Butyrate Stimulates Cyclin D and p21 and Inhibits Cyclin-Dependent Kinase 2 Expression in HT-29 Colonic Epithelial Cells

doi:10.1006/bbrc.1997.6255

Biochemical and Biophysical Research Communications

Volume 232, Issue 1, 6 March 1997, Pages 169-172

https://doi.org/10.1006/bbrc.1997.6255 Get rights and content

Abstract

Sodium butyrate, a product of colonic fermentation of dietary fiber, has been shown to inhibit cell proliferation by blocking the cells in the G1 phase of the cell cycle. However, its mechanism of action is still unknown. We found that butyrate strongly stimulated cyclin D and p21/WAF1/CIP1 expression in HT-29 human colonic adenocarcinoma cells, in a dose dependent manner. These stimulations were associated with a decrease in cyclin-dependent kinase (cdk) 2 level, whereas cdk4 and cdk6 remained unchanged. Our results suggest that the inhibition of cell cycle progression by sodium butyrate may be explained by a modulation of cell cycle regulatory proteins such as cyclin D and p21.

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An important function of the gut microbiome is the fermentation of non-digestible dietary fibers into short chain fatty acids (SCFAs). The three primary SCFAs: acetate, propionate, and butyrate, are key mediators of metabolism and immune cell function in the gut mucosa. We previously demonstrated that butyrate at high concentrations decreased human gut lamina propria (LP) CD4 T cell activation in response to enteric bacteria exposure in vitro. However, to date, the mechanism by which butyrate alters human gut LP CD4 T cell activation remains unknown. In this current study, we sought to better understand how exposure to SCFAs across a concentration range impacted human gut LP CD4 T cell function and activation. LP CD4 T cells were directly activated with T cell receptor (TCR) beads in vitro in the presence of a physiologic concentration range of each of the primary SCFAs. Exposure to butyrate potently inhibited CD4 T cell activation, proliferation, and cytokine (IFNγ, IL-17) production in a concentration dependent manner. Butyrate decreased the proliferation and cytokine production of T helper (Th) 1, Th17 and Th22 cells, with differences noted in the sensitivity of LP versus peripheral blood Th cells to butyrate’s effects. Higher concentrations of propionate and acetate relative to butyrate were required to inhibit CD4 T cell activation and proliferation. Butyrate directly increased the acetylation of both unstimulated and TCR-stimulated CD4 T cells, and apicidin, a Class I histone deacetylase inhibitor, phenocopied butyrate’s effects on CD4 T cell proliferation and activation. GPR43 agonism phenocopied butyrate’s effect on CD4 T cell proliferation whereas a GPR109a agonist did not. Our findings indicate that butyrate decreases in vitro human gut LP CD4 T cell activation, proliferation, and inflammatory cytokine production more potently than other SCFAs, likely through butyrate’s ability to increase histone acetylation, and potentially via signaling through GPR43. These findings have relevance in furthering our understanding of how perturbations of the gut microbiome alter local immune responses in the gut mucosa.
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NaB has shown promise as a suitable chemotherapeutic agent due to its myriad anti-tumorigenic effects, including cell proliferation inhibition, induction of cell cycle arrest and differentiation, promotion of apoptosis, and even more significantly as a potent HDAC inhibitor inducing histone hyperacetylation and altering gene expression [6,8,30,31]. Sodium butyrate has been shown to induce p21 expression that is a prominent cell cycle regulatory protein [32,33] In the current study we showed that the potent anti-cancer activities of EGCG and NaB in RKO, HCT-116 and HT-29 CRC cells.
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^☆: J. H. CummingsJ. L. RombeauT. Sakata, Eds.

¹: Address request for reprints to Hervé M. Blottière, INRA, BP 71627, rue de la Géraudière, 44316 Nantes Cedex 03, France. Fax: (33) 02-40-67-50-12; e-mail: [email protected].

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Regular ArticleButyrate Stimulates Cyclin D and p21 and Inhibits Cyclin-Dependent Kinase 2 Expression in HT-29 Colonic Epithelial Cells☆

Abstract

Exp. Cell Res.

Cell

FEBs lett.

Cell

Cell

Physiological and Clinical Aspects of Short Chain Fatty Acids

Int. J. Cancer

Cytometry

Nature

Gene Dev.

Science

Regular Article
Butyrate Stimulates Cyclin D and p21 and Inhibits Cyclin-Dependent Kinase 2 Expression in HT-29 Colonic Epithelial Cells☆