1. Volume 52, Issue 6, Pages 1670-1681 (December 2007)
Increased Expression of Tumor-Associated Trypsin Inhibitor, TATI, in Prostate Cancer and in Androgen-Independent 22Rv1 Cells 
Annukka Paju, Kristina Hotakainen, Yue Cao, Timo Laurila, Virgil Gadaleanu, Akseli Hemminki, Ulf-Håkan Stenman, Anders Bjartell 
European Urology 
Volume 52, Issue 6, Pages (December 2007)
DOI: /j.eururo
Copyright © 2007 European Association of Urology Terms and Conditions

2. Fig. 1
Immunohistochemical localization of tumor-associated trypsin inhibitor (TATI) and chromogranin A (CgA) in paraffin sections from benign and malignant prostatic tissues. (A) Tissue microarray (TMA) with a prostatic tumor (Gleason grade 4) with moderate to strong immunoreactivity for TATI in tumor cells (to the left) and weak or not detectable staining in adjacent benign glands (monoclonal antibody 6E8). (B) Prostate cancer (Gleason grade 3) with moderate intensity of TATI immunostaining. Adjacent TMA sections with strong expression of TATI in most tumor cells (C), some of which show neuroendocrine differentiation as indicated by CgA immunostaining (D). Another tumor with neuroendocrine differentiation was stained for TATI (E) with moderately intensity and a major subpopulation of the cells were CgA-positive (F). (G) Negative control experiment. Strong immunostaining for TATI in prostate cancer and adjacent sections devoid of immunostaining after incubation with an overnight incubation of anti-TATI IgGs with TATI antigen in 10× molar excess (H). (A–F,I–J) alkaline phosphatase antialkaline phosphatase (APAAP) technique with Fast Red as chromophore used for TATI immunostainings and SA-HRP with DAB as chromogen. (Original magnification: ×100 (A) and ×400 [B–H].).
European Urology  , DOI: ( /j.eururo )
Copyright © 2007 European Association of Urology Terms and Conditions

3. Fig. 2
Nonradioactive in situ hybridization for the localization of tumor-associated trypsin inhibitor (TATI) and prostate-specific antigen (PSA) messenger RNA (mRNA) in paraffin sections from prostatic tissues and from the pancreas. (A) Hybridization with TATI antisense oligonucleotide shows a staining pattern similar to TATI immunostaining with the most intense staining in basal epithelial cells. Prostate cancer sections hybridized with antisense oligodeoxynucleotide probes to identify TATI mRNA (B), PSA mRNA (C) and negative staining with TATI oligodeoxynucleotide sense probe (D). Control experiment with sections from the pancreas showing strong hybridization signals after hybridization with the TATI antisense probe (E) and no signals detected when the PSA antisense probe was used (F). Alkaline phosphatase-conjugated Fab fragments of anti-digoxigenin IgGs for detection with BCIP-NBT as substrates. (Original magnification ×100 (A) and ×400 [B–F].).
European Urology  , DOI: ( /j.eururo )
Copyright © 2007 European Association of Urology Terms and Conditions

4. Fig. 3
Immunohistochemical staining of tissue microarrays of benign prostatic hyperplasia (BPH) and prostate cancer (Gleason grades 2, 3, 4, and 5) using monoclonal antibodies against tumor-associated trypsin inhibitor (TATI) (A), prostate-specific antigen (PSA) (B), Ki-67 (C), and chromogranin A (CgA) (D). Mean values of staining intensity were retrieved from examination of individual sample spots by using an arbitrary scale 0, 1, 2, and 3 for TATI, PSA and CgA. The scoring system for Ki-67 was based on the fractions of positively staining nuclei. The results represent mean intensities with ±1 standard errors. (A) *One-way analysis of variance (ANOVA, 2-sided) p value for staining intensity in prostate cancers of increasing Gleason grade. **The Kruskal-Wallis nonparametric ANOVA test p value for staining intensity in benign prostate tissue and in prostate cancers of increasing Gleason grade. (B, C, and D). +ANOVA p value for staining intensity in benign prostate tissue and in prostate cancers of increasing Gleason grade.
European Urology  , DOI: ( /j.eururo )
Copyright © 2007 European Association of Urology Terms and Conditions

5. Fig. 4
Association of the TATI and PSA levels in serum of 66 patients with prostate cancer. The clinical characteristics of patients are shown in Table 1.
European Urology  , DOI: ( /j.eururo )
Copyright © 2007 European Association of Urology Terms and Conditions

6. Fig. 5
Effect of synthetic androgen (R1881) on the release of tumor-associated trypsin inhibitor (TATI) (A) and prostate-specific antigen (PSA) (B) protein into culture medium of 22Rv1 cells and on the release of PSA protein (C) into culture medium of LNCaP cells. Cells were grown in the presence of fetal bovine serum (FBS) (22Rv1 cells) or in the absence of FBS (LNCaP cells) and in the absence or presence of 10 and 20nM R1881. After 4 d, TATI and PSA protein level was measured with the use of specific immunoassays, and the protein levels were corrected for the cell number determined with the use of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT)-based cell proliferation assay. The results represent mean values and standard deviations from four (control wells) and six (wells incubated with R1881) parallel wells. Mann-Whitney U test (2-sided).
European Urology  , DOI: ( /j.eururo )
Copyright © 2007 European Association of Urology Terms and Conditions