ABSTRACT The microbial pattern-recognizing Toll-like receptors (TLRs) are major signal transducers known to shape and influence the postnatal maturation of host intestinal epithelium.

Perturbations in this intricate host–microbe cross-talk have been reported to be associated with uncontrolled epithelial cell growth and thus potential cancer development by mechanisms which

are largely unknown. We therefore generated transgenic mice carrying a constitutively active TLR4 (CD4-TLR4) linked to an intestinal epithelial cell-specific promoter. _Ex vivo_ analysis of

transgenic crypt-villus organoid cultures revealed an increased proliferative capacity and a lowered cyclooxygenase 2 (Cox-2) expression in these organoids compared with wild-type control

increased apoptosis _in vivo_. Thus, our data reveal that host microbiota-mediated signal transduction via TLR4 in intestinal epithelial cells is far more complex than what is previously

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MATURATION AND CONTROLS SMALL INTESTINAL IMMUNE CELL COMPOSITION INDEPENDENT OF MICROBIOTA IN A MURINE MODEL Article Open access 22 April 2024 REFERENCES * Rakoff-Nahoum S, Medzhitov R .

Regulation of spontaneous intestinal tumorigenesis through the adaptor protein MyD88. _Science_ 2007; 317: 124–127. Article  CAS  Google Scholar  * Luo J-L, Maeda S, Hsu L-C, Yagita H, Karin

M . Inhibition of NF-κ B in cancer cells converts inflammation- induced tumor growth mediated by TNFα to TRAIL-mediated tumor regression. _Cancer Cell_ 2004; 6: 297–305. Article  CAS 

Google Scholar  * Pidgeon GP, Harmey JH, Kay E, Costa MD, Redmond HP, Bouchier-Hayes DJ . The role of endotoxin/lipopolysaccharide in surgically induced tumour growth in a murine model of

metastatic disease. _Br J Cancer_ 1999; 81: 1311–1317. Article  CAS  Google Scholar  * Harmey JH, Bucana CD, Lu W, Byrne AM, McDonnell S, Lynch C _et al_. Lipopolysaccharide-induced

metastatic growth is associated with increased angiogenesis, vascular permeability and tumor cell invasion. _Int J Cancer_ 2002; 101: 415–422. Article  CAS  Google Scholar  * Chicoine M,

Zahner M, Won E, Kalra R, Kitamura T, Perry A _et al_. The in vivo antitumoral effects of lipopolysaccharide against glioblastoma multiforme are mediated in part by Toll-like receptor 4.

_Neurosurgery_ 2007; 60: 372–380. Article  Google Scholar  * Andreani V, Gatti G, Simonella L, Rivero V, Maccioni M . Activation of Toll-like Receptor 4 on Tumor Cells In vitro Inhibits

Subsequent Tumor Growth In vivo. _Cancer Res_ 2007; 67: 10519–10527. Article  CAS  Google Scholar  * Maehara Y, Okuyama T, Kakeji Y, Baba H, Furusawa M, Sugimachi K . Postoperative

immunochemotherapy including streptococcal lysate OK-432 is effective for patients with gastric cancer and serosal invasion. _Am J Surg_ 1994; 168: 36–40. Article  CAS  Google Scholar  *

Sato M, Harada K, Yoshida H, Yura Y, Azuma M, Iga H _et al_. Therapy for oral squamous cell carcinoma by tegafur and streptococcal agent OK-432 in combination with radiotherapy: association

of the therapeutic effect with differentiation and apoptosis in the cancer cells. _Apoptosis_ 1997; 2: 227–238. Article  CAS  Google Scholar  * Otto F, Schmid P, Mackensen A, Wehr U, Seiz A,

Braun M _et al_. Phase II trial of intravenous endotoxin in patients with colorectal and non-small cell lung cancer. _Eur J Cancer_ 1996; 32A: 1712–1718. Article  CAS  Google Scholar  *

Razack AHA . Bacillus Calmette–Guérin and bladder cancer. _Asian J Surg_ 2007; 30: 302–309. Article  Google Scholar  * Garay RP, Viens P, Bauer J, Normier Gr, Bardou M, Jeannin J-Fo _et al_.

Cancer relapse under chemotherapy: Why TLR2/4 receptor agonists can help. _Eur J Pharmacol_ 2007; 563: 1–17. Article  CAS  Google Scholar  * Smith WL, DeWitt DL, Garavito RM .

CYCLOOXYGENASES: structural, cellular, and molecular biology. _Annu Rev Biochem_ 2000; 69: 145–182. Article  CAS  Google Scholar  * Trifan OC, Hla T . Cyclooxygenase-2 modulates cellular

growth and promotes tumorigenesis. _J Cell Mol Med_ 2003; 7: 207–222. Article  CAS  Google Scholar  * Wang D, DuBois RN . Prostaglandins and cancer. _Gut_ 2006; 55: 115–122. Article  CAS 

Google Scholar  * Steinbach G, Lynch PM, Phillips RKS, Wallace MH, Hawk E, Gordon GB _et al_. The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis. _N

Engl J Med_ 2000; 342: 1946–1952. Article  CAS  Google Scholar  * Swamy MV, Patlolla JMR, Steele VE, Kopelovich L, Reddy BS, Rao CV . Chemoprevention of familial adenomatous polyposis by low

doses of atorvastatin and celecoxib given individually and in combination to APCMin mice. _Cancer Res_ 2006; 66: 7370–7377. Article  CAS  Google Scholar  * Li Y, Kundu P, Seow SW, Teixeira

de Matos C, Linda A, Chin KC _et al_. Gut microbiota accelerate tumor growth via c-Jun and STAT3 phosphorylation in APCMin/+ mice. _Carcinogenesis_ 2012; 33: 1231–1238. Article  CAS  Google

Scholar  * Yokota S-i, Okabayashi T, Rehli M, Fujii N, Amano K-i . Helicobacter pylori lipopolysaccharides upregulate toll-like receptor 4 expression and proliferation of gastric epithelial

cells via the MEK1/2-ERK1/2 mitogen-activated protein kinase pathway. _Infect Immun_ 2010; 78: 468–476. Article  CAS  Google Scholar  * Bohnhorst J, Rasmussen T, Moen SH, Flottum M, Knudsen

L, Borset M _et al_. Toll-like receptors mediate proliferation and survival of multiple myeloma cells. _Leukemia_ 2006; 20: 1138–1144. Article  CAS  Google Scholar  * Moser A, Pitot H, Dove

W . A dominant mutation that predisposes to multiple intestinal neoplasia in the mouse. _Science_ 1990; 247: 322–324. Article  CAS  Google Scholar  * Medzhitov R, Preston-Hurlburt P, Janeway

CA . A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. _Nature_ 1997; 388: 394–397. Article  CAS  Google Scholar  * Sato T, van Es JH, Snippert HJ,

Stange DE, Vries RG, van den Born M _et al_. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. _Nature_ 2011; 469: 415–418. Article  CAS  Google Scholar  *

Gregorieff A, Pinto D, Begthel H, Destrée O, Kielman M, Clevers H . Expression pattern of wnt signaling components in the adult intestine. _Gastroenterology_ 2005; 129: 626–638. Article 

CAS  Google Scholar  * Kargman SL, O’Neill GP, Vickers PJ, Evans JF, Mancini JA, Jothy S . Expression of prostaglandin G/H synthase-1 and -2 protein in human colon cancer. _Cancer Res_ 1995;

55: 2556–2559. CAS  PubMed  Google Scholar  * Kutchera W, Jones DA, Matsunami N, Groden J, McIntyre TM, Zimmerman GA _et al_. Prostaglandin H synthase 2 is expressed abnormally in human

colon cancer: evidence for a transcriptional effect. _Proc Natl Acad Sci_ 1996; 93: 4816–4820. Article  CAS  Google Scholar  * Sheng H, Shao J, Kirkland SC, Isakson P, Coffey RJ, Morrow J

_et al_. Inhibition of human colon cancer cell growth by selective inhibition of cyclooxygenase-2. _J Clin Invest_ 1997; 99: 2254–2259. Article  CAS  Google Scholar  * Núňez NG, Andreani V,

Crespo MI, Nocera DA, Breser ML, Morón G _et al_. IFNβ produced by TLR4-activated tumor cells is involved in improving the antitumoral immune response. _Cancer Res_ 2012; 72: 592–603.

Article  Google Scholar  * Cursio R, Colosetti P, Auberger P, Gugenheim J . Liver apoptosis following normothermic ischemia-reperfusion: in vivo evaluation of caspase activity by FLIVO assay

in rats. _Transplant Proc_ 2008; 40: 2038–2041. Article  CAS  Google Scholar  * Fukata M, Shang L, Santaolalla R, Sotolongo J, Pastorini C, España C _et al_. Constitutive activation of

epithelial TLR4 augments inflammatory responses to mucosal injury and drives colitis-associated tumorigenesis. _Inflamm Bowel Dis_ 2011; 17: 1464–1473. Article  Google Scholar  * Cohn SM,

Simon TC, Roth KA, Birkenmeier EH, Gordon JI . Use of transgenic mice to map cis-acting elements in the intestinal fatty acid binding protein gene (FABPI) that control its cell

lineage-specific and regional patterns of expression along the duodenal-colonic and crypt-villus axes of the gut epithelium. _J Cell Biol_ 1992; 119: 27–44. Article  CAS  Google Scholar  *

Sweetser D, Hauft S, Hoppe P, Birkenmeier E, Gordon J . Transgenic mice containing intestinal fatty acid-binding protein-human growth hormone fusion genes exhibit correct regional and

cell-specific expression of the reporter gene in their small intestine. _Proc Natl Acad Sci USA_ 1988; 85: 9611–9615. Article  CAS  Google Scholar  * Matsushita K, Takeuchi O, Standley DM,

Kumagai Y, Kawagoe T, Miyake T _et al_. Zc3h12a is an RNase essential for controlling immune responses by regulating mRNA decay. _Nature_ 2009; 458: 1185–1190. Article  CAS  Google Scholar 

* Fukata M, Chen A, Klepper A, Krishnareddy S, Vamadevan AS, Thomas LS _et al_. Cox-2 is regulated by toll-like receptor-4 (TLR4) signaling: role in proliferation and apoptosis in the

intestine. _Gastroenterology_ 2006; 131: 862–877. Article  CAS  Google Scholar  * Brown SL, Riehl TE, Walker MR, Geske MJ, Doherty JM, Stenson WF _et al_. Myd88-dependent positioning of

Ptgs2-expressing stromal cells maintains colonic epithelial proliferation during injury. _J Clin Invest_ 2007; 117: 258–269. Article  CAS  Google Scholar  * Singh B, Berry JA, Shoher A,

Ramakrishnan V, Lucci A . COX-2 overexpression increases motility and invasion of breast cancer cells. _Int J Oncol_ 2005; 26: 1393–1399. CAS  PubMed  Google Scholar  * Müller-Decker K,

Neufang G, Berger I, Neumann M, Marks F, Fürstenberger G . Transgenic cyclooxygenase-2 overexpression sensitizes mouse skin for carcinogenesis. _Proc Natl Acad Sci_ 2002; 99: 12483–12488.

Article  Google Scholar  * Nakamoto N, Higuchi H, Kanamori H, Kurita S, Tada S, Takaishi H _et al_. Cyclooxygenase-2 inhibitor and interferon-β synergistically induce apoptosis in human

hepatoma cells in vitro and in vivo. _Int J Oncol_ 2006; 29: 625–635. CAS  PubMed  Google Scholar  * Klampfer L, Huang J, Kaler P, Sasazuki T, Shirasawa S, Augenlicht L . STAT1-independent

inhibition of cyclooxygenase-2 expression by IFNγ; a common pathway of IFNγ-mediated gene repression but not gene activation. _Oncogene_ 2007; 26: 2071–2081. Article  CAS  Google Scholar  *

Ohtsuka Y, Lee J, Stamm DS, Sanderson IR . MIP-2 secreted by epithelial cells increases neutrophil and lymphocyte recruitment in the mouse intestine. _Gut_ 2001; 49: 526–533. Article  CAS 

Google Scholar  * Arulampalam V, Grant PA, Samuelsson A, Lendahl U, Pettersson S . Lipopolysaccharide-dependent transactivation of the temporally regulated immunoglobulin heavy chain 3’

enhancer. _Eur J Immunol_ 1994; 24: 1671–1677. Article  CAS  Google Scholar  * Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE _et al_. Single Lgr5 stem cells build

crypt–villus structures in vitro without a mesenchymal niche. _Nature_ 2009; 459: 262–265. Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS Our research work was funded by

a grant from Singapore Immunology Network (SIgN)—SIgN 10-038. We would like to thank Dr Robert Wallin for his great expertise and assistance in using the deconvolution microscope for

anti-CD4 fluorescence imaging. We thank Velmurugesan Arulampalam for valuable discussions regarding our study. We also thank Arlaine Anne Bautista Amoyo, Jieshun Wong and Annika Samuelsson

for their technical support with animal care and handling. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institute,

Stockholm, Sweden Y Li, L Meijer & S Pettersson * Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), Singapore Y Li * National Cancer Centre,

Singapore W L Teo, M J Low, S Pettersson & G Greicius * Digestive Diseases Clinical Academic Unit, Barts and The London, Queen Mary’s School of Medicine and Dentistry, London, UK I

Sanderson * School of Biological Sciences, Nanyang Technological University, Singapore S Pettersson Authors * Y Li View author publications You can also search for this author inPubMed 

Google Scholar * W L Teo View author publications You can also search for this author inPubMed Google Scholar * M J Low View author publications You can also search for this author inPubMed 

Google Scholar * L Meijer View author publications You can also search for this author inPubMed Google Scholar * I Sanderson View author publications You can also search for this author

inPubMed Google Scholar * S Pettersson View author publications You can also search for this author inPubMed Google Scholar * G Greicius View author publications You can also search for this

author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to S Pettersson. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no conflict of interest. ADDITIONAL

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permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Li, Y., Teo, W., Low, M. _et al._ Constitutive TLR4 signalling in intestinal epithelium reduces tumor load by increasing apoptosis in

APCMin/+ mice. _Oncogene_ 33, 369–377 (2014). https://doi.org/10.1038/onc.2012.581 Download citation * Received: 26 June 2012 * Revised: 08 October 2012 * Accepted: 26 October 2012 *

Published: 14 January 2013 * Issue Date: 16 January 2014 * DOI: https://doi.org/10.1038/onc.2012.581 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this

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KEYWORDS * TLR4 * APCMin/+ * Cox-2 * apoptosis * Paneth cells * tumorigenesis