Expression of Toll-like receptor 9 increases with progression of cervical neoplasia in Tunisian women--a comparative analysis of condyloma, cervical intraepithelial neoplasia and invasive carcinoma.

Emna Fehri,Samir Boubaker,Monia Ardhaoui,Emna Ennaifer,Kaouther Ouerhani,Ikram Guizani,Rahima Bel Haj Rhouma,Thalja Laassili

doi:10.7314/apjcp.2014.15.15.6145

Emna Fehri, Samir Boubaker + Show 6 more

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https://doi.org/10.7314/apjcp.2014.15.15.6145

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Abstract

Toll-like receptors (TLRs) are expressed in immune and tumor cells and recognize pathogen-associated molecular patterns. Cervical cancer (CC) is directly linked to a persistent infection with high risk human papillomaviruses (HR-HPVs) and could be associated with alteration of TLRs expression. TLR9 plays a key role in the recognition of DNA viruses and better understanding of this signaling pathway in CC could lead to the development of novel immunotherapeutic approaches. The present study was undertaken to determine the level of TLR9 expression in cervical neoplasias from Tunisian women with 53 formalin-fixed and paraffin-embedded specimens, including 22 samples of invasive cervical carcinoma (ICC), 18 of cervical intraepithelial neoplasia (CIN), 7 of condyloma and 6 normal cervical tissues as control cases. Quantification of TLR9 expression was based on scoring four degrees of extent and intensity of immunostaining in squamous epithelial cells. TLR9 expression gradually increased from CIN1 (80% weak intensity) to CIN2 (83.3% moderate), CIN3 (57.1% strong) and ICC (100% very strong). It was absent in normal cervical tissue and weak in 71.4% of condyloma. The mean scores of TLR9 expression were compared using the Kruskall-Wallis test and there was a statistical significance between normal tissue and condyloma as well as between condyloma, CINs and ICC. These results suggest that TLR9 may play a role in progression of cervical neoplasia in Tunisian patients and could represent a useful biomarker for malignant transformation of cervical squamous cells.

Highlights

Cervical cancer (CC) is the second most common malignancy in Tunisia and a public health inquiry in women worldwide
TLR9 is an important receptor of recognition against DNA viruses (Zolini et al, 2014) and a first study was consistent with the lack of relation between Human Papillomaviruses (HPVs) and TLR9 expression and function (Andersen et al, 2006), it has been later demonstrated to be expressed in HPVpositive cervical neoplasia on human foreskin, vaginal and cervical keratinocytes cell lines maintaining episomal copies of HPV16 and 18, in several CC derived cell lines and in formalin-fixed paraffin-embedded or frozen cervical tissue sections (Karim et al, 2011; Werner et al, 2012)
Immunohistochemical scoring of TLR9 staining using a monoclonal antibody has shown that TLR9 expression is detected in all condyloma, cervical intraepithelial neoplasia (CIN) and invasive squamous cells cervical carcinoma (ISCC) with a significant gradual increasing level of expression from CIN1 to ISCC

Summary

Introduction

Cervical cancer (CC) is the second most common malignancy in Tunisia and a public health inquiry in women worldwide. Most of the previous reports have focused on their incubated with an anti-TLR9 monoclonal antibody expression and function in cells of the immune system like (Imgenex Cat.No IMG-305A, clone 26C593) at a 1/250. Current dilution for 45 min at room temperature, washed in PBS works are focusing on their association with cancers and incubated in post- primary Block buffer (Novocastra (Domingos-Pereira et al, 2013; Bodelon et al, 2014) Leica Microsystems) for 30 minutes. Sections were and the use of vaccine with TLR agonist or antagonist in rinsed in 1×Tris and incubated with Novocastra polymer cervical cancer is undergoing extensive investigations for 30 min at room temperature. ISCC using normal cervical tissues and condyloma as control cases

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