The tumor microenvironment of colorectal cancer: stromal TLR-4 expression as a potential prognostic marker.
BACKGROUND
Colorectal cancer can be efficiently treated when found at early stages, thus the search for novel markers is of paramount importance. Since inflammation is associated with cancer progression and angiogenesis, we investigated expression of cytokines like IL-6 and other mediators that play a key role in the innate immune system, in particular toll like receptor 4 (TLR4), in the microenvironment of lesions from different stages of colon disease progression, from ulcerative colitis to adenoma and adenocarcinoma to find useful markers.


METHODS
The presence of inflammatory cells and expression of key cytokines involved in the inflammation process were quantified by immunohistochemistry in specific tissue compartments (epithelial, stromal, endothelial) by immunohistochemistry. A murine azoxymethane/dextran sulfate model in which Tir8, a negative regulator of the inflammatory response, was ablated was used to confirm the clinical observations. 116 Archival tissue samples from patients with different stages of colorectal disease: 13 cases of ulcerative colitis (UC), 34 tubular or tubulo-villous adenomas (AD), and 53 infiltrating adenocarcinomas. 16 specimens of healthy mucosa surgically removed with the cancerous tissue were used as a control.


RESULTS
The differences between healthy tissues and the diverse lesions was characterized by a marked inflammatory-angiogenic reaction, with significantly (P < 0.05) higher numbers of CD68, CD15, and CD31 expressing cells in all diseased tissues that correlated with increasing grade of malignancy. We noted down-regulation of a potential modulator molecule, Hepatocyte Growth Factor, in all diseased tissues (P < 0.05). TLR-4 and IL6 expression in the tumor microenvironment were associated with adenocarcinoma in human samples and in the murine model. We found that adenocarcinoma patients (pT1-4) with higher TLR-4 expression in stromal compartment had a significantly increased risk in disease progression. In those patients with a diagnosis of pT3 (33 cases) colon cancer, those with very high levels of TLR-4 in the tumor stroma relapsed significantly earlier than those with lower expression levels.


CONCLUSIONS
These data suggest that high TLR-4 expression in the tumor microenvironment represents a possible marker of disease progression in colon cancer.
Oncology Research Laboratory, Science and Technology Park, IRCCS MultiMedica, (via Fantoli 16/15), Milan, (20138), Italy.
Rosaria
R
Cammarota
Valentina
V
Bertolini
Giuseppina
G
Pennesi
Eraldo O
EO
Bucci
Ornella
O
Gottardi
Cecilia
C
Garlanda
Luigi
L
Laghi
Massimo C
MC
Barberis
Fausto
F
Sessa
Douglas M
DM
Noonan
Adriana
A
Albini
D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D015179:Colorectal Neoplasms / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D011379:Prognosis; D051197:Toll-Like Receptor 4 / Q000378:metabolism*
D015703:Antigens, CD; D016207:Cytokines; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4