Skip to main content
Log in

Sodium butyrate induces retinoblastoma protein dephosphorylation, p16 expression and growth arrest of colon cancer cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Sodium butyrate causes alteration of colon cancer cell morphology and biology towards that of a more differentiated phenotype. The retinoblastoma gene encodes a nuclear phosphoprotein (pRb) present in a wide range of human cancer cell lines including colon cancer cell lines. pRB is synthesized throughout the cell cycle and phosphorylated in a phase specific manner: the predominant proteins in G0/G1 are the unphosphorylated species (110 kD) whereas phosphorylated pRb (112-114 kD) are in S and G2. 110 kD pRb binds transcription factors and prevents transcription of responsive genes such as the gene for thymidine kinase, which are expressed in late G1. The precise mechanisms controlling cell arrest are unknown, but recent data suggest that cyclin-dependent kinase inhibitors such as p16 may play a role. The aim of the present study was to assess the effect of sodium butyrate on cell cycle staging, thymidine kinase activity, phosphorylation of the pRb protein and expression of p16. We show that sodium butyrate treatment induces differentiation of LS174T colon cancer cells, inhibits thymidine kinase activity concomitantly with induction of pRb dephosphorylation, p16 transcription and cell cycle arrest at G0/G1. Initial dephosphorylation was observed 24 h after treatment of LS174T cells with sodium butyrate, whereas complete shift to the dephosphorylated form was observed 3 days after treatment. Induction of pRb dephosphorylation by sodium butyrate preceded inhibition of growth and the specific cell cycle arrest. RNase protection assay with a p16 specific riboprobe showed undetectable levels in proliferating cells to several fold increase in differentiated colonocytes.

In conclusion, the results provide evidence for a specific cellular mechanism of butyrate induced growth arrest and differentiation of a colon cancer cell line.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Scheppach W, Bartram HP, Richter F: Role of short-chain fatty acids in the prevention of colorectal cancer. Eur J Cancer 31A: 1077–1080, 1995

    PubMed  Google Scholar 

  2. Hague A, Butt AJ, Paraskeva C: The role of butyrate in human colonic epithelial cells: an energy source or inducer of differentiation and apoptosis? Proc Nut Soc 55: 937–943, 1996

    Google Scholar 

  3. Deng G, Liu G, Hu L, Gum JR, Kim YS: Transcriptional regulation of the human placental-like alkaline phosphatase gene and mechanisms involved in its induction by sodium butyrate. Cancer Res 52: 3378–3383, 1992

    PubMed  Google Scholar 

  4. Kruh J, Tichonicky L, Defer N: Effect of butyrate on gene expression. In: HJ Binder, JH Cummings, K Soergel (eds.) Short Chain Fatty Acids. Dordrecht, Kluwer, pp 135–147, 1994

    Google Scholar 

  5. Gamet L, Daviaud D, Denis-Pouxviel C, Remesy C, Murat J-C: Effect of short-chain fatty acids on growth and differentiation of the human colon cancer cell line HT-29. Int J Cancer 52: 286–289, 1992

    PubMed  Google Scholar 

  6. Gum JR, Kam JC, Hicks JW, Sleisenger MH, Kim YS: Effects of sodium butyrate on human colonic adenocarcinoma cells. Induction of placentallike alkaline phosphatase. J Biol Chem 262: 1092–1097, 1987

    PubMed  Google Scholar 

  7. Wiman KG: The retinoblastoma gene: role in cell cycle control and cell differentiation. Faseb J 7: 841–845, 1993

    PubMed  Google Scholar 

  8. Riley DJ, Lee KY, Lee WH: The retinoblastoma protein: more than a tumor suppresor. Annu Rev Cell Biol 10: 1–29, 1995

    Google Scholar 

  9. Weinberg RA: The retinoblastoma protein and cell cycle control. Cell 81: 323–330, 1995

    Article  PubMed  Google Scholar 

  10. Bartek J, Bartkova J, Lukas J: The retinoblastoma protein pathway and the restriction point. Current Opin Cell Biol 8: 805–814, 1996

    Google Scholar 

  11. Gope R, Gope ML: Abundance and state of phosphorylation of the retinoblastoma susceptibility gene product in human colon cancer. Mol Cell Biochem 110: 123–133, 1992

    PubMed  Google Scholar 

  12. Ali AA, Harvey JP, Wildrick DM, Boman BM: Retinoblastoma gene productassociated proteins in human colon cancer cell lines. Biochem Biophys Res Commun 194: 848–854, 1993

    PubMed  Google Scholar 

  13. Sherr CJ: Mammalian G1 cyclins. Cell 73: 1059–1065, 1993

    Article  PubMed  Google Scholar 

  14. Hunter T, Pines J: Cyclins and cancer II: Cyclin D and CDK inhibitors come of age. Cell 79: 573–582, 1994

    Article  PubMed  Google Scholar 

  15. Nigg EA: Cyclin-dependent protein kinases: Key regulators of the eukaryotic cell cycle. Bio Essays 17: 471–480, 1995

    Google Scholar 

  16. Resnitzky D, Reed SI: Different roles for cyclins D1 and E in regulation of the G1-to-S transition. Mol Cell Biol 15: 3463–3469, 1995

    PubMed  Google Scholar 

  17. Kranenburg O, Van der EB AJ, Zantema A: Cyclin D kinases and pRb: Regulators of the proliferation-differentiation switch. 367: 103–106, 1995

    Google Scholar 

  18. Tanaka EM, Gann AAF, Gates PB, Brockes JP: Newt myotubes reenter the cell cycle by phosphorylation of the retinoblastoma protein. J Cell Biol 136: 155–165, 1997

    PubMed  Google Scholar 

  19. Sherr CJ, Roberts JM: Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev 9: 1149–1163, 1995

    PubMed  Google Scholar 

  20. Harper JW, Elledge SJ: Cdk inhibitors in development and cancer. Curr Opin Genet Dev 6: 56–64, 1996

    PubMed  Google Scholar 

  21. Yeager T, Stadler W, Belair C, Puthenveettil J, Olopade O, Reznikoff C: Increased p16 levels correlate with pRb alterations in human urothelial cells. Cancer Res 55: 493–497, 1995

    PubMed  Google Scholar 

  22. Ragione FD, Russo GL, Oliva A, Mercurio C, Mastropietro S, Pietra VD, Zappia V: Biochemical characterization of p16NK4-and p18-containing complexes in human cell lines. J Biol Chem 271: 15942–15949, 1996

    PubMed  Google Scholar 

  23. Serrano M, Lee HW, Chin L, Cordon-Cardo C, Beach D, DePinho RA: Role of the INK4a locus in tumor suppression and cell mortality. Cell 85: 27–37, 1996

    PubMed  Google Scholar 

  24. Schwartz B, Polak-Charcon S, Lamprecht SA, Niv Y, Kim YS: The induction of a differentiated phenotype in human colon cancer cells requires the attenuation of cytoskeletal tyrosine phosphorylation. Oncol Res 7: 277–287, 1995

    PubMed  Google Scholar 

  25. Schwartz B, Avivi C, Lamprecht SA: Isolation and characterization of normal and neoplastic colonic epithelial cell populations. Gastroenterology 100: 692–702, 1991

    PubMed  Google Scholar 

  26. DeCaprio JA, Ludlow JW, Lynch D, Furukawa Y, Griffin J, Plwnica-Worms H, Huang CM, Livingston DM: The product of the retinoblastoma susceptibility gene has properties of a cell cycle regulatory element. Cell 58: 1085–1095, 1989

    PubMed  Google Scholar 

  27. Lois AF, Cooper LT, Geng Y, Nobori T, Carson D: Expression of the p16 and p15 cyclin-dependent kinase inhibitors in lymphocyte activation and neuronal differentiation. Cancer Res 55: 4010–4013, 1995

    PubMed  Google Scholar 

  28. Pinto M, Appay MD, Simon-Assman P, Cavalier G, Dracopoli N: Fogh J, et al.: Enterocytic differentiation in cultured human colon cancer cells by replacement of glucose by galactose in the medium. Biol Cell 44: 193–196, 1982

    Google Scholar 

  29. Zweibaum A, Pinto M, Chevalier G, Dussaulx E, Triadou N, Lacroix B, et al.: Enterocytic differentiation of a subpopulation of the human colon cancer cell line HT-29 selected for growth in sugar-free medium and its inhibition by glucose. J Cell Physiol 122: 21–29, 1985

    PubMed  Google Scholar 

  30. Simons SK, Fuller SD: Cell surface polarity in epithelia. Ann Rev Cell Biol 1: 243–288, 1985

    PubMed  Google Scholar 

  31. Rodriguez-Boulon E, Nelson WJ: Morphogenesis of the polarized epithelial cell phenotype. Science 245: 718–725, 1989

    Google Scholar 

  32. Cereijido M, Contreras RG, Gonzalez-Mariscal L: Development and alteration of polarity. Ann Rev Physiol 51: 585–595, 1989

    Google Scholar 

  33. Wang JYJ, Knudsen ES, Welch PJ: The retinoblastoma tumor suppressor protein. Adv Cancer Res 64: 25–85, 1994

    PubMed  Google Scholar 

  34. Almasan A, Yin Y, Kelly RE, EP Lee, Bradley A, Li W, Bertino JR, Wahl GM: Deficiency of retinoblastoma protein leads to inappropriate S-phase entry, activation of E2F-responsive genes, and apoptosis. Proc Natl Acad Sci 92: 5436–5440, 1995

    PubMed  Google Scholar 

  35. Csordas A: On the biological role of histone acetylation. Biochem J 265: 23–38, 1990

    PubMed  Google Scholar 

  36. Herman JG, Latif F, Weng Y, Lerman MI, Zbar B, Liu S, Samid D, et al.: Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma. Proc Natl Acad Sci 91: 9700–9704, 1994

    PubMed  Google Scholar 

  37. Yoshiura K, Kanai Y, Ochiai A, Shimoyama Y, Sugimura T, Hirohashi S: Silencing of the E-cadherin invasion-suppressor gene by CpG methylation in human carcinomas. Proc Natl Acad Sci 92: 7416–7419, 1995

    PubMed  Google Scholar 

  38. Merlo A, Herman JG, Lee DJ, Gabrielson E, Burger PC, Baylin SB, Sidransky D: 5'-CpG island methylation is associated with transcriptional silencing of the tumor suppressor p16/CDKN2/MTS1 in human cancers. Nat Med 1: 686–692, 1995

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schwartz, B., Avivi-Green, C. & Polak-Charcon, S. Sodium butyrate induces retinoblastoma protein dephosphorylation, p16 expression and growth arrest of colon cancer cells. Mol Cell Biochem 188, 21–30 (1998). https://doi.org/10.1023/A:1006831330340

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006831330340

Navigation