title: TLR3 expression correlates with apoptosis, proliferation and angiogenesis in hepatocellular carcinoma and predicts prognosis.
abstract: BACKGROUND
  Toll-like receptor 3 (TLR3) plays a key role in innate immunity. In the present study, we analyzed tissues of patients with human hepatocellular carcinoma (HCC) to determine the significance of the relationship between TLR3 expression and cell proliferation, apoptosis, hepatitis B virus infections, angiogenesis and prognosis.
  
  
  METHODS
  We collected paraffin-embedded tissues from 85 patients with HCC who had complete histories and were followed for >5 years. The expression and intracellular localization of TLR3 and downstream proteins (TRIF, NF-κB, and IRF3) were detected using immunohistochemistry. Further, we determined the expression of proteins that mediate cell proliferation (Ki67, cyclin D1), apoptosis (survivin, bcl-2, caspases 3, 8, and 9), and angiogenesis (CD34, MMP-2) as well as the HBV proteins HBsAg and HBcAg. Apoptosis in HCC tissues was detected using TUNEL. We conducted dual-labeling immunohistochemical analyses of TLR3 expression and TUNEL activity.
  
  
  RESULTS
  TLR3 expression was significantly lower in HCC tissues compared with adjacent tissues. TRIF, NF-κB, and IRF3 correlated positively with TLR3 expression. Survivin and Bcl-2 expression correlated negatively with TLR3. The frequencies of caspases 3, 8, and 9 expression correlated positively with TLR3 signaling proteins. Cytoplasmic TLR3 and serum levels of HBsAg correlated positively. The apoptotic index determined using the TUNEL method and correlated positively with TLR3 expression. TLR3 expression in the cytoplasm correlated positively with TUNEL-positive cells and HBsAg. Ki67 and cyclin D1 correlated negatively with TLR3 expression. MMP-2 expression, microvessel density (CD34(+)) and endothelial progenitor cells (EPCs) correlated negatively with TLR3 expression. Kaplan-Meier survival analysis shows that TLR3 expression correlated with longer survival.
  
  
  CONCLUSIONS
  The expression of TLR3 in HCC tissues may exert a synergistic effect on apoptosis and inhibit the proliferation of HCC cells, MMP-2 expression, generation of EPCs, and angiogenesis. Moreover, TLR3 expression may serve as a prognostic marker of HCC.
authors.affiliation: Department of Pathological Anatomy, Nantong University, Qixiu Road 19, Nantong City, Jiangsu, 226001, China. 378842114@qq.com.|Department of Pathology, Nantong Rich Hospital, Jiangsu, China. 378842114@qq.com.
authors.forename: Ming-Ming
authors.initials: MM
authors.lastname: Yuan
authors.affiliation: Department of Nephrology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China. xuyuyin27@aliyun.com.
authors.forename: Yu-Yin
authors.initials: YY
authors.lastname: Xu
authors.affiliation: Department of Pathological Anatomy, Nantong University, Qixiu Road 19, Nantong City, Jiangsu, 226001, China. bl1@ntu.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Pathological Anatomy, Nantong University, Qixiu Road 19, Nantong City, Jiangsu, 226001, China. kawayelily@ntu.edu.cn.
authors.forename: Xing-Yu
authors.initials: XY
authors.lastname: Li
authors.affiliation: Department of Pathological Anatomy, Nantong University, Qixiu Road 19, Nantong City, Jiangsu, 226001, China. qj_5400@163.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Qin
authors.affiliation: Department of Pathological Anatomy, Nantong University, Qixiu Road 19, Nantong City, Jiangsu, 226001, China. podiori@126.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Shen
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D000956:Antigens, Viral / Q000097:blood; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000401:mortality / Q000473:pathology / Q000821:virology; D049109:Cell Proliferation; D015870:Gene Expression; D006509:Hepatitis B / Q000378:metabolism; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D050838:Interferon Regulatory Factor-3 / Q000378:metabolism; D053208:Kaplan-Meier Estimate; D008113:Liver Neoplasms / Q000378:metabolism* / Q000401:mortality / Q000473:pathology / Q000821:virology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009389:Neovascularization, Pathologic; D011379:Prognosis; D021381:Protein Transport; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D000077022:Survivin; D051196:Toll-Like Receptor 3 / Q000378:metabolism*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D018952:Antigens, CD34; D000956:Antigens, Viral; C000595707:BCL2 protein, human; C107489:BIRC5 protein, human; C494232:IRF3 protein, human; D051034:Inhibitor of Apoptosis Proteins; D050838:Interferon Regulatory Factor-3; D016328:NF-kappa B; D019253:Proto-Oncogene Proteins c-bcl-2; D000077022:Survivin; C470409:TICAM1 protein, human; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2