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LL‐37/CpG oligonucleotide complex formation and its effects on prostate cancer progression
Author(s) -
Gupta Angela Satya,
Craig Maria Lee,
Deeble Paul D
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.782.2
Subject(s) - electrophoretic mobility shift assay , tlr9 , lncap , prostate cancer , cpg site , microbiology and biotechnology , toll like receptor 9 , cancer research , cancer cell , chemistry , biology , cancer , dna methylation , transcription factor , biochemistry , gene , genetics , gene expression
LL‐37 is an antimicrobial DNA‐binding peptide that has been implicated in anti‐self DNA and anti‐tumor immune responses. The interaction between LL‐37 and DNA is not well‐characterized. We have used electrophoretic mobility shift assays (EMSA) and western blotting to investigate the interaction between LL‐37 and unmethylated CpG sequences known to activate Toll‐like receptor 9 (TLR9) pathways. TLR9 activation by CpG promotes weak anti‐tumor T H 1 immune responses that are enhanced by LL‐37. In addition, we have tested the effects of LL‐37/CpG complexes in a prostate cancer progression model. By EMSA, we observe that LL‐37 is aggregated (has low gel mobility) in the absence of DNA, but that LL‐37/DNA association occurs with increased LL‐37 mobility, suggesting that CpG binding disrupts LL‐37 aggregation. Increases in LL‐37 concentration lead to reduced mobility, indicative of further complexation. We previously found that CpG/LL‐37 increases prostate cancer cell growth (PC‐3 cell line) and invasion through a matrigel matrix (LNCaP cell line). Currently, we are testing the efficacy of CpG/LL‐37 treatments on migration and anchorage‐independent growth in a prostate cancer progression model (PC‐3 cell line) using a cell scratch wound assay and soft agar assay. This work was supported by NIH EARDA grant 5G11HD037064‐08 and a Virginia Foundation of Independent Colleges Summer Undergraduate Research Fellowship.