No abstract

-12). In most of these cases, the sites affected were the lymph nodes. Extranodal cases were uncommon. Approximately 12% of FDC tumors were demonstrated to contain the EpsteinBarr virus (EBV) sequence (7). Therefore, EBV may play a role in the pathogenesis of the FDC tumor. In this report, we describe two FDC tumors of the liver with unique clinical features and a unique EBV subtype. MATERIALS AND METHODSTwo cases of hepatic FDC tumor were retrieved from the surgical pathology files of Chang Gung Memorial Hospital from the period 1996 to 1999. Formalin-fixed and paraffin-embedded tissues were used for histopathologic, immunohistochemical, and in situ hybridization studies. The immunohistochemical studies were performed using the avidin-biotin-peroxidase complex method. A panel of antibodies was used, and the antibodies are listed in Table 1. A fresh skin biopsy specimen (Case 1) was also subjected to direct immunofluorescence studies for IgG, IgM, IgA, C 3 , and C 4 . EBV-encoded nuclear RNA (EBER) in situ hybridization studies were performed with a DAKO fluoresceinconjugated EBV (EBER) PNA probe (complementary to two nuclear EBER RNAs encoded by EBV; DAKO A/S, Glostrup, Denmark) on 5-m-thick, deparaffinized, and proteinase K-pretreated tissues for 1.5 hour at 55°C. After washing, the reaction was detected by the DAKO PNA ISH detection Kit (DAKO A/S). Latent Membrane Protein-1 Gene StudyFresh tumor tissue obtained from Case 2 was frozen at Ϫ80°C until analyzed. High-molecular weight DNA was purified by phenol/chloroform ex-

More than 380,000 new cases of bladder cancer are diagnosed worldwide, accounting for ϳ150,200 deaths each year. To discover potential biomarkers of bladder cancer, we employed a strategy combining laser microdissection, isobaric tags for relative and absolute quantitation labeling, and liquid chromatography-tandem MS (LC-MS/MS) analysis to profile proteomic changes in fresh-frozen bladder tumor specimens. Cellular proteins from four pairs of surgically resected primary bladder cancer tumor and adjacent nontumorous tissue were extracted for use in two batches of isobaric tags for relative and absolute quantitation experiments, which identified a total of 3220 proteins. A DAVID (database for annotation, visualization and integrated discovery) analysis of dysregulated proteins revealed that the three top-ranking biological processes were extracellular matrix organization, extracellular structure organization, and oxidation-reduction. Biological processes including response to organic substances, response to metal ions, and response to inorganic substances were highlighted by up-expressed proteins in bladder cancer. Seven differentially expressed proteins were selected as potential bladder cancer biomarkers for further verification. Immunohistochemical analyses showed significantly elevated levels of three proteins-SLC3A2, STMN1, and TAGLN2-in tumor cells compared with noncancerous bladder epithelial cells, and suggested that TAGLN2 could be a useful tumor tissue marker for diagnosis (AUC ‫؍‬ 0.999) and evaluating lymph node metastasis in bladder cancer patients. ELISA results revealed significantly increased urinary levels of both STMN1 and TAGLN2 in bladder cancer subgroups compared with control groups. In comparisons with agematched hernia urine specimens, urinary TAGLN2 in bladder cancer samples showed the largest fold change (7.13-fold), with an area-under-the-curve value of 0.70 (p < 0.001, n ‫؍‬ 205). Overall, TAGLN2 showed the most significant overexpression in individual bladder cancer tissues and urine specimens, and thus represents a potential biomarker for noninvasive screening for bladder cancer. Our findings highlight the value of bladder tissue proteome in providing valuable information for future validation studies of potential biomarkers in urothelial carcinoma. Molecular & Cellular Proteomics