Non-cancerous Regions Research Articles

Satellite alpha transcripts as a biomarker to predict the development of breast cancer with bilateral or multiple primary malignancies.

e23202 Background: The impaired stability of chromosomal structure promotes the abnormal segregation of chromosomes, increasing the risk of carcinogenesis. Chromosomal stability during segregation is managed by appropriate methylation in the centromeric region of chromosomes. Satellite alpha transcripts (SatA) is a non-coding RNA transcribed from the centromere, which could be involved in this machinery. We reported that overexpression of SatA led to chromosomal instability, which was observed in not only cancer regions (CR) but also noncancerous regions (NCR). It was enhanced in NCR of patients with multiple primary malignancies (MPM), suggesting the involvement of SatA in genomic damage in regions where multiple cancers may arise. In this study, we determined whether SatA is involved in breast cancer development with bilateral or MPM. Methods: We constructed lentiviral vectors expressing SatA and control viruses, which were used to infect human mammary epithelial cells. SatA levels were determined by real-time quantitative PCR (qRT-PCR) and copy number alterations were evaluated by array comparative genomic hybridization (CGH). Then, we investigated the levels of SatA in clinical specimens in 135 breast cancer (BC) patients including 128 tumors and 83 matched normal tissues. Results: qRT-PCR showed higher expression levels of SatA in cells infected with SatA virus than in the control. Array CGH analysis indicated frequent copy number alterations at several specific chromosomes in infected cells, despite no alterations in the control. Among the 135 patients, 24 had bilateral breast cancer (BBC) and 18 had MPM. SatA was much higher in CR in BBC and in MPM than in SBC. SatA was also higher in NCR in MPM than without MPM. Considering human epidermal growth factor receptor type2 (HER2), hormone receptor (HR), family history and other factors, the levels of SatA in CR were a significant independent factor predicting the development of BBC patients. Conclusions: Chromosomal instability induced by SatA is involved in the development of BBC and BC with MPM.

Characterization of a novel androgen receptor (AR) coregulator RIPK1 and related chemicals that suppress AR-mediated prostate cancer growth via peptide and chemical screening.

Using bicalutamide-androgen receptor (AR) DNA binding domain-ligand binding domain as bait, we observed enrichment of FxxFY motif-containing peptides. Protein database searches revealed the presence of receptor-interacting protein kinase 1 (RIPK1) harboring one FxxFY motif. RIPK1 interacted directly with AR and suppressed AR transactivation in a dose-dependent manner. Domain mapping experiments showed that the FxxFY motif in RIPK1 is critical for interactions with AR and the death domain of RIPK1 plays a crucial role in its inhibitory effect on transactivation. In terms of tissue expression, RIPK1 levels were markedly higher in benign prostate hyperplasia and non-cancerous tissue regions relative to the tumor area. With the aid of computer modeling for screening of chemicals targeting activation function 2 (AF-2) of AR, we identified oxadiazole derivatives as good candidates and subsequently generated a small library of these compounds. A number of candidates could effectively suppress AR transactivation and AR-related functions in vitro and in vivo with tolerable toxicity via inhibiting AR-peptide, AR-coregulator and AR N-C interactions. Combination of these chemicals with antiandrogen had an additive suppressive effect on AR transcriptional activity. Our collective findings may pave the way in creating new strategies for the development and design of anti-AR drugs.

Secondary Ion Mass Spectrometry Analysis of Renal Cell Carcinoma with Electrospray Droplet Ion Beams.

Tissue samples from renal cell carcinoma patients were analyzed by electrospray droplet ion beam-induced secondary ion mass spectrometry (EDI/SIMS). Positively- and negatively-charged secondary ions were measured for the cancerous and noncancerous regions of the tissue samples. Although specific cancerous species could not be found in both the positive and negative secondary ion spectra, the spectra of the cancerous and noncancerous tissues presented different trends. For instance, in the m/z range of 500–800 of the positive secondary ion spectra for the cancerous tissues, the intensities for several m/z values were lower than those of the m/z+2 peaks (indicating one double bond loss for the species), whereas, for the noncancerous tissues, the inverse trend was obtained. The tandem mass spectrometry (MS/MS) was also performed on the tissue samples using probe electrospray ionization (PESI), and some molecular ions produced by PESI were found to be fragmented into the ions observed in EDI/SIMS analysis. When the positive secondary ion spectra produced by EDI/SIMS were analyzed by principal component analysis, the results for cancerous and noncancerous tissues were separated. The EDI/SIMS method can be applied to distinguish between a cancerous and a noncancerous area with high probability.

Metabolomic profiling of prostate cancer by matrix assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry imaging using Matrix Coating Assisted by an Electric Field (MCAEF)

In this work, we combined the use of two MALDI matrices (quercetin and 9-aminoacridine), a recently developed new matrix coating technique – matrix coating assisted by an electric field (MCAEF), and matrix-assisted laser desorption/ionization – Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) to detect and image endogenous compounds in the cancerous and non-cancerous regions of three human prostate cancer (stage II) tissue specimens. After three rounds of imaging data acquisitions (i.e., quercetin for positive and negative ion detection and 9-aminoacridine for negative ion detection), and metabolite identification, a total of 1091 metabolites including 1032 lipids and 59 other metabolites were routinely detected and successfully localized. Of these compounds, 250 and 217 were only detected in either the cancerous or the non-cancerous regions respectively, although we cannot rule out the presence of these metabolites at concentrations below the detection limit. In addition, 152 of the other 624 metabolites showed differential distributions (p<0.05, t-test) between the two regions of the tissues. Further studies on a larger number of clinical specimens will need to be carried out to confirm this large number of apparently cancer-related metabolites. The successful determination of the spatial locations and abundances of these endogenous biomolecules indicated significant metabolism abnormalities – e.g., increased energy charge and under-expression of neutral acyl glycerides, in the prostate cancer samples. To our knowledge, this work has resulted in MALDI-MS imaging of the largest group of metabolites in prostate cancer thus far and demonstrated the importance of using complementary matrices for comprehensive metabolomic imaging by MALDI-MS. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann.

Micrometastasis Detection Guidance by Whole-Slide Image Texture Analysis in Colorectal Lymph Nodes

Introduction/ Background Cancer is a disease that affects millions worldwide and accurate determination of whether lymph nodes (LNs) near the primary tumor contain metastatic foci is of critical importance for proper patient management. Histopathological evaluation is the only accepted method to make that determination. However, the current standard of care only examines a single central histological section per LN and yields an unacceptable false-negative rate. Aims To help pathologists in their examination we propose a method that extracts textural features from histopathological LN whole slide images (WSI) and then applies support vector machines (SVMs) to automatically identify regions suspicious for metastatic foci. Methods The database consisted of WSI from 44 LNs. Sections were stained with hematoxylin-eosin and examined at 20x (0.45μm resolution). Twenty-eight of the LNs were identified by an expert pathologist as positive for cancer (P), and the remaining sixteen were negative (N). This database was divided into two groups. Group 1 (15P and 5N) was used for training and Group 2 (13P and 11N) was used for testing the classification technique. For all analysis each WSI was divided into non-overlapping 1000 x 1000 pixel sub-images that will be referred to as high-power fields (HPFs). For each LN in Group 1, at least one WSI was annotated by a pathologist to identify rectangular, HPF-scale regions as locally cancerous or locally non-cancerous. From these annotated slides, 924 HPFs (462 P and 462 N) were obtained. For each of these HPFs, statistical features based on gray-level co-occurrence matrices [1] and Law’s texture energy measures [2, 3] were extracted from 9 derived images [4]. The extracted features were submitted to a sequential forward selection (SFS) method [5] to select few non-redundant features providing best class separation (cancerous vs. non-cancerous region). Combinations of the selected features were tested on the 924 HPFs using k-fold cross-validation to find those that produced the best results and consequently to train our SVM-based classifier. In Group 2, WSI were not annotated for cancerous and non-cancerous zones on a HPF scale. Each LN, however, had been labeled by a pathologist as positive or negative for cancer. For each WSI, each section was divided into contiguous HPFs, and those which mainly contain fatty tissue, background, and tears were automatically excluded. Each selected HPFs was classified as cancerous or non-cancerous using the previously trained classifier to obtain the total number of cancer-classified per LN. A receiver operating characteristics (ROC) curve was traced by changing the discriminator threshold (T) used to label the LN as P for cancer as a function of the total number of cancer-classified HPFs. Results During training, 5 Laws features were selected by SFS. Highly satisfactory k-fold cross-validation with a F-score of 0.996 ± 0.005 was obtained using only 2 statistical features computed at different scales. The ROC curve obtained by applying the SVM-classifier to the test set is shown in the next figure. Two valuable operating points can be identified which both guaranteed no false-negative. At T=11 we got 2 false-positives and an optimal F-score of 0.917, and with a more conservative approach, T=1, we got 7 false-positives and a F-score of 0.759. The top-left part of the slide displayed in next figure would have been proposed to the pathologist as the most suspicious region of the cancerous LN.

A proteomic analysis of mushroom polysaccharide-treated HepG2 cells.

The anti-tumor properties of fungal polysaccharides have gained significant recognition in Asia and tropical America. In this study, the differential expression of proteins in normal HepG2 cells and those treated with polysaccharides that had been isolated from Phellinus linteus (PL), Ganoderma lucidum (GL) and Auricularia auricula (AA) was investigated. Using two-dimensional electrophoresis (2DE), a total of 104 protein spots were determined to be overexpressed in these cells compared with noncancerous regions. A total of 59 differentially expressed proteins were identified through MALDI-TOF-MS. In addition, 400 biological processes (BP), 133 cell components (CC) and 146 molecular functions (MF) were enriched by Gene Ontology (GO) analysis, and 78 KEGG pathways were enriched by pathway enrichment. Protein-Protein Interaction (PPI) analysis demonstrated the interaction networks affected by polysaccharides in HepG2 cells. Then, DJ-1 and 14-3-3 were identified as the key proteins in the networks, and the expression of the mRNA and proteins were evaluated using Real-time quantitative PCR (qRT-PCR) and Western blotting (WB), respectively. The results were in agreement with the 2DE. These results provided information on significant proteins of hepatocellular carcinoma (HCC) and form an important basis for the future development of valuable medicinal mushroom resources.

The use of matrix coating assisted by an electric field (MCAEF) to enhance mass spectrometric imaging of human prostate cancer biomarkers.

In this work, we combined a newly developed matrix coating technique - matrix coating assisted by an electric field (MCAEF) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to enhance the imaging of peptides and proteins in tissue specimens of human prostate cancer. MCAEF increased the signal-to-noise ratios of the detected proteins by a factor of 2 to 5, and 232 signals were detected within the m/z 3500-37500 mass range on a time-of-flight mass spectrometer and with the sinapinic acid MALDI matrix. Among these species, three proteins (S100-A9, S100-A10, and S100-A12) were only observed in the cancerous cell region and 14 proteins, including a fragment of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 2, a fragment of cAMP-regulated phosphoprotein 19, 3 apolipoproteins (C-I, A-I, and A-II), 2 S100 proteins (A6 and A8), β-microseminoprotein, tumor protein D52, α-1-acid glycoprotein 1, heat shock protein β-1, prostate-specific antigen, and 2 unidentified large peptides at m/z 5002.2 and 6704.2, showed significantly differential distributions at the p < 0.05 (t-test) level between the cancerous and the noncancerous regions of the tissue. Among these 17 species, the distributions of apolipoprotein C-I, S100-A6, and S100-A8 were verified by immunohistological staining. In summary, this study resulted in the imaging of the largest group of proteins in prostate cancer tissues by MALDI-MS reported thus far, and is the first to show a correlation between S100 proteins and prostate cancer in a MS imaging study. The successful imaging of the three proteins only found in the cancerous tissues, as well as those showing differential expressions demonstrated the potential of MCAEF-MALDI/MS for the in situ detection of potential cancer biomarkers. Copyright © 2015 John Wiley & Sons, Ltd.

COMBINING NONEXCHANGEABLE FUNCTIONAL OR SURVIVAL DATA SOURCES IN ONCOLOGY USING GENERALIZED MIXTURE COMMENSURATE PRIORS.

Conventional approaches to statistical inference preclude structures that facilitate incorporation of supplemental information acquired from similar circumstances. For example, the analysis of data obtained using perfusion computed tomography to characterize functional imaging biomarkers in cancerous regions of the liver can benefit from partially informative data collected concurrently in non-cancerous regions. This paper presents a hierarchical model structure that leverages all available information about a curve, using penalized splines, while accommodating important between-source features. Our proposed methods flexibly borrow strength from the supplemental data to a degree that reflects the commensurability of the supplemental curve with the primary curve. We investigate our method's properties for nonparametric regression via simulation, and apply it to a set of liver cancer data. We also apply our method for a semiparametric hazard model to data from a clinical trial that compares time to disease progression for three colorectal cancer treatments, while supplementing inference with information from a previous trial that tested the current standard of care.

Visualization of cancer-related chemical components in mouse pancreas tissue by tapping-mode scanning probe electrospray ionization mass spectrometry.

Mass spectrometry imaging is an informative approach for the comprehensive analysis of multiple components inside biological specimens. We used novel tapping-mode scanning probe electrospray ionization mass spectrometry method to visualize cancer-related chemical components in the mouse pancreas tissue section at a sampling pitch of 100 µm. Positive ion mode measurements from m/z 100 to 1500 resulted in the visualization of multiple components that are tentatively assigned as polyamines, lipids and proteins. Their signal intensities inside the cancerous and the non-cancerous regions were found to be significantly different by the two-sample t-test.

Association of MDR1 gene (C3435T) polymorphism and gene expression profiling in lung cancer patients treated with platinum-based chemotherapy.

Chemotherapy is the standard and recommended treatment for lung cancer apart from surgery and radiotherapy. Chemotherapy is administered as mono-agents or as combination therapy. In this study, we examined the role of MDR1 C3435T polymorphisms in lung cancer patients undergoing chemotherapy. We genotyped 126 cases with lung cancer and 111 healthy controls, using the polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). Frequencies of MDR1 C3435C, C3435T and T3435T genotypes were 61, 16 and 23% in lung cancer patients and 86, 9 and 5% in the controls, respectively. The T3435T genotypes had a 5.23-fold increased risk for lung cancer. (OR 5.23; 95% CI 2.082-13.146; p=0.0004). Patients with TT genotypes were more frequent in stage IV and were significantly associated with the disease (p=0.05). Habitual smoker lung cancer patients were 50% CC genotypes whereas TT genotypes were 34%. The non-smokers had 46% CC genotypes and 23% TT genotypes. Furthermore, we collected tissue biopsy samples for expression analysis from 20 patients (for controls we used the non-cancerous region of the same tissue). The present study showed mRNA expression of MDR1 was up-regulated in 80% of the cancer group in comparison with the control group (p=0.0002). We also correlated the association between MDR1 genotypes with different combinations of chemotherapy. The combinations and genotype distributions in the group receiving paclitaxel+cisplatin were as follows: CC (67%), CT (24%) and TT (9%) genotypes, respectively, and the group receiving carboplatin+gemcitabine CC (46%), CT (19%) and TT (35%) genotypes, respectively. We found that MDR1 (rs1045642) C3435T polymorphism and gene expression was significantly associated with the clinical outcome in lung carcinoma patients. In conclusion, it is suggested that MDR1 TT genotypes had higher risk for the development of lung cancer. Also, this polymorphism could be used as a genetic marker for predicting the clinical outcome of lung cancer patients.

Global loss of a nuclear lamina component, lamin A/C, and LINC complex components SUN1, SUN2, and nesprin-2 in breast cancer.

Cancer cells exhibit a variety of features indicative of atypical nuclei. However, the molecular mechanisms underlying these phenomena remain to be elucidated. The linker of nucleoskeleton and cytoskeleton (LINC) complex, a nuclear envelope protein complex consisting mainly of the SUN and nesprin proteins, connects nuclear lamina and cytoskeletal filaments and helps to regulate the size and shape of the nucleus. Using immunohistology, we found that a nuclear lamina component, lamin A/C and all of the investigated LINC complex components, SUN1, SUN2, and nesprin-2, were downregulated in human breast cancer tissues. In the majority of cases, we observed lower expression levels of these analytes in samples' cancerous regions as compared to their cancer-associated noncancerous regions (in cancerous regions, percentage of tissue samples exhibiting low protein expression: lamin A/C, 85% [n = 73]; SUN1, 88% [n = 43]; SUN2, 74% [n = 43]; and nesprin-2, 79% [n = 53]). Statistical analysis showed that the frequencies of recurrence and HER2 expression were negatively correlated with lamin A/C expression (P < 0.05), and intrinsic subtype and ki-67 level were associated with nesprin-2 expression (P < 0.05). In addition, combinatorial analysis using the above four parameters showed that all patients exhibited reduced expression of at least one of four components despite the tumor's pathological classification. Furthermore, several cultured breast cancer cell lines expressed less SUN1, SUN2, nesprin-2 mRNA, and lamin A/C compared to noncancerous mammary gland cells. Together, these results suggest that the strongly reduced expression of LINC complex and nuclear lamina components may play fundamental pathological functions in breast cancer progression.

Autophagy-based survival prognosis in human colorectal carcinoma.

The role of autophagy in cancers is controversial. Here we aim to determine the prognostic significance of autophagy in colorectal carcinoma patients, thereby allowing more rational development of therapeutic strategies. Through transmission electron microscopy, our data first demonstrated high frequency of defective mitochondria was strongly associated with poor overall survival in colorectal carcinoma. Next immunohistochemical study showed the expressions of Beclin 1, LC3B and Bcl-xL in both the center of tumor and adjacent noncancerous mucosal region were also correlated with overall survivals. We developed an autophagy signature for prognosis based on these three major autophagic proteins, further analysis suggested it was an independent prognostic biomarker and had its value even within single clinical stage. Combined TNM stage and this signature could significantly improve the accuracy of survival prognosis. To validate these immunohistochemical results, an internal testing cohort and an independent population were also included. Our findings suggest that autophagy plays an important role in the clinical cancer progression. Therefore autophagic proteins may be valuable prognostic biomarkers in the therapy of colorectal carcinoma and possibly other types of cancers.

Impact of Hepatic Steatosis on Disease-Free Survival in Patients with Non-B Non-C Hepatocellular Carcinoma Undergoing Hepatic Resection.

Although the prevalence of non-B non-C hepatocellular carcinoma (NBNC HCC) has increased, its clinicopathologic characteristics remain unclear. We retrospectively analyzed 518 HCC patients who underwent hepatic resection. Hepatitis B surface antigen- and hepatitis C antibody-negative patients were categorized into the NBNC HCC group (n = 145); others were categorized into the hepatitis B or C HCC (BC HCC) group (n = 373). We subdivided the etiologies of NBNC HCC according to alcohol intake and presence of steatosis. NBNC HCC was associated with nonalcoholic fatty liver disease (NAFLD) (13.1 %), fatty liver disease with moderate alcohol intake (9.0 %), alcoholic liver disease (ALD) (29.7 %), cryptogenic disease (44.1 %), and other known etiologies (4.1 %). The prevalence of obesity, diabetes mellitus, and hypertension was higher and hepatic function was better in the NBNC HCC group, which had significantly larger tumors than the BC HCC group. The entire NBNC HCC group displayed similar overall and disease-free survival as the BC HCC group. Among the subdivisions, NAFLD-associated HCC patients had significantly better disease-free survival than ALD-associated HCC and BC HCC patients. Microvascular invasion (hazard ratio [HR] 2.30; 95 % confidence interval [CI] 1.33-3.96) and steatosis area <5 % of noncancerous region (HR 2.13; 95 % CI 1.21-3.93) were associated with disease-free survival in NBNC HCC patients. The prognosis of NBNC HCC was similar to that of BC HCC. Among NBNC HCC patients, NAFLD-associated HCC patients had a relatively low recurrence risk. Absence of steatosis in hepatic parenchyma had a significant impact on disease-free survival in NBNC HCC patients.

Quantitative-ultrasound detection of cancer in human lymph nodes based on support vector machines

Histological assessment of lymph nodes excised from cancer patients suffers from an unsatisfactory rate of false-negative determinations. We are evaluating high-frequency quantitative ultrasound (QUS) to detect metastatic regions in lymph nodes freshly excised from cancer patients. Three-dimensional (3D) RF data were acquired from 289 lymph nodes of 82 colorectal-, 15 gastric-, and 70 breast-cancer patients with a custom scanner using a 26-MHz, single-element transducer. Following data acquisition, individual nodes underwent step-sectioning at 50-µm to assure that no clinically significant cancer foci were missed. RF datasets were analyzed using 3D regions-of-interest that were processed to yield 13 QUS estimates including spectral-based and envelope-statistics-based parameters. QUS estimates are associated with tissue microstructure and are hypothesized to provide contrast between non-cancerous and cancerous regions. Leave-one-out classifications, ROC curves, and areas under the ROC (AUC) were used to compare the performance of support vector machines (SVMs) and step-wise linear discriminant analyses (LDA). Results showed that SVM performance (AUC = 0.87) was superior to LDA performance (AUC = 0.78). These results suggest that QUS methods may provide an effective tool to guide pathologists towards suspicious regions and also indicate that classification accuracy can be improved using sophisticated and robust classification tools. [Supported in part by NIH grant CA100183.]

Different microRNA expression levels in gastric cancer depending on Helicobacter pylori infection.

Background/AimsThis study was conducted to identify microRNAs (miRNAs) that are differentially expressed in Helicobacter pylori-infected patients with an intestinal type of gastric cancer using miRNA microarray and to confirm the candidate miRNA expression levels.MethodsTotal RNA was extracted from the cancerous and noncancerous regions of formalin-fixed, paraffin-embedded tissues of H. pylori-positive (n=8) or H. pylori-negative (n=8) patients with an intestinal type of gastric cancer. RNA expression was analyzed using a 3,523 miRNA profiling microarray based on the Sanger miRBase. Validation analysis was performed using TaqMan miRNA assays.ResultsA total of 219 miRNAs in the aberrant miRNA profiles across the miRNA microarray showed at least a 2-fold change differential expression in H. pylori-positive and H. pylori-negative cancer tissues. After candidate miRNAs were selected using online miRNA databases, TaqMan miRNA assays confirmed that three miRNAs (miR-99b-3p, miR-564, and miR-638) were significantly increased in three H. pylori-positive cancer tissues compared to the H. pylori-negative cancer tissues. Additionally, four miRNAs (miR-204-5p, miR-338-5p, miR-375, and miR-548c-3p) were significantly increased in H. pylori-negative cancer tissues compared to H. pylori-positive cancer tissues.ConclusionsmiRNA expression in the intestinal type of H. pylori infection-dependent gastric cancer suggests that different gastric cancer pathogenesis mechanisms could exist between H. pylori-positive and H. pylori-negative gastric cancer. Additional functional studies are required.

Prostate cancer: diagnostic value of quantitative analysis by dynamic contrast-enhanced MR at 3.0 T

Objective To investigate the diagnostic value of the quantitative analysis parameters of dynamic contrast-enhanced MRI (DCE-MRI) for prostate cancer.Methods MR examination were performed in 26 patients and correlations were made with surgical pathology following radical prostatectomy.According to the localization of pathologic specimens,ROI were drawn on areas of cancerous regions,noncancerous regions in peripheral zone and central gland to measure the values of Ktrans,kep and Ve.The values of the three parameters in different tissue were compared with Kruskal-Wallis H and Mann-Whitney U.Results Twenty six patients had prostate cancer confirmed by pathology.Data from 468 assessable regions of prostate were acquired,including 115 cancerous regions and 142 noncancerous regions in central gland,68 cancerous regions and 143 noncancerous regions in peripheral zone.Prostate cancer showed a multifocal distribution.The K,kep and Ve values were (1.04 ± 0.26)/min,(1.43 ± 0.46)/min,(0.76 ±0.12) respectively for cancerous regions in central gland,and (0.82 ±0.19)/min,(1.12 ±0.26)/min,(0.75 ± 0.14) respectively for noncancerous regions in central gland,(0.95 ±0.31)/min,(1.31 ±0.51)/min,(0.76 ± 0.13) respectively for cancerous regions in peripheral zone and (0.32 ± 0.07)/min,(0.52 ± 0.13)/min,(0.64 ± 0.14) respectively for noncancerous regions in peripheral zone.The differences among the three parameters were statistically significant (x2 =316.008,297.985,63.681,P ＜0.01).The Ktrans,kep values of cancerous regions were significantly higher than the corresponding values of noncancerous regions,respectively.Conclusion Quantitative analysis parameters of DCE-MRI contribute to the diagnosis of prostate cancer,including cancer located in central gland. Key words: Prostate neoplasms; Magnetic resonance imaging; Pathology

KL-6 mucin expression in pancreatic ductal carcinoma and its clinical significance

Objective To investigate the expression of KL-6 mucin in pancreatic ducatal tissue,and to explore its clinical significance.Methods All specimens form 38 patients who were diagnosed as pancreatic ductal adenocarcinoma were subjected to immunohistochemical analysis using KL-6 monoclonal antibody.Results 100％ specimens of pancreatic adenocarcinoma were found positive staining,and marked positive staining was 78.9％,remarkable positive staining was not found in the surrounding noncancer regions of the pancreatic tissues (72.3％).KL-6 mucin expression was statistical significance with the tumor development and differentiation.Conclusion Most of all pancreatic adenocarcinoma cases showed remarkable positive staining for KL-6 mucin,while the majority of surrounding non-cancer regions did not.The aberrant expression of KL-6 mucin is significantly related to aggressive tumor behaviors of pancreatic adenocarcinoma.KL-6 mucin may be a specific target to detect the pancreatic adenocarcinoma in molecular imaging study. Key words: Pancreatic ductal adenocarcinoma; KL-6 mucin; Immunohistochemical

P28GANK is a novel marker for prognosis and therapeutic target in gastric cancer

P28/PSMD10, a regulatory complex of the human 26S proteasome, plays a critical role in tumor genesis. This study was designed to clarify the clinical significance of p28GANK in gastric cancer. In order to demonstrate the importance of p28GANK expression for the prognosis of gastric cancer, p28GANK expression in 124 paired cases of gastric cancer and noncancerous regions and immortal gastric epithelial cell GES-1 and 5 human gastric cancer cell lines was analyzed by RT-PCR in real-time. MTT was used to observe the effect of P28GANK on cell growth. P28GANK expression was higher in gastric cancer tissues than in corresponding normal tissues (p = 0.0033), and patients comprising the group with p28GANK high expression had a poorer overall survival rate than those from the low expression group (p = 0.0037). Further, the results of multivariate analysis demonstrated that the high p28GANK expression was an independent prognostic factor of gastric cancer process. P28GANK expression was also up-regulated in five gastric cancer cell lines. As it has been shown by in vitro proliferation assay, p28GANK expression correlated with tumor growth. On the base results of present study one can suggests the p28GANK being useful as a predictive marker for patient prognosis and a novel therapeutic target for gastric cancer.

The significance of LRPPRC overexpression in gastric cancer

LRPPRC is a multifunctional protein involved in mitochondrial gene expression and function, cell cycle progression, and tumorigenesis. We analyzed LRPPRC gene expression in 253 paired cases of gastric cancer and noncancerous regions and six gastric cancer cell lines to demonstrate the importance of LRPPRC expression for the prediction of prognosis of gastric cancer. Our results showed that LRPPRC expression in gastric cancer tissues is significantly higher than that in paired control tissue (P < 0.001). Patients with higher LRPPRC expression showed a poorer overall survival rate than those with lower LRPPRC expression (P < 0.001). Multivariate analysis demonstrated that lymph node metastasis (N), distant metastasis (M), TNM stage, and LRPPRC expression were independent prognostic factors for gastric cancer (P = 0.004, 0.002, 0.017, 0.004 respectively).Moreover, Western blotting showed that LRPPRC expression was increased in SGC7901, BGC823, MKN45, and XGC9811cells. The in vitro proliferation assay showed that LRPPRC expression is inversely associated with gastric cancer cells growth. Our results indicated that LRPPRC could be used as a predictive marker for patient prognosis of gastric cancer and may be a novel therapeutic target for gastric cancer in future.

Stem cell self-renewal factors Bmi1 and HMGA2 in head and neck squamous cell carcinoma: clues for diagnosis

Head and neck squamous cell carcinoma (HNSCC) includes both morphological and functional cellular heterogeneity, as would be expected if it arose from dysregulated stem or progenitor cells as opposed to the simple clonal expansion of a mutated cell; however, stemness molecule expression levels and distribution in HNSCC remain unclear. To clarify this, stemness molecule expressions were determined in HNSCC, as well as their properties and prognosis. Two proto-oncogenic chromatin regulators, Bmi-1 and high-mobility-group A2 (Hmga2), were identified in 12 pair cases of HNSCC tumor regions by comparison with their non-cancerous background tissues using cDNA microarray. Both Bmi-1 and Hmga2 are known to promote stem cell self-renewal by negatively regulating the expressions of Ink4a and Arf tumor suppressors. Despite similar targets, Bmi-1 protein was expressed in an early cancerous region and HMGA2 protein was expressed in a region showing more progression. Similarly, Bmi1 expression had no significance with regard to overall survival (P=0.67), whereas HMGA2 expression was associated with decreased overall survival (P=0.05). Quantitative real-time reverse transcription polymerase chain reaction analyses also correlated with protein levels. These findings suggest that Bmi-1 is an early detection marker to distinguish cancerous from non-cancerous regions, whereas HMGA2 is presumed to be a tumor prognosis marker. Among our HNSCC analyses, these stemness molecules expressed fewer primitive rare cells in the tumor than all other cells in the tumor. HNSCC cells with high expression of stemness molecules partly behave like stem cells.