DOI QR코드

DOI QR Code

Effects of polysaccharides derived from Orostachys japonicus on induction of cell cycle arrest and apoptotic cell death in human colon cancer cells

  • Ryu, Deok-Seon ;
  • Baek, Geum-Ok ;
  • Kim, Eun-Young ;
  • Kim, Ki-Hoon ;
  • Lee, Dong-Seok
  • Received : 2010.08.16
  • Accepted : 2010.09.30
  • Published : 2010.11.30

Abstract

Crude Orostachys japonicus polysaccharide extract (OJP) was prepared by hot steam extraction. Polysaccharides (OJPI) were separated from OJP by gel filtration chromatography and phenol-sulfuric acid assay. The average molecular weight of the OJPI was 30-50 kDa. The anti-proliferative effect of OJPI on HT-29 human colon cancer cells was investigated via morphology study, cell viability assay, apoptosis assay, cell cycle analysis, and cDNA microarray. OJPI inhibited proliferation and growth of HT29 cells and also stimulated apoptosis in a dose- and time-dependent manner. In cell cycle analysis, treatment with OJPI resulted in a marked increase of cells in the G0 (sub G1) and G2/M phases. To screen for genes involved in the induction of cell cycle arrest and apoptosis, the gene expression profiles of HT-29 cells treated with OJPI were examined by cDNA microarray, revealing that a number of genes were up- or down-regulated by OJPI. Whereas several genes involved in anti-apoptosis, cell proliferation and growth, and cell cycle regulation were down-regulated, expression levels of several genes involved in apoptosis, tumor suppression, and other signal transduction events were up-regulated. These results suggest that OJPI inhibits the growth of HT-29 human colon cancer cells by various apoptosis-aiding activities as well as apoptosis itself. Therefore, OJPI deserve further development as an effective agent exhibiting anticancer activity.

Keywords

Anti-proliferative activity;Apoptosis;Cell cycle arrest;Microarray;Orostachys japonicus;Polysaccharides

References

  1. Je, M., Jung, W. J., Lee, K. Y., Kim, Y. C. and Sung, S. H. (2009) Calpain inhibitory flavonoids isolated from Orostachys japonicus. J. Enzyme Inhib. Med. Chem. 24, 676-679. https://doi.org/10.1080/14756360802328075
  2. Park, H. J., Han, S. Y., Kun, Y. P., Sook, H. R., Hae, Y. C. and Jae, S. C. (1991) Flavonoids from the whole plants of Orostachys japonicus. Arch. Pharm. Res. 14, 167-171. https://doi.org/10.1007/BF02892023
  3. Yoon, Y., Kim, K. S., Hong, S. G., Kang, B. J., Lee, M. Y. and Cho, D. W. (2000) Protective effects of Orostachys japonicus A. Berger (Crossuloceae) on $H_2O_2$ - induced apoptosis in GTI-1 mouse hypothalamic neuronal cell line. J. Ethnopharmacol. 69, 73-78. https://doi.org/10.1016/S0378-8741(99)00107-5
  4. Yoon, N. Y., Min, B. S., Lee, H. K., Park, J. C. and Choi, J. S. (2005) A potent anti - complementary acylated sterol glucoside from Orostachys japonicus. Arch. Pharm. Res. 28, 892-896. https://doi.org/10.1007/BF02973873
  5. Jemal, A., Siegel, R., Wand, E., Hao, Y., Xu, J. and Thun, M. J. (2009) Cancer statistics. Cancer J. Clin. 59, 225-249. https://doi.org/10.3322/caac.20006
  6. Chau, I. and Cunningham, D. (2006) Adjuvant therapy in colon cancer - What, when and how? Ann. Oncol. 17, 1347-1359. https://doi.org/10.1093/annonc/mdl029
  7. Kelloff, G. J. (2000) Perspectives on cancer chemoprevention research and drug development. Adv. Cancer Res. 78, 199-334.
  8. Ryu, D. S., Kim, S. H. and Lee, D. S. (2009) Anti-proliferative effect of polysaccharides from Salicornia herbacea on induction of G2/M arrest and apoptosis in human colon cancer cells. J. Microbiol. Biotechnol. 19, 1482-1489. https://doi.org/10.4014/jmb.0902.0063
  9. Kaina, B. (2003) DNA damage - triggered apoptosis: critical role of DNA repair, double - strand breaks, cell proliferation and signaling. Biochem. Pharmacol. 66, 1547-1554. https://doi.org/10.1016/S0006-2952(03)00510-0
  10. Kim, K. H., Kim, E. Y., Kim, Y. O., Beak, G. O., Kim, H. B. and Lee, D. S. (2004) Studies on biological activities of the polysaccharides and oligosaccharides of Orostachys Japonicus. Korean J. Microbiol. 40, 334-341.
  11. Nam, K. S., Kim, M. K. and Shon, Y. H. (2007) Chemopreventive effect of chitosan oligosaccharide against colon carcinogenesis. J. Microbiol. Biotechnol. 17, 1546-1549.
  12. Hyun, J. H., Kim, S. C., Kang, J. I., Kim, M. K., Boo, H. J., Kwon, J. M., Koh, Y. S., Hyun, J. W., Park, D. B., Yoo, E. S. and Kang, H. K. (2009) Apoptosis inducing activity of fucoidan in HCT-15 colon carcinoma cells. Biol. Pharm. Bull. 32, 1760-1764. https://doi.org/10.1248/bpb.32.1760
  13. Bao, H. H., Choi, W. S. and You, S. G. (2010) Effect of sulfated modification on the molecular characteristics and biological activities of polysaccharides from Hypsizigus marmoreus. Biosci. Biotechnol. Biochem. 74, 1408-1414. https://doi.org/10.1271/bbb.100076
  14. Arends, M. J. (1999) Apoptosis in the endometrium. Mistopathology. 35, 174-178. https://doi.org/10.1046/j.1365-2559.1999.00756.x
  15. Boukes, G. J., Daniels, B. B., Albrecht, C. F. and Van de venter, M. (2010) Cell survival or apoptosis: rooperol’s role as anticancer agent. Oncol. Res. 18, 365-376. https://doi.org/10.3727/096504010X12644422320627
  16. Soule, B. P., Simone, N. L., DeGraff, W. G., Choudhuri, R., Cook, J. A. and Mitchell, J. B. (2010) Loratadine dysregulates cell cycle progression and enhances the effect of radiation in human tumor cell lines. Radiat. Oncol. 5, 1-12. https://doi.org/10.1186/1748-717X-5-1
  17. Yang, E., Zha, J., Jockel, J., Boise, L. H. and Thompson, C. B. (1995) Bad, a heterodimeric partner for Bcl-XL and Bcl-2, displaces Bax and promotes cell death. Cell 80, 285-291. https://doi.org/10.1016/0092-8674(95)90411-5
  18. Zhang, J. K., Cado, D., Chen, A., Kabra, N. H. and Winoto, A. (1998) Fas - mediated apoptosis and activation - induced T-cell proliferation are defective in mice lacking FADD/Mort 1. Nature 392, 296-300. https://doi.org/10.1038/32681
  19. Liu, C., Wu, X., Luo, C., Hu, Z., Yin, Z., He, Y., Du, H., Zhang, W., Jiang, Q. and Lin, Y. (2010) Antisense oligonucleotide targeting Livin induces apoptosis of human bladder cancer cell via a mechanism involving caspase3. J. Exp. Clin. Cancer Res. 29, 1-8. https://doi.org/10.1186/1756-9966-29-1
  20. Naumovski, L. and Cleary, M. L. (1996) The p53 - binding protein 53BP2 also interacts with Bc1-2 and impedes cell cycle progression at G2/M. Mol. Cell. Biol. 16, 3884-3892. https://doi.org/10.1128/MCB.16.7.3884
  21. Kimura, H. J., Rocchi, R., Landek-Salgado, M. A., Suzuki, K., Chen, C. Y., Kimura, M., Rose, N. R. and Caturegli, P. (2009) Influence of signal transducer and activator of transcription-1 Signaling on thyroid morphology and function. Endocrinology. 150, 3409-3416. https://doi.org/10.1210/en.2008-1769
  22. Lee, S. T., Strunk, K. M. and Spritz, R. A. (1993) A survey of protein tyrosin kinase mRNAs expressed in normal human melanocytes. Oncogene. 8, 3403-3410.
  23. Gibson, L., Holmgreen, S. P., Huang, D. C., Bernard, O., Copeland, N. G., Jenkins, N. A., Sutherland, G. R., Baker, E., Adams, J. M. and Cory, S. (1996) Bcl-w, a novel member of the bcl-2 family, promotes cell survival. Oncogene. 15, 665-675.
  24. Naiki, T., Shimomura, T., Kondo, T., Matsumoto, K. and Sugimoto, K. (2000) Rfc5, in cooperation with rad 24, controls DNA damage checkpoints throughout the cell cycle in Saccharomyces cerevisiae. Mol. cell Biol. 20, 5888-5896. https://doi.org/10.1128/MCB.20.16.5888-5896.2000
  25. Buhling, F., Waldburg, N., Kruger, S., Rocken, C., Wiesner, O., Weber, E. and Welte, T. (2002) Expression of cathepsins B, H, K, L, and S during human fetal lung development. Dev. Dyn. 225, 14-21. https://doi.org/10.1002/dvdy.10134
  26. Klarzynski, O., Plesse, J. M., Joubert, J. M., Yvin, J. C., Kopp, M., Kloareg, B. and Fritig, B. (2000) Linear beta - 1, 3 glucans are elicitors of defense responses in tobacco. Plant Physiol. 124, 1027-1038. https://doi.org/10.1104/pp.124.3.1027
  27. Cuesta, G., Suarez, N., Bessio, M. I., Ferreira, F. and Massaldi, H. (2003) Quantitative determination of pneumococcal capsular polysaccharide serotype 14 using a modification of phenol - sulfuric acid method. J. Microbiol. Methods. 52, 69-73. https://doi.org/10.1016/S0167-7012(02)00151-3
  28. Eray, M., Matto, M., Kaartinen, M., Andersson, L. and Pelkonen, J. (2001) Flow cytometric analysis of apoptotic subpopulations with a combination of annexin V-FITC, propidium iodide, and SYTO 17. Cytometry. 43, 134-142. https://doi.org/10.1002/1097-0320(20010201)43:2<134::AID-CYTO1028>3.0.CO;2-L
  29. Kim, K. H., Kim, Y. W., Kim, H. B., Lee, B. J. and Lee, D. S. (2006) Anti - apoptotic activity of laminarin polysaccharides and their enzymatically hydrolyzed oligosaccharides from Laminaria Joponica. Biotechnol. Lett. 28, 439-446. https://doi.org/10.1007/s10529-005-6177-9

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Acknowledgement

Supported by : Inje University