Cell Free DNA of Tumor Origin Induces a 'Metastatic' Expression Profile in HT-29 Cancer Cell Line

被引:27
作者
Furi, Istvan [2 ]
Kalmar, Alexandra [1 ]
Wichmann, Barnabas [2 ]
Spisak, Sandor [2 ]
Schoeller, Andrea [1 ]
Bartak, Barbara [1 ]
Tulassay, Zsolt [2 ]
Molnar, Bela [2 ]
机构
[1] Semmelweis Univ, Dept Internal Med 2, H-1085 Budapest, Hungary
[2] Hungarian Acad Sci, Mol Med Res Unit, Budapest, Hungary
来源
PLOS ONE | 2015年 / 10卷 / 07期
基金
匈牙利科学研究基金会;
关键词
TOLL-LIKE RECEPTOR-9; NUCLEIC-ACIDS; INVASION; PROMOTES; PROTEIN; PLASMA; TOLL-LIKE-RECEPTOR-9; INFLAMMATION; RECOGNITION; BIOMARKER;
D O I
10.1371/journal.pone.0131699
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background Epithelial cells in malignant conditions release DNA into the extracellular compartment. Cell free DNA of tumor origin may act as a ligand of DNA sensing mechanisms and mediate changes in epithelial-stromal interactions. Aims To evaluate and compare the potential autocrine and paracrine regulatory effect of normal and malignant epithelial cell-related DNA on TLR9 and STING mediated pathways in HT-29 human colorectal adenocarcinoma cells and normal fibroblasts. Materials and Methods DNA isolated from normal and tumorous colonic epithelia of fresh frozen surgically removed tissue samples was used for 24 and 6 hour treatment of HT-29 colon carcinoma and HDF-alpha fibroblast cells. Whole genome mRNA expression analysis and qRT-PCR was performed for the elements/members of TLR9 signaling pathway. Immunocytochemistry was performed for epithelial markers (i.e. CK20 and E-cadherin), DNA methyltransferase 3a (DNMT3a) and NF kappa B (for treated HDFa cells). Results Administration of tumor derived DNA on HT29 cells resulted in significant (p<0.05) mRNA level alteration in 118 genes (logFc >= 1, p <= 0.05), including overexpression of metallothionein genes (i.e. MT1H, MT1X, MT1P2, MT2A), metastasis-associated genes (i.e. TACSTD2, MACC1, MALAT1), tumor biomarker (CEACAM5), metabolic genes (i.e. INSIG1, LIPG), messenger molecule genes (i.e. DAPP, CREB3L2). Increased protein levels of CK20, E-cadherin, and DNMT3a was observed after tumor DNA treatment in HT-29 cells. Healthy DNA treatment affected mRNA expression of 613 genes (logFc >= 1, p <= 0.05), including increased expression of key adaptor molecules of TLR9 pathway (e.g. MYD88, IRAK2, NF kappa B, IL8, IL-1 beta), STING pathway (ADAR, IRF7, CXCL10, CASP1) and the FGF2 gene. Conclusions DNA from tumorous colon epithelium, but not from the normal epithelial cells acts as a prometastatic factor to HT-29 cells through the overexpression of pro-metastatic genes through TLR9/MYD88 independent pathway. In contrast, DNA derived from healthy colonic epithelium induced TLR9 and STING signaling pathway in normal fibroblasts.
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页数:16
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