Strong Cytoplasmic Expression Research Articles

Endoplasmic Reticulum Stress in UMOD-Related Kidney Disease: A Human Pathologic Study

Mutations of the UMOD gene, which encodes the uromodulin protein, are associated with tubulointerstitial nephritis and hyperuricemia. UMOD mutations impair uromodulin folding, resulting in its retention within the endoplasmic reticulum (ER) of renal tubular cells. The aim of this study was to investigate whether mutant uromodulin accumulation in epithelial tubular cells is associated with ER stress. We characterized tubular expression of uromodulin and the ER stress surrogate marker Grp78 by immunohistochemistry in kidney biopsy specimens from 7 patients with UMOD-related kidney disease. We compared this population with 5 patients with familial tubulointerstitial nephritis not related to UMOD mutation. All biopsy specimens from patients carrying the UMOD mutation showed strong heterogeneous cytoplasmic expression of uromodulin in cells of the thick ascending limb of the loop of Henle. In the same tubules, Grp78 was highly expressed in a perinuclear pattern. In contrast, in all kidney biopsy specimens from patients without UMOD mutations, uromodulin staining showed normal apical expression and Grp78 expression was not increased. Our observations support the hypothesis that ER accumulation of mutant uromodulin may cause ER stress, providing a potential mechanism for the progression of UMOD-related kidney disease.

Expression of Cytokeratin 10 Protein in the Human Testis Showing Normal and Abnormal Spermatogenesis

Background: CK10 is a heterotetramer of type I and two type II keratins. Aim: This study examines the expression pattern of cytokeratin 10 (CK10) in human testis. Materials and methods: CK10 protein expression was examined using immunofluorescense staining methods in 30 human testicular biopsy specimens (normal spermatogenesis, maturation arrest and Sertoli cell only syndrome, 10 cases each) obtained from patients undergoing investigations for infertility. Results: In the testis showing normal spermatogenesis, CK10 was expressed in the interstitium and in the seminiferous tubules. A strong cytoplasmic expression was seen in the Leydig cells, Sertoli cells, and spermatocytes. In testes showing spermatogenic arrest, weak CK10 protein expression was observed both in the interstitium and seminiferous tubules (some primary spermatocytes). In the testes showing Sertoli cell only syndrome, negligible CK10 staining was seen both in the seminiferous tubules and in the interstitial cells of Leydig. Conclusions: To the authors’ knowledge, this is the first study indicating CK10 expression in the human testis during normal and abnormal spermatogenesis. The varied expression of CK10 in testes showing abnormal spermatogenesis suggests its possible involvement in this process.

Nuclear and cytoplasmic LIMK1 enhances human breast cancer progression

BackgroundLIM kinase 1 (LIMK1) is expressed in both cytoplasmic and nuclear compartments, and is a key regulator of cytoskeletal organization involved in cell migration and proliferation. LIMK1 levels are increased in several human cancers, with LIMK1 over-expression in prostate and breast cancer cells leading to tumor progression. While it has been presumed that the mechanism by which LIMK1 promotes cancer progression is via its cytoplasmic effects, the role of nuclear vs cytoplasmic LIMK1 in the tumorigenic process has not been examined.ResultsTo determine if cytoplasmic or nuclear LIMK1 expression correlated with breast cancer, we performed immunohistochemical (IHC) analysis of breast tissue microarrays (TMAs), The IHC analysis of breast TMAs revealed that 76% of malignant breast tissue samples strongly expressed LIMK1 in the cytoplasm, with 52% of these specimens also expressing nuclear LIMK1. Only 48% of benign breast samples displayed strong cytoplasmic LIMK1 expression and 27% of these expressed nuclear LIMK1. To investigate the respective roles of cytoplamsic and nuclear LIMK1 in breast cancer progression, we targeted GFP-LIMK1 to cytoplasmic and nuclear subcellular compartments by fusing nuclear export signals (NESs) or nuclear localization sequences (NLS), respectively, to the amino-terminus of GFP-LIMK1. Stable pools of MDA-MB-231 cells were generated by retroviral transduction, and fluorescence microscopy revealed that GFP alone (control) and GFP-LIMK1 were each expressed in both the cytoplasm and nucleus of MDA-MB-231 cells, whereas NLS-GFP-LIMK1 was expressed in the nucleus and NES-GFP-LIMK1 was expressed in the cytoplasm. Western blot analyses revealed equal expression of GFP-LIMK1 and NES-GFP-LIMK1, with NLS-GFP-LIMK1 expression being less but equal to endogenous LIMK1. Also, Western blotting revealed increased levels of phospho-cofilin, phospho-FAK, phospho-paxillin, phospho-Src, phospho-AKT, and phospho-Erk1/2 in cells expressing all GFP-LIMK1 fusions, compared to GFP alone. Invasion assays revealed that all GFP-LIMK1 fusions increased MDA-MB-231 cell invasion ~1.5-fold, compared to GFP-only control cells. Tumor xenograft studies in nude mice revealed that MDA-MB-231 cells stably expressing GFP-LIMK, NLS-GFP-LIMK1 and NES-GFP-LIMK1 enhanced tumor growth 2.5-, 1.6- and 4.7-fold, respectively, compared to GFP-alone.ConclusionTaken together, these data demonstrate that LIMK1 activity in both the cytoplasmic and nuclear compartments promotes breast cancer progression, underscoring that nuclear LIMK1 contributes to the transforming function of LIMK1.

Anaplastic lymphoma kinase (ALK) in rhabdomyosarcoma.

9530 Background: Rhabdomyosarcoma (RMS) is a highly malignant soft tissue sarcoma that occurs predominantly in children. Despite the use of multi-agent chemotherapy, survival for patients with alveolar histology (ARMS), metastatic and recurrent disease remains poor. Moreover, the intensive treatment regimens result in a high rate of adverse effects. This urges the development of new therapeutic strategies in RMS. ALK inhibitors are currently available for clinical trials. The aim of this study is to investigate ALK as a potential therapeutic target in RMS, with emphasis on protein expression and the underlying genomic aberrations. Methods: Paraffin-embedded RMS tissue samples were collected on tissue microarrays. ALK protein expression was evaluated by immunohistochemistry (ALK1, DAKO). A semiquantitative scoring system was used in which staining was scored negative “0”, and weak “1” or strong “2” when present in at least 10% of the tumor cells. ALK gene copy number and presence of ALK translocation were determined by in situ hybridization. Copy number was divided into normal (0-4 copies), gain (5-10 copies), or amplification (>10 copies) with a cut-off at 10% of the nuclei. LAF (2q11) was used as a reference probe for chromosome 2 aberrations. Results: A total of 107 primary RMS tumors were evaluated (83 embryonal (ERMS) and 24 ARMS). Strong cytoplasmic ALK protein expression was observed more frequently in ARMS than in ERMS (90.9% versus 35.8%, p<0.001). ALK gene copy number gain was detected in the vast majority of ARMS (overall 59.6%; ARMS 90.0%, ERMS 52.1%, p=0.002), and one amplification was identified (ARMS). A significant correlation was observed between ALK copy number gain and strong ALK protein expression (p<0.001). In ERMS cases with ALK gain evaluable for LAF copy number analysis (N=31), ALK/LAF ratio (range 1.10-2.72) was 1.5-2 in 54.8%, and >2 in 12.9%. In ARMS (N=16), ALK/LAF ratio (range 0.95-2.18) was 1.5-2 in 25%, and >2 in 18,75%. No ALK translocations were identified. Conclusions: This study reveals that ALK is a promising target for future treatment of RMS patients. Copy number gains appeared to be one of the underlying mechanisms for ALK protein expression. Further insight into additional genetic aberrations should be subject of future research.

Malignant mesothelioma of the peritoneum invading the liver and mimicking metastatic carcinoma: A case report

We present a case of malignant mesothelioma of the peritoneum with massive direct invasion to the liver in a 58-year-old Japanese woman. She had no history of asbestos exposure or other malignancies. Abdominal computed tomography revealed one 8-cm intrahepatic mass adjacent to the abdominal wall with peritoneal thickening, multiple smaller nodules in the peritoneal cavity, and intra-abdominal lymphadenopathy. Liver biopsy showed a small cluster of atypical cells similar to epithelial neoplasm, which formed a tubulopapillary structure. The tumor cells were positive for calretinin with strong nuclear and cytoplasmic expression together with podoplanin (D2-40) and some cytokeratins, but were negative for hepatocyte paraffin 1 and other adenocarcinoma markers. We confirmed a diffuse peritoneal mesothelioma with direct invasion to the liver. Liver masses with other peritoneal nodules are mostly encountered as metastatic diseases. However, the possibility of mesothelioma should be considered, even in women without an apparent history of asbestos exposure.

P16 Expression: A marker of differentiation between childhood malignant melanomas and Spitz nevi

Childhood malignant melanomas frequently present as nodular melanomas with Spitzoid features. Spitz nevus and Spitzoid melanoma overlap clinically and histopathologically and there have been many attempts to differentiate between them. Spitz nevi differ from melanomas by their immunohistochemical pattern of expression of cell cycle and apoptosis regulators such as the p16 protein. The aim of this study was to evaluate in a childhood population the expression of p16 in nodular malignant melanoma of Spitzoid type, Spitz nevi, and a control group of benign compound melanocytic nevi. We performed immunohistochemical studies for expression of p16 in 6 Spitzoid malignant melanomas, 18 Spitz nevi, and 12 compound melanocytic nevi in children younger than 18 years. Statistical analysis was used to compare p16 expression, mitotic count/mm(2), and Ki-67 index of childhood nodular malignant melanomas and Spitz nevi. All the childhood melanoma cases were associated with loss of p16 without any correlation with their Breslow thickness whereas all the Spitz nevi and benign melanocytic nevi had strong positive nuclear and cytoplasmic expression of p16 staining. We found a statistically significant difference in p16 expression, mitotic counts, and Ki-67 index when comparing the Spitzoid melanomas with the Spitz nevi. This study is limited by the small number of malignant melanomas, which are known to be rare in childhood. p16 Expression in childhood nodular Spitzoid malignant melanomas and Spitz nevi, in conjunction with clinical and histopathological evaluation, may be a useful tool in differentiating between these two entities.

Abstract 300: Identification of a panel of protein markers associated with disease progression in bladder cancer

Abstract Most bladder cancer patients are initially diagnosed with noninvasive or superficially invasive urothelial tumors, comprising benign low grade Ta tumors that rarely progress to high grade T1 tumors with concomitant CIS that progress in up to 60% of cases. So far, no biomarkers are available for solid prediction of the individual disease course. Previously, we identified an 88-transcript signature for prediction of progression for patients with non-muscle invasive bladder cancer. Aim: validation of key candidate markers on the protein level. We selected four protein coding genes, where high transcript levels were related to a reduced risk of progression and five genes where high expression identified patients with an increased risk of progression. Microarray transcript profiling data on 7 bladder cancer cell lines were used to select appropriate model cell lines. Specificity of antibodies against the selected proteins, protein expression and potential splice variants were investigated by western blotting using extracts from bladder cancer cell lines (T24, SW780) and HeLa cells. Specific antibodies were identified for six of the nine selected proteins. Immunohistochemistry was applied to a Tissue-microarray with 289 primary non-muscle invasive urothelial tumors (182 pTa, 101 pT1 and 6 CIS). 118 patients later experienced disease progression to pT2-4 bladder cancer (median time to progression 22 months). None of the remaining 171 patients showed later disease progression within the follow-up period (median follow-up 93 months). In univariate Cox regression analysis, expression of 4 of the 6 proteins correlated significantly with disease progression from stage Ta or T1 to muscle invasive bladder cancer. A reduced risk of progression correlated significantly with a strong nuclear expression of the G1/S-specific cell cycle protein (p=0.0003) and its expression in &amp;gt;80% nuclei (p=0.0004) as well as a membranous localization of a disintegrin and metallopeptidase (p = 0.0002). Increased risk of progression correlated significantly with strong nuclear and cytoplasmic expression of an ubiquitin-conjugating enzyme (p=0.0001) and its expression in &amp;gt;10% nuclei (p=0.0000) and with the expression of a DNA replication licensing factor in &amp;gt;80% nuclei (p=0.0026). In multivariate Cox regression analysis, the G1/S-specific cell cycle protein (HR=0.62, p=0.020, 95%CI 0.42-0.92), the disintegrin and metallopeptidase (HR=0.56, p=0.008, 95%CI 0.37-0.86) and the ubiquitin-conjugating enzyme (HR=1.68, p=0.015, 95%CI 1.1-2.5) were independent prognostic variables for predicting disease progression to muscle invasive bladder cancer when stratifying for other known clinical and histopathological risk factors as stage, age and grade. The Ubiquitin-conjugating enzyme was selected for in vitro functional analyses probably affecting cell proliferation and cell migration of bladder cancer cell lines. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 300. doi:10.1158/1538-7445.AM2011-300

Targeting p21-activated kinase 1 (PAK1) to induce apoptosis of tumor cells

p21-activated kinases (PAKs) are serine/threonine protein kinases that serve as important mediators of Rac and Cdc42 GTPase function as well as pathways required for Ras-driven tumorigenesis. PAK1 has been implicated in signaling by growth factor receptors and morphogenetic processes that control cell polarity, invasion, and actin cytoskeleton organization. To better understand the role of PAK1 in tumorigenesis, PAK1 genomic copy number and expression were determined for a large panel of breast, lung, and head and neck tumors. PAK1 genomic amplification at 11q13 was prevalent in luminal breast cancer, and PAK1 protein expression was associated with lymph node metastasis. Breast cancer cells with PAK1 genomic amplification rapidly underwent apoptosis after inhibition of this kinase. Strong nuclear and cytoplasmic PAK1 expression was also prevalent in squamous nonsmall cell lung carcinomas (NSCLCs), and selective PAK1 inhibition was associated with delayed cell-cycle progression in vitro and in vivo. NSCLC cells were profiled using a library of pathway-targeted small-molecule inhibitors, and several synergistic combination therapies, including combination with antagonists of inhibitor of apoptosis proteins, were revealed for PAK1. Dual inhibition of PAK1 and X chromosome-linked inhibitor of apoptosis efficiently increased effector caspase activation and apoptosis of NSCLC cells. Together, our results provide evidence for dysregulation of PAK1 in breast and squamous NSCLCs and a role for PAK1 in cellular survival and proliferation in these indications.

Expression of CD1d Protein in Human Testis Showing Normal and Abnormal Spermatogenesis

CD1d is a member of CD1 family of transmembrane glycoproteins, which represent antigen-presenting molecules. Immunofluorescent staining methods were utilized to examine expression pattern of CD1d in human testicular specimens. In testis showing normal spermatogenesis, a strong CD1d cytoplasmic expression was seen the Sertoli cells, spermatogonia, and Leydig cells. A moderate expression was observed in the spermatocytes. In testes showing maturation arrest, CD1d expression was strong in the Sertoli cells and weak in spermatogonia and spermatocytes compared to testis with normal spermatogenesis. In Sertoli cell only syndrome, CD1d expression was strong in the Sertoli and Leydig cells. This preliminary study displayed testicular infertility-related changes in CD1d expression. The ultrastructural changes associated with with normal and abnormal spermatogenesis are open for further investigations.

Spontaneous Regression of Highly Immunogenic Molluscum contagiosum Virus (MCV)-Induced Skin Lesions Is Associated with Plasmacytoid Dendritic Cells and IFN-DC Infiltration

Molluscum contagiosum virus (MCV) infection induces self-limiting cutaneous lesions in an immunocompetent host that can undergo spontaneous regression preceded by local inflammation. On histology, a large majority of MCV-induced lesions are characterized by islands of hyperplastic epithelium containing infected keratinocytes and surrounded by scarce inflammatory infiltrate. However, spontaneous regression has been associated with the occurrence of a dense inflammatory reaction. By histology and immunohistochemistry, we identified MCV-induced lesions showing a dense inflammatory infiltrate associated with cell death in keratinocytes (inflammatory Molluscum contagiosum (I-MC)). In I-MC, hyperplastic keratinocytes were highly immunogenic as demonstrated by the expression of major histocompatibility complex class I and II molecules. Immune cell infiltration consisted of numerous cytotoxic T cells admixed with natural killer cells and plasmacytoid dendritic cells (PDCs). Accordingly, a type I IFN signature associated with PDC infiltration was demonstrated in both keratinocytes and inflammatory cells. Among the latter, a cell population resembling IFN-DC (CD123(+)CD11c(+)CD16(+)CD14(+)MxA(+)) was identified in proximity to islands of apoptotic keratinocytes. In vitro-generated IFN-DCs expressed a strong cytotoxic signature, as demonstrated by high levels of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas ligand (FasL). This study establishes a previously unreported model to underpin the role of innate immune cells in viral immune surveillance.

Androgen receptor coactivator p44/Mep50 in breast cancer growth and invasion

Hormones and their receptors play an important role in the development and progression of breast carcinoma. Although the primary focus has been on oestrogen and oestrogen receptor (ER), androgen, androgen receptor (AR) and its coactivator(s) have been implicated in tumorigenesis of breast carcinoma and warrant further investigation. AR coactivator p44/Mep50 is identified as a subunit of methylosome complex and lately characterized as an AR coactivator that enhances AR mediated transcription activity in a ligand dependent manner. In prostate cancer, p44 is expressed in the nucleus of benign epithelia and translocated into the cytoplasm in cancer cells. Furthermore, nuclear expression of p44 inhibits prostate cancer growth. In this report, we examined the expression and function of p44 in breast cancer. In addition to being an AR coactivator, p44 also functions as an ER coactivator. In contrast to findings in prostate cancer, the expression of p44 shows strong cytoplasmic expression in morphologically normal terminal ductal lobular units, while nuclear p44 is observed in both ductal carcinoma in situ and invasive carcinoma. Further, overexpression of nuclear-localized p44 stimulates proliferation and invasion in MCF7 breast cancer cells in the presence of oestrogen and the process is ERα dependent. These findings strongly suggest that p44 plays a role in mediating the effects of hormones during tumorigenesis in breast.

Strong cytoplasmic expression of COX2 at the invasive fronts of gallbladder cancer is associated with a poor prognosis

AimThe significance of cell cycle proteins, p21 and p53, and cyclo-oxygenase-2 (COX2) is still controversial in gallbladder adenocarcinoma. Conflicting results exist with regard to the prognostic significance of p21 and...

Aberrant expression of the pluripotency marker SOX-2 in endometriosis

Expression of the pluripotency factors SOX-2, OCT-4, KLF-4, and NANOG was analyzed by quantitative real-time polymerase chain reaction, immunohistochemistry, and immunofluorescence microscopy in the endometrium, myometrium, and endometriotic tissue of 36 patients. Aberrant expression of SOX-2 may indicate a stem cell origin of endometriosis, whereas the presence of all progenitor markers in endometrial tissue marks the endometrium as a potential source for induced pluripotent stem cell generation.

P16 Expression in Squamous Cell Carcinomas of Cervix and Bladder

p16 is a widely used immunohistochemical marker in gynecologic pathology. Strong and diffuse cytoplasmic and nuclear expression of p16 in squamous cell carcinomas (SCC) of the female genital tract is strongly associated with high-risk human papilloma virus infection and neoplasms of cervical origin. However, p16 can be expressed in other neoplasms and in several normal human tissues. Occasionally, SCCs may involve both uterine cervix and urinary bladder. Accurate identification of the site of origin in such cases has therapeutic and prognostic implications. We investigate the potential value of p16 expression in this distinction. We reviewed 74 SCCs, 38 (51%) from urinary bladder and 36 (49%) from uterine cervix obtained between 2003 and 2008. Of the 38 cases of bladder carcinoma, 21 occurred in females and 17 in males. Immunohistochemical analysis for p16 (DAKO M7247, clone 484, dilution of 1:50) expression was done in all cases using the labeled streptavidin-biotin method. Strong and diffuse nuclear and cytoplasmic p16 positivity was observed in 45 cases (61%). Of the 38 SCCs of urinary bladder, 14 (37%) expressed p16 (8 males, 6 females). Of the 36 SCCs of uterine cervix, 31 (86%) were positive for p16. (1) The majority of SCCs of uterine cervix express p16. (2) More than a third of urinary bladder SCCs express p16. (3) SCCs of urinary bladder express p16 independent of gender. (4) p16 immunohistochemical expression alone cannot be used to discriminate between SCCs arising from uterine cervix versus urinary bladder.

Abstract 2241: Role of tissue markers in the prediction of bone metastases from breast cancer

Abstract Introduction: Bone metastasis is a frequent event in patients with breast cancer. OPG (osteoprotegerin) protects bone from excessive resorption by binding to RANK-L (receptor activator of nuclear factor-kB-ligand) and by preventing it from binding to RANK. These molecules are closely involved in the process of metastasization to bone. CXCL12/SDF-1α, the ligand of CXCR4, is overexpressed in bone, and the local chemokine milieu is now emerging as a key determinant in organ selectivity by tumor cells. We carried out a retrospective analysis to evaluate the potential role of these biological markers in predicting bone metastasization in breast cancer patients. Materials and methods: OPG and CXCR4 expression was determined by avidin-biotin immunohistochemistry using a polyclonal (H-249, Santa Cruz) and monoclonal (Ab58176, Abcam) antibody, respectively. Immunohistochemical staining was carried out on sections from paraffin-embedded blocks of 40 primary breast cancers. Ten patients (median age of 64 years, range 48-78) were disease-free (DF) at a median of 9.8 (range 6.9-11.5) years, while 30 (median age 67 years, range 42-87) had relapsed. In the latter group, 10 (median age 66 years, range 42-87) had visceral metastases (VM) and 20 (median age 69 years, range 42-87) had bone metastases (BM). Results: Considering only strong cytoplasmic expression as positive, 22.5% of tumors were positive for OPG and 25% for CXCR4. In particular, OPG was expressed in 20% of tumors in DF patients and in 25% in BM patients. Conversely, CXCR4 was expressed in 10% of tumors in DF patients and in 45% in BM patients. OPG was expressed in only 20% of tumors in VM patients. There was no correlation between OPG and CXCR4 expression, and positivity to at least one of the two markers was observed in 30% of tumors in DF patients, 20% in VM patients and 55% in BM patients. In this last subgroup, there were no statistically significant associations between marker expression and tumor characteristics. Furthermore, no relation was observed between marker expression and disease-free or overall survival in BM patients. Conclusions: Our preliminary results suggest that cytoplasmic CXCR4 expression in primary breast cancers could play a role in predicting bone metastases. Enrolment of breast cancer patients in a larger study is ongoing to confirm these data and to evaluate the predictive role of RANK and RANK-L in bone metastasization. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2241.

Abstract 1729: Cytoplasmic expression of survivin is an independent predictor of poor prognosis in patients with salivary gland cancer

Abstract Background: The expression of the inhibitor of apoptosis protein survivin is shown to be a significant prognostic indicator in various human cancers. Its role in cancer of the major salivary glands remains unclear. Methods: We analysed the survivin expression in 171 patients with malignant parotid gland cancer (12 different histological subtypes) by means of immunohistochemistry. The experimental findings were correlated with clinicopathological and survival parameters. The mean follow-up time was 42 months. Results: There was no significant correlation of cytoplasmic survivin expression levels with the patients’ clinicopathological characteristics. A strong cytoplasmic expression of survivin was found in 25% of the examined tumors. High cytoplasmic survivin expression was an independent prognostic indicator for a significant poorer 5-year disease-free survival rate (31% versus 67%, p&amp;lt;0.0005) and overall survival (45% versus 62%, p=0.048). In the multivariate analysis survivn expression and distant metastasis were significant predictors for patient outcome (p=0.001, p&amp;lt;0.0005). Conclusions: The correlation between cytoplasmic survivin expression and survival probabilities of salivary gland malignancies might make this an effective tool in patient follow-up, prognosis and targeted therapy in future. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1729.

Macrophage chemotactic protein-1 mRNA levels in non-Hodgkin lymphoma

Non-Hodgkin lymphoma (NHL) is one of the most common malignancies whose incidence increases, and the treatment results are not satisfactory. The aim of this study was to determine the capacity of NHL to produce MCP-1, chemokine that induces chemotaxis of macrophages and lymphoid cells. The mRNA expression and protein MCP-1 expression were determined in the samples of 20 patients with NHL and 8 reactive tonsils. MCP-1 mRNA was detected in 8/8 tonsils and in 19/20 patients with NHL by real-time PCR analysis. In addition, the amount of detected MCP-1 cDNA was significantly higher in patients with limited stage, good IPI, normal level of fibrinogen and LDH. Finally, in patients with aggressive NHL, the level of MCP-1 cDNA was higher than in indolent tumours. Immunohistochemical analysis revealed that majority of stromal elements such as macrophages, endothelial and smooth muscle cells in reactive as well as in neoplastic lymphoid tissue showed strong cytoplasmic MCP-1 expression. Moderate cytoplasmic MCP-1 expression was also observed in reactive lymphocytes, while tumour cells of indolent NHL were mostly pale in comparison with aggressive lymphomas which predominantly demonstrated intense MCP-1 staining. These intriguing preliminary results emphasize the need for further investigations that must be conducted on the representative sample with concordant measurement of serum MCP-1 level.

Expression of p16INK4A in gastrointestinal stromal tumours (GISTs): two different forms exist that independently correlate with poor prognosis

To determine the prognostic impact of p16INK4A expression in gastrointestinal stromal tumours (GISTs), which is currently being questioned, with both loss and overexpression said to be correlated with poor prognosis. Two different forms of p16INK4A were identified, presenting with predominantly nuclear and cytoplasmic expression pattern, respectively. The immunohistochemical expression of the two forms and their correlation with E2F1 and prognosis were analysed in a series of 120 GISTs with clinical follow-up. Low nuclear p16INK4A expression correlated with E2F1 up-regulation, higher mitotic counts, and tumour progression. The prognostic value of nuclear p16INK4A expression was only marginally significant (P=0.05). Strong expression of the cytoplasmic p16INK4A form was significantly associated with shorter disease-free survival (P=2x10(-5)). The prognostic impact of strong expression of the cytoplasmic p16INK4A form was independent of anatomical localization, tumour size and mitotic counts, and significant even among the cohort of tumours with high malignant potential. Low expression of the nuclear p16INK4A form and strong expression of the cytoplasmic p16INK4A form both represent two independent parameters each associated with tumour progression in GISTs. Low nuclear p16INK4A expression enables E2F1 up-regulation and consecutive accelerated cell proliferation. In contrast, strong cytoplasmic p16INK4A expression probably reflects a negative feedback loop as a result of (as yet unknown) oncogenic events.

Nuclear surviving expression predicts poor prognosis in cholangiocarcinonm

Objective To study the prognostic role of survivin expressed in cholangiocarcinoma. Methods Forty-eight cases of cholangiocarcinoma were studied. Immunohistochemistry was performed u-sing a monoclonal antibody to survivin. Survivin expression was described as absent or weak and strong. Results Cytoplasmic and nuclear survivin expression was seen in 26 and 22 patients respectively. Strong cytoplasmic survivin expression was seen in 12 cases and strong nuclear survivin in 8. Patients with strong nuclear survivin had a median survival of 10 months, significantly lower than for patients with weak nuclear survivin expression (19months,P ＜0.05). Multivariate analysis using the Cox-propertional hazards model identified 3 important prognostic predictors: nuclear survivin (P ＜ 0.05) and presence of metastasis (P ＜0.05). Conclusion Nuclear survivin expression in cholangiocarcinoma may predict a poor prognosis. Key words: Survivin; Cholangiocarcinoma; Immunohistochemistry

A case of complete androgen insensitivity syndrome presenting with incarcerated inguinal hernia: an immunohistochemical study

To present a case of complete androgen insensitivity syndrome that initially presented as incarcerated inguinal hernia and to investigate the expression of antimüllerian hormone (AMH), SOX-9 (SRY-box-containing gene 9), prostaglandin D synthase (PGDS), and androgen receptors (AR). Case report. District hospital. A 12-year-old girl with negative past medical and family history, who presented with a 6-hour history of progressive left groin pain. Open gonadectomy. Immediate surgery, diagnosis, and referral to a multidisciplinary team for further management. Pathologic analysis revealed a left twisted and infarcted testicle, and a normal right one containing immature seminiferous tubules with a decreased number of spermatogonia. Immunohistochemical analysis revealed strong cytoplasmic and nuclear expression of AMH and SOX-9, respectively, by the Sertoli cells as well as focal weak cytoplasmic PGDS expression. The spermatogonia showed focal weak nuclear PGDS expression. The Leydig cells showed no immunoreactivity at all. No AR immunoreactivity was observed. Negative AR immunostaining could either reflect Sertoli-cell immaturity or a mutation resulting in no protein production. The AMH immunodetection, consistent with its high serum levels, could potentially reflect Sertoli-cell immaturity. The SOX-9 nuclear detection in the Sertoli cells was consistent with its role, inducing male sex differentiation, including AMH expression. The nuclear localization of PGDS in the spermatogonia needs further investigation.