Aberrant Expression of NF-κB in Liver Fluke Associated Cholangiocarcinoma: Implications for Targeted Therapy

Background: Up-regulation and association of nuclear factor kappa B (NF-kappa B) with carcinogenesis and tumor progression has been reported in several malignancies. In the current study, expression of NF-kappa B in cholangiocarcinoma (CCA) patient tissues and its clinical significance were determined. The possibility of using NF-kappa B as the therapeutic target of CCA was demonstrated. Methodology: Expression of NF-kappa B in CCA patient tissues was determined using immunohistochemistry. Dehydroxymethylepoxyquinomicin (DHMEQ), a specific NF-kappa B inhibitor, was used to inhibit NF-kappa B action. Cell growth was determined using an MTT assay, and cell apoptosis was shown by DNA fragmentation, flow cytometry and immunocytofluorescent staining. Effects of DHMEQ on growth and apoptosis were demonstrated in CCA cell lines and CCA-inoculated mice. DHMEQ-induced apoptosis in patient tissues using a histoculture drug response assay was quantified by TUNEL assay. Principal Findings: Normal bile duct epithelia rarely expressed NF-kappa B (subunits p50, p52 and p65), whereas all CCA patient tissues (n = 48) over-expressed all NF-kB subunits. Inhibiting NF-kappa B action by DHMEQ significantly inhibited growth of human CCA cell lines in a dose-and time-dependent manner. DHMEQ increased cell apoptosis by decreasing the antiapoptotic protein expressions-Bcl-2, XIAP-and activating caspase pathway. DHMEQ effectively reduced tumor size in CCA-inoculated mice and induced cell apoptosis in primary histocultures of CCA patient tissues. Conclusions: NF-kappa B was over-expressed in CCA tissues. Inhibition of NF-kappa B action significantly reduced cell growth and enhanced cell apoptosis. This study highlights NF-kappa B as a molecular target for CCA therapy.