Inhibitory effect of selenomethionine on carcinogenesis in the model of human colorectal cancer in vitro and its link to the Wnt/β-catenin pathway.

  • Edyta Korbut Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Krakow, 7 Gronostajowa Str., 30-387 Krakow, Poland
  • Agata Ptak-Belowska Department of Physiology, Jagiellonian University Medical College, 16 Grzegorzecka Str., 31-531 Krakow, Poland
  • Tomasz Brzozowski Department of Physiology, Jagiellonian University Medical College, 16 Grzegorzecka Str., 31-531 Krakow, Poland
Keywords: Colorectal cancer, GSK-3β, Wnt/β-catenin pathway, Selenium, Selenomethionine


Selenium compounds have been implicated as anticancer agents; however, the mechanism of their inhibitory action against cancer development has not been extensively investigated. The constitutive activation of the Wnt/β-catenin pathway is a central event in colorectal carcinogenesis. In this pathway, the excessive cell proliferation is initiated by the generation of β-catenin followed by overexpression of proto-oncogenes such as c-Myc. It is believed that under physiological conditions the level of c-Myc is efficiently controlled by accessibility of β-catenin protein through the process of phosphorylation by glycogen synthase kinase 3β (GSK-3β). Here, we determined whether selenomethionine (SeMet) can inhibit cell growth and affect the Wnt/β-catenin pathway in HT-29 human colorectal cancer cells in vitro. The effective cytotoxic doses of SeMet have been selected after 48 h of incubation of this compound with colorectal cancer HT-29 cell line. The MTT assay was used to assess cell viability and the protein and mRNA levels of β-catenin and c-Myc were determined by Western blotting and qPCR, respectively. The SeMet potently inhibited growth of HT-29 cells, significantly decreased the β-catenin protein and mRNA concentration, down-regulated the c-Myc gene expression and up-regulated pro-apoptotic Bax protein expression. Moreover, SeMet increased the level of GSK-3β phosphorylated at serine 9 (S9) and significantly increased the level of β-catenin phosphorylated at S33 and S37. We conclude that SeMet suppresses the growth of HT-29 colorectal cancer cells by the mechanism linked to the Wnt/β-catenin pathway, however, the degradation of β-catenin may occur independently of GSK-3β catalytic activity and its phosphorylation status.


Adams JM, Cory S (2007) The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26: 1324-1337. doi: 10.1038/sj.onc.1210220

Anastas JN, Moon RT (2013) WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer 13: 11-26. doi: 10.1038/nrc3419

Bandapalli OR, Dihlmann S, Helwa R, Macher-Goeppinger S, Weitz J, Schirmacher P, et al. (2009) Transcriptional activation of the beta-catenin gene at the invasion front of colorectal liver metastases. J Pathol 218: 370-379. doi: 10.1002/path.2539

Bermingham EN, Hesketh JE, Sinclair BR, Koolaard JP, Roy NC (2014) Selenium-enriched foods are more effective at increasing glutathione peroxidase (GPx) activity compared with selenomethionine: a meta-analysis. Nutrients 6: 4002-4031. doi: 10.3390/nu6104002

Beutler E, Beutler B, Matsumoto J (1975) Glutathione peroxidase activity of inorganic selenium and seleno-DL-cysteine. Experientia 31: 769-770.

Bordonaro M, Lazarova DL, Augenlicht LH, Sartorelli AC (2002) Cell type- and promoter-dependent modulation of the Wnt signaling pathway by sodium butyrate. Int J Cancer 97: 42-51.

Brozmanova J, Manikova D, Vlckova V, Chovanec M (2010) Selenium: a double-edged sword for defense and offence in cancer. Arch Toxicol 84: 919-938. doi: 10.1007/s00204-010-0595-8

Carroll L, Davies MJ, Pattison D (2015) Reaction of low-molecular-mass organoselenium compounds (and their sulphur analogues) with inflammation-associated oxidants. Free Radic Res 49: 750-767. doi: 10.3109/10715762.2015.1018247

Clevers H (2006) Wnt/beta-catenin signaling in development and disease. Cell 127: 469-480. doi: 10.1016/j.cell.2006.10.018

Combs GF, Jr., Gray WP (1998) Chemopreventive agents: selenium. Pharmacol Ther 79: 179-192.

Dihlmann S, Klein S, von Knebel Doeberitz M (2003) Reduction of beta-catenin/T-cell transcription factor signaling by aspirin and indomethacin is caused by an increased stabilization of phosphorylated beta-catenin. Mol Cancer Ther. 2: 509-516. doi: 10.1002/ijc.20609

Doble BW (2003) GSK-3: tricks of the trade for a multi-tasking kinase. Journal of Cell Science 116: 1175-1186. doi: 10.1242/jcs.00384

Ebert MP, Fei G, Kahmann S, Muller O, Yu J, Sung JJ, et al. (2002) Increased beta-catenin mRNA levels and mutational alterations of the APC and beta-catenin gene are present in intestinal-type gastric cancer. Carcinogenesis 23: 87-91.

Emanuele S, D'Anneo A, Bellavia G, Vassallo B, Lauricella M, De Blasio A, et al. (2004) Sodium butyrate induces apoptosis in human hepatoma cells by a mitochondria/caspase pathway, associated with degradation of beta-catenin, pRb and Bcl-XL. Eur J Cancer 40: 1441-1452. doi: 10.1016/j.ejca.2004.01.039

Fevr T, Robine S, Louvard D, Huelsken J ( 2007) Wnt/beta-catenin is essential for intestinal homeostasis and maintenance of intestinal stem cells. Molecular and cellular biology 27: 7551-7559. doi: 10.1128/MCB.01034-07

Gao C, Xiao G, Hu J (2014) Regulation of Wnt/beta-catenin signaling by posttranslational modifications. Cell Biosci 4: 13. doi: 10.1186/2045-3701-4-13

Goulet AC, Watts G, Lord JL, Nelson MA (2007) Profiling of selenomethionine responsive genes in colon cancer by microarray analysis. Cancer Biol Ther 6: 494-503.

Han Y, Park J, Park S, Hahm K, Hong S, Kim E (2013) Gastrin promotes intestinal polyposis through cholecystokinin-B receptor-mediated proliferative signaling and fostering tumor microenvironment. J Physiol Pharmacol 64: 429-437.

He X, Semenov M, Tamai K, Zeng X (2004) LDL receptor-related proteins 5 and 6 in Wnt/beta-catenin signaling: arrows point the way. Development 131: 1663-1677. doi: 10.1242/dev.01117

Kikuchi A (2003) Tumor formation by genetic mutations in the components of the Wnt signaling pathway. Cancer Sci 94: 225-229.

Korbut E, Ptak-Belowska A, Brzozowski T (2012) Mechanisms promoting physiological cells progression into tumorigenesis. J Physiol Pharmacol 63: 565-570.

Liu J, Stevens J, Rote CA, Yost HJ, Hu Y, Neufeld KL, et al. (2001) Siah-1 mediates a novel beta-catenin degradation pathway linking p53 to the adenomatous polyposis coli protein. Mol Cell 7: 927-936.

McCubrey JA, Steelman LS, Bertrand FE, Davis NM, Sokolosky M, Abrams SL, et al. (2014) GSK-3 as potential target for therapeutic intervention in cancer. Oncotarget 5: 2881-2911. doi: 10.18632/oncotarget.2037

Menter DG, Sabichi AL, Lippman SM (2000) Selenium effects on prostate cell growth. Cancer Epidemiol Biomarkers Prev 9: 1171-1182.

Miller DM, Thomas SD, Islam A, Muench D, Sedoris K (2012) c-Myc and cancer metabolism. Clin Cancer Res 18: 5546-5553. doi: 10.1158/1078-0432.CCR-12-0977

Mirlashari MR, Randen I, Kjeldsen-Kragh J (2012) Glycogen synthase kinase-3 (GSK-3) inhibition induces apoptosis in leukemic cells through mitochondria-dependent pathway. Leukemia research 36: 499-508. doi: 10.1016/j.leukres.2011.11.013

Narayanan BA, Narayanan NK, Desai D, Pittman B, Reddy BS (2004) Effects of a combination of docosahexaenoic acid and 1,4-phenylene bis(methylene) selenocyanate on cyclooxygenase 2, inducible nitric oxide synthase and beta-catenin pathways in colon cancer cells. Carcinogenesis 25: 2443-2449. doi: 10.1093/carcin/bgh252

Nelson MA, Goulet AC, Jacobs ET, Lance P (2005) Studies into the anticancer effects of selenomethionine against human colon cancer. Ann N Y Acad Sci 1059: 26-32. doi: 10.1196/annals.1339.016

Pecina-Slaus N (2010) Wnt signal transduction pathway and apoptosis: a review. Cancer Cell Int 10: 22. doi: 10.1186/1475-2867-10-22

Polakis P (2012) Wnt signaling in cancer. Cold Spring Harb Perspect Biol 4. doi: 10.1101/cshperspect.a008052

Rao CV, Cooma I, Rodriguez JG, Simi B, El-Bayoumy K, Reddy BS (2000) Chemoprevention of familial adenomatous polyposis development in the APC(min) mouse model by 1,4-phenylene bis(methylene)selenocyanate. Carcinogenesis 21: 617-621.

Rayman MP (2000) The importance of selenium to human health. Lancet 356: 233-241. doi: 10.1016/S0140-6736(00)02490-9

Redman C, Scott JA, Baines AT, Basye JL, Clark LC, Calley C, et al. (1998) Inhibitory effect of selenomethionine on the growth of three selected human tumor cell lines. Cancer Lett 125: 103-110.

Redman C, Xu MJ, Peng YM, Scott JA, Payne C, Clark LC, et al. (1997) Involvement of polyamines in selenomethionine induced apoptosis and mitotic alterations in human tumor cells. Carcinogenesis 18: 1195-1202.

Saifo MS, Rempinski DR, Jr., Rustum YM, Azrak RG (2010) Targeting the oncogenic protein beta-catenin to enhance chemotherapy outcome against solid human cancers. Mol Cancer 9: 310. doi: 10.1186/1476-4598-9-310

Smith ML, Hawcroft G, Hull MA (2000) The effect of non-steroidal anti-inflammatory drugs on human colorectal cancer cells: evidence of different mechanisms of action. Eur J Cancer 36: 664-674.

Tan J, Zhuang L, Leong HS, Iyer NG, Liu ET, Yu Q (2005) Pharmacologic modulation of glycogen synthase kinase-3beta promotes p53-dependent apoptosis through a direct Bax-mediated mitochondrial pathway in colorectal cancer cells. Cancer Res 65: 9012-9020. doi: 10.1158/0008-5472.CAN-05-1226

Zeng H, Wu M, Botnen JH (2009) Methylselenol, a selenium metabolite, induces cell cycle arrest in G1 phase and apoptosis via the extracellular-regulated kinase 1/2 pathway and other cancer signaling genes. J Nutr 139: 1613-1618. doi: 10.3945/jn.109.110320

Zhang W, Yan S, Liu M, Zhang G, Yang S, He S, et al. (2010) beta-Catenin/TCF pathway plays a vital role in selenium induced-growth inhibition and apoptosis in esophageal squamous cell carcinoma (ESCC) cells. Cancer Lett 296: 113-122. doi: 10.1016/j.canlet.2010.04.001

Zhou W, Wang L, Gou SM, Wang TL, Zhang M, Liu T, et al. (2012) ShRNA silencing glycogen synthase kinase-3 beta inhibits tumor growth and angiogenesis in pancreatic cancer. Cancer Lett 316: 178-186. doi: 10.1016/j.canlet.2011.10.033