://doi.org/10.1186/1756-8722-3-42
  49. Saw CL, Wu Q, Kong AN (2010). Anti-cancer and potential chemopreventive actions of ginseng by activating Nrf2 (NFE2L2) anti-oxidative stress/anti-inflammatory pathways. Chin Med, 5, 37. https://doi.org/10.1186/1749-8546-5-37
  50. Sharma J, Goyal PK (2015). Chemoprevention of chemicalinduced skin cancer by Panax ginseng root extract. J Ginseng Res, 39, 265-73. https://doi.org/10.1016/j.jgr.2015.01.005
  51. Szallasi Z, Liang S (1998). Modeling the normal and neoplastic cell cycle with "realistic Boolean genetic networks": their application for understanding carcinogenesis and assessing therapeutic strategies. Pac Symp Biocomput, 66-76.
  52. Teijido O, Dejean L (2010). Upregulation of Bcl2 inhibits apoptosis-driven BAX insertion but favors BAX relocalization in mitochondria. FEBS Lett, 584, 3305-10. https://doi.org/10.1016/j.febslet.2010.07.002
  53. Thees S, Hubbard GB, Winckler J, et al (2005). Specific alteration of the Bax/Bcl2 ratio and cytochrome c without execution of apoptosis in the hippocampus of aged baboons. Restor Neurol Neurosci, 23, 1-9.
  54. Torre LA, Bray F, Siegel RL, et al (2015). Global cancer statistics, 2012. CA Cancer J Clin, 65, 87-108. https://doi.org/10.3322/caac.21262
  55. Vibet S, Goupille C, Bougnoux P, et al (2008). Sensitization by docosahexaenoic acid (DHA) of breast cancer cells to anthracyclines through loss of glutathione peroxidase (GPx1) response. Free Radic Biol Med, 44, 1483-91. https://doi.org/10.1016/j.freeradbiomed.2008.01.009
  56. Wang X, Wei Y, Yuan S, et al (2006). Potential anticancer activity of litchi fruit pericarp extract against hepatocellular carcinoma in vitro and in vivo. Cancer Lett, 239, 144-50. https://doi.org/10.1016/j.canlet.2005.08.011
  57. Wang X, Zheng YL, Li K, et al (2009). The effects of ginsenosides Rg3 on the expressions of VEGF and KDR in human lung squamous cancer cells. Zhong Yao Cai, 32, 1708-10.
  58. Wong AS, Che CM, Leung KW (2015). Recent advances in ginseng as cancer therapeutics: a functional and mechanistic overview. Nat Prod Rep, 32, 256-72. https://doi.org/10.1039/C4NP00080C
  59. Wu X, Zahari MS, Ma B, et al (2015). Global phosphotyrosine survey in triple-negative breast cancer reveals activation of multiple tyrosine kinase signaling pathways. Oncotarget, 6, 29143-60. https://doi.org/10.18632/oncotarget.5020
  60. Zheng Y, Nan H, Hao M, et al (2013). Antiproliferative effects of protopanaxadiol ginsenosides on human colorectal cancer cells. Biomed Rep, 1, 555-8. https://doi.org/10.3892/br.2013.104