Short-Chain Fatty Acid Butyrate Induces Cilia Formation and Potentiates the Effects of HDAC6 Inhibitors in Cholangiocarcinoma Cells

Cholangiocarcinoma (CCA) is a deadly form of liver cancer with limited therapeutic approaches. The pathogenesis of CCA involves the loss of primary cilia in cholangiocytes, an important organelle that regulates several key cellular functions including the regulation of cell polarity, growth, and differentiation, by a mechanism involving increased expression of deacetylases like HDAC6 and SIRT1. Therefore, cilia restoration may represent an alternative and novel therapeutic approach against CCA. Butyrate is produced by bacterial fermentation of fibers in the intestine and has been shown to inhibit SIRT1, showing antitumor effects on various cancers. Herein, we investigated the role of butyrate on CCA cell proliferation, migration, and EMT and evaluated the synergistic effects with specific HDAC6 inhibition. When CCA cells, including HuCCT1 and KMCH, were treated with butyrate, the cilia formation and acetylated-tubulin levels were increased, while no significant effects were observed in normal human cholangiocytes. Butyrate treatment also depicted reduced cell proliferation in HuCCT1 and KMCH cells, but on the other hand, it affected cell growth of the normal cholangiocytes only at high concentrations. In HuCCT1 cells, spheroid formation and cell migration were also halted by butyrate treatment. Furthermore, we found that butyrate augmented the previously described effects of HDAC6 inhibitors on CCA cell proliferation and migration by reducing the expression of CD44, cyclin D1, PCNA, Zeb1, and Vimentin. In summary, butyrate targets cancer cell growth and migration and enhances the anti-cancer effects of HDAC6 inhibitors in CCA cells, suggesting that butyrate may have therapeutic effects in CCA and other ciliopathies.

[1]  N. Giama,et al.  Histone Deacetylase Sirtuin 1 Promotes Loss of Primary Cilia in Cholangiocarcinoma , 2021, Hepatology.

[2]  R. Cowles,et al.  Butyrate and the Intestinal Epithelium: Modulation of Proliferation and Inflammation in Homeostasis and Disease , 2021, Cells.

[3]  S. Pierdomenico,et al.  Epithelial-Mesenchymal Transition (EMT): The Type-2 EMT in Wound Healing, Tissue Regeneration and Organ Fibrosis , 2021, Cells.

[4]  G. Gores,et al.  Cholangiocarcinoma 2020: the next horizon in mechanisms and management , 2020, Nature Reviews Gastroenterology & Hepatology.

[5]  Wenqi Wang,et al.  Sodium Butyrate Selectively Kills Cancer Cells and Inhibits Migration in Colorectal Cancer by Targeting Thioredoxin-1 , 2020, OncoTargets and therapy.

[6]  A. Ballabio,et al.  HDAC6-dependent ciliophagy is involved in ciliary loss and cholangiocarcinoma growth in human cells and murine models. , 2020, American journal of physiology. Gastrointestinal and liver physiology.

[7]  S. Gradilone,et al.  The primary cilium: its role as a tumor suppressor organelle. , 2020, Biochemical pharmacology.

[8]  S. Gradilone,et al.  Role of Histone Deacetylases in Carcinogenesis: Potential Role in Cholangiocarcinoma , 2020, Cells.

[9]  B. Yousefi,et al.  Importance of the Microbiota Inhibitory Mechanism on the Warburg Effect in Colorectal Cancer Cells , 2019, Journal of Gastrointestinal Cancer.

[10]  A. Saraya,et al.  Butyrate inhibits HBV replication and HBV‐induced hepatoma cell proliferation via modulating SIRT‐1/Ac‐p53 regulatory axis , 2018, Molecular carcinogenesis.

[11]  S. Gradilone,et al.  The Chemosensory Function of Primary Cilia Regulates Cholangiocyte Migration, Invasion, and Tumor Growth , 2019, Hepatology.

[12]  Ming-ming Cao,et al.  Butyrate inhibits the proliferation and induces the apoptosis of colorectal cancer HCT116 cells via the deactivation of mTOR/S6K1 signaling mediated partly by SIRT1 downregulation. , 2019, Molecular medicine reports.

[13]  N. LaRusso,et al.  MicroRNA (miR)‐433 and miR‐22 dysregulations induce histone‐deacetylase‐6 overexpression and ciliary loss in cholangiocarcinoma , 2018, Hepatology.

[14]  M. Iwano,et al.  A short-chain fatty acid, propionate, enhances the cytotoxic effect of cisplatin by modulating GPR41 signaling pathways in HepG2 cells , 2018, Oncotarget.

[15]  H. Hao,et al.  Butyrate Suppresses the Proliferation of Colorectal Cancer Cells via Targeting Pyruvate Kinase M2 and Metabolic Reprogramming * , 2018, Molecular & Cellular Proteomics.

[16]  Q. Dong,et al.  Sodium Butyrate Inhibits Colorectal Cancer Cell Migration by Downregulating Bmi‐1 Through Enhanced miR‐200c Expression , 2018, Molecular nutrition & food research.

[17]  M. Casal,et al.  The Role of Diet Related Short-Chain Fatty Acids in Colorectal Cancer Metabolism and Survival: Prevention and Therapeutic Implications. , 2018, Current medicinal chemistry.

[18]  H. Hao,et al.  Butyrate suppresses motility of colorectal cancer cells via deactivating Akt/ERK signaling in histone deacetylase dependent manner. , 2017, Journal of pharmacological sciences.

[19]  A. Saraya,et al.  Oxidative stress plays a key role in butyrate-mediated autophagy via Akt/mTOR pathway in hepatoma cells. , 2017, Chemico-biological interactions.

[20]  N. LaRusso,et al.  Primary Cilia in Tumor Biology: The Primary Cilium as a Therapeutic Target in Cholangiocarcinoma. , 2017, Current drug targets.

[21]  A. Saraya,et al.  Butyrate induces ROS-mediated apoptosis by modulating miR-22/SIRT-1 pathway in hepatic cancer cells , 2017, Redox biology.

[22]  G. Gores,et al.  SOX17 regulates cholangiocyte differentiation and acts as a tumor suppressor in cholangiocarcinoma. , 2017, Journal of hepatology.

[23]  Ansar Karimian,et al.  Multiple functions of p21 in cell cycle, apoptosis and transcriptional regulation after DNA damage. , 2016, DNA repair.

[24]  M. Stiborová,et al.  The synergistic effects of DNA-damaging drugs cisplatin and etoposide with a histone deacetylase inhibitor valproate in high-risk neuroblastoma cells. , 2015, International journal of oncology.

[25]  V. Godfrey,et al.  A gnotobiotic mouse model demonstrates that dietary fiber protects against colorectal tumorigenesis in a microbiota- and butyrate-dependent manner. , 2014, Cancer discovery.

[26]  Harry J. Flint,et al.  The gut microbiota, bacterial metabolites and colorectal cancer , 2014, Nature Reviews Microbiology.

[27]  C. Chiaro,et al.  Butyrate induced changes in Wnt-signaling specific gene expression in colorectal cancer cells , 2014, BMC Research Notes.

[28]  G. Gores,et al.  Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma. , 2014, Journal of hepatology.

[29]  Zhuo Gao,et al.  Sodium Butyrate Induces Growth Inhibition and Apoptosis in Human Prostate Cancer DU145 Cells by Up-Regulation of the Expression of Annexin A1 , 2013, PloS one.

[30]  W. D. de Jonge,et al.  Dietary Inhibitors of Histone Deacetylases in Intestinal Immunity and Homeostasis , 2013, Front. Immunol..

[31]  Nicholas F LaRusso,et al.  HDAC6 inhibition restores ciliary expression and decreases tumor growth. , 2013, Cancer research.

[32]  A. Donner Epigenetics: The multiple HATs of butyrate , 2012 .

[33]  Wei Sun,et al.  The Warburg effect dictates the mechanism of butyrate-mediated histone acetylation and cell proliferation. , 2012, Molecular cell.

[34]  B. Corfe Hypothesis: butyrate is not an HDAC inhibitor, but a product inhibitor of deacetylation. , 2012, Molecular bioSystems.

[35]  J. Oh,et al.  The histone deacetylase inhibitor trichostatin A synergistically resensitizes a cisplatin resistant human bladder cancer cell line. , 2011, The Journal of urology.

[36]  Ying Sun,et al.  Butyrate induces cell apoptosis through activation of JNK MAP kinase pathway in human colon cancer RKO cells. , 2010, Chemico-biological interactions.

[37]  W. Marshall Cilia self-organize in response to planar cell polarity and flow , 2010, Nature Cell Biology.

[38]  Narendra Singh,et al.  Synergistic cytotoxicity of artemisinin and sodium butyrate on human cancer cells. , 2005, Anticancer research.

[39]  Huawei Zeng,et al.  Prolonged butyrate treatment inhibits the migration and invasion potential of HT1080 tumor cells. , 2005, The Journal of nutrition.

[40]  Jorge A. Almenara,et al.  Evidence of a functional role for p21WAF1/CIP1 down-regulation in synergistic antileukemic interactions between the histone deacetylase inhibitor sodium butyrate and flavopiridol. , 2004, Molecular pharmacology.

[41]  R. Toillon,et al.  P21WAF1/CIP1 is dispensable for G1 arrest, but indispensable for apoptosis induced by sodium butyrate in MCF-7 breast cancer cells , 2004, Oncogene.

[42]  B. Evers,et al.  Butyrate inhibits pancreatic cancer invasion , 2003, Journal of Gastrointestinal Surgery.