P53 Immunoreactivity Research Articles

Dysplasia Involves Enteritis Cystica Profunda in the Setting of Crohn’s Disease

Enteritis cystica profunda is a rare non-neoplastic condition characterized by misplaced glands deep to the muscularis mucosae of the small bowel, likely as a result of trauma or inflammation such as Crohn’s disease, a disease with increased risk for small bowel adenocarcinoma. Clinically, enteric cystica profunda and adenocarcinoma can present with obstruction and/or stricture in patients with Crohn’s disease and should be differentiated from each other. Neoplastic involvement of enteritis cystica profunda by dysplasia has not been reported previously. Herein, we report a case of enteritis cystica profunda involved by high-grade dysplasia of gastric type. High-grade dysplasia and intramucosal adenocarcinoma of gastric type were also present in the overlying mucosa. Additionally, the adjacent mucosa demonstrated marked chronic active enteritis with pyloric gland metaplasia. The neoplastic epithelium showed diffuse and strong nuclear immunoreactivity for p53. The presence of lamina propria deep to the dysplastic epithelium-lined cysts/diverticula supported a non-invasive nature of this lesion. No metastatic disease was noted in 15 nodes identified. All surgical resections were free of neoplasm. The patient was doing well 9 months after surgery without evidence of cancer recurrence or metastasis on abdominal computed tomography. J Med Cases. 2016;7(6):202-207 doi: https://doi.org/10.14740/jmc2415w

The Relationship between p53 Expression and Human Papillomavirus in Premalignant and Malignant Uterine Cervical Lesions

Objective: We aimed to retrospectively examine a series of premalignant and malignant cervical tissues to study a high-risk HPV 16 infection that, among cervical tissue lesions, carries the greatest risk of conversion to cancer, and the presence of p53 protein immunoreactivity, a tumor suppressor gene product. Methods: Paraffin blocks were studied via immunohistochemical (IHC) method to explore the presence of HPV 16 in 59 premalignant and malignant cervical lesions as well as immunoreactivity of the p53 oncoprotein, the most common cellular tumor suppressor gene product in human cancers. Results: In our series, mutant p53 positivity rate was 35.3% for low-grade CIL, 40% for high-grade CIL, and 46.8% for invasive carcinoma cases. Immune staining for high-risk HPV 16 type yielded a positive staining rate of 47% in low-grade CIL, 80% in high-grade CIL, and 50% in invasive carcinoma. Conclusion: The results of our study indicate a progressive increase in p53 oncoprotein reactivity from cervical intraepithelial neoplasia to invasive carcinoma. This suggests the clinical importance of p53 immunoreactivity in dysplastic progression and neoplastic transformation. HPV is the most commonly encountered oncogenic type in cervical lesions, especially in high-grade CIL and invasive carcinomas. Results of the previous reports suggest that HPV-positive carcinomas release wild type p53 and HPV-negative ones release mutant type p53 were not confirmed by our results, which indicated a mutant type p53 reactivity in HPV- 16 positive carcinoma cases.

Detection of human papillomavirus infection in oral squamous cell carcinoma: a cohort study of Japanese patients.

Recent evidence suggests that human papillomavirus (HPV)-related head and neck squamous cell carcinoma (HNSCC) is a separate HNSCC subgroup with distinct epidemiology, histopathological characteristics, therapeutic response to chemotherapy and radiation, and clinical outcome. This study aimed to investigate the role of HPV infection in oral squamous cell carcinoma (OSCC) and the correlation between HPV infection, tumor suppressor protein p16 expression, and clinicopathological features in Japanese patients. In total, 174 OSCC specimens were examined for p16 levels by immunohistochemistry, and p16-positive OSCCs were analyzed for HPV DNA by in situ hybridization (ISH) and HPV genotypes by real-time PCR. The results were evaluated for the association with clinicopathological characteristics of OSCC patients. Twenty-four OSCC samples were found positive for p16 expression; all of them were well-differentiated tumors. P16 immunoreactivity was significantly associated with the invasion depth and tended to correlate with sex, site in the oral cavity, stromal reaction, TNM stage, and survival. HPV DNA was detected in 13 of 24 (54%) p16-positive OSCC by real-time PCR; HPV 16, 18, and other high-risk genotypes were the most prevalent. However, ISH failed to detect HPV DNA in p16-positive OSCCs. P16 immunoreactivity and HPV genotyping by real-time PCR may be useful markers of HPV infection in OSCC. However, although HPV-related OSCC showed good outcomes, HPV infection may have a minor role in oral oncogenesis in Japanese patients.

Regulation of p53 under hypoxic and inflammatory conditions in periodontium.

Different studies suggest that inflammation as well as hypoxia leads to an increase of p53 protein levels. However, the implication of p53 during oral inflammatory processes is still unknown. The aim of this study was therefore to investigate the effect of hypoxia and inflammation on p53 regulation in human periodontium in vitro and in vivo. Under hypoxic and normoxic conditions, human primary periodontal ligament (PDL) fibroblasts (n=9) were stimulated with lipopolysaccharides (LPS) from Porphyromonas gingivalis (P.g.), a periodontal pathogenic bacterium. After different time points, cell viability was tested; p53 gene expression, protein synthesis, and activation were measured using quantitative RT-PCR, immunoblotting, and immunofluorescence. Moreover, healthy and inflamed periodontal tissues were obtained from 12 donors to analyze p53 protein in oral inflammatory diseases by immunohistochemistry. LPS-P.g. and hypoxia initially induced a significant upregulation of p53 mRNA expression and p53 protein levels. Nuclear translocation of p53 after inflammatory stimulation supported these findings. Hypoxia first enhanced p53 levels, but after 24h of incubation, protein levels decreased, which was accompanied by an improvement of PDL cell viability. Immunohistochemistry revealed an elevation of p53 immunoreactivity in accordance to the progression of periodontal inflammation. Our data indicate that p53 plays a pivotal role in PDL cell homeostasis and seems to be upregulated in oral inflammatory diseases. Upregulation of p53 may promote the destruction of periodontal integrity. A possible relationship with carcinogenesis may be discussed.

Expression of pRb and p16INK4 in human thymic epithelial tumors in relation to the presence of human polyomavirus 7.

BackgroundWe have recently reported the presence of the Human polyomavirus 7 (HPyV7) in human thymic epithelial tumors as assessed by diverse molecular techniques. Here we report on the co-expression of p16, retinoblastoma protein (pRb) and phosphorylated retinoblastoma protein (phospho-Rb) in human thymic epithelial tumors in relation to HPyV7.MethodsPRB, phospho-RB and p16 expression was assessed by immuno-histochemistry in 37 thymomas and 2 thymic carcinomas. 17 thymomas (46 %) and 1 thymic carcinoma (50 %) were recently tested positive for HPyV7. In addition, 20 follicular hyperplasias were tested.ResultsExpression of pRb was observed in 35 thymomas (94.6 %), in 16 thymomas (43.2 %) the expression was strong. Phospho-Rb was observed in 31 thymomas (83.8 %). 19 thymomas (51.4 %) showed immunoreactivity for p16 of which 8 thymomas revealed very strong p16 expression. No p16 expression was detected in thymic carcinomas. In addition, no significant correlation between the presence of HPyV7 and pRb-, phospho-Rb- and p16-expression could be established. No correlation between pRb, phospho-Rb, p16 and WHO staging, Masaoka-Koga staging or the presence of MG was found. All 20 follicular hyperplasias showed expression of pRb and less expression of phospho-Rb.ConclusionsAlthough polyomaviruses have been shown to interact with cell cycle proteins no correlation between the presence of HPyV7 and the expression of pRb, phospho-Rb and p16 in human thymic epithelial tumors was observed. In as much HPyV7 contributes to human thymomagenesis remains to be established. Our data indicate pRb, phospho-Rb and p16 expression are rather unlikely to be involved in HPyV7 related thymomagenesis.

P16INK4A expression is frequently increased in periorbital and ocular squamous lesions.

Backgroundp16 expression is a well established biomarker of cervical dysplasia and carcinoma arising from high risk human papilloma virus infection. Increased p16 expression is also seen in squamous neoplasms arising at other sites, including head, neck, and oropharyngeal tract. Squamous lesions are also frequently encountered at ocular surface and peri-orbital skin sites, but the prevalence of increased p16 expression in these lesions has been poorly studied.MethodsWe retrospectively surveyed 13 ocular surface and 16 orbital squamous lesions biopsied at UC San Diego Healthcare System and VA San Diego Healthcare System for p16 expression by immunohistochemistry. These cases included ocular surface lesions with diagnoses of conjunctival intraepithelial neoplasm (CIN) and squamous cell carcinoma in situ. Peri-orbital eyelid biopsies included lesions with diagnoses of SCCis and invasive squamous cell carcinoma. We performed multivariate logistic regression, followed by student’s T-test or Fisher's exact test to determine if there were statistically significant associations between p16 immunoreactivity and patient age, gender, diagnosis, and ethnicity. Statistical significance was defined as p < 0.05.ResultsWe found an unexpectedly large prevalence of strong nuclear and cytoplasmic p16 immunoreactivity in our cases. Almost all of the ocular surface squamous lesions were diffusely positive for p16 expression (12/13). All of the periorbital lesions showed diffuse p16 immunoreactivity (16/16). Altogether, 28/29 lesions tested showed strong and diffuse p16 expression. We found no statistically significant correlation between p16 expression and patient age, gender, ethnicity, or diagnosis. In 6 of the peri-orbital biopsies, we had sufficient tissue to assess high-risk HPV expression by in situ hybridization. Interestingly, all of these cases were negative for HPV, despite strong p16 expression.ConclusionStrong p16 expression was observed in virtually all of the ocular surface and peri-orbital squamous neoplasms in our study. The relationship between p16 expression and HPV infection in ocular surface and peri-orbital sites requires further investigation.

Expression of P40 and P63 in lung cancers using fine needle aspiration cases. Understanding clinical pitfalls and limitations

Fine-needle aspiration (FNA) biopsy of lung lesions is a highly accurate method for diagnosing and staging of lung cancers, particularly in patients with advanced cancer. Although, the majority of FNA cases of non-small cell lung carcinoma (NSCLC) can be subclassified by hematoxylin and eosin (H&E) sections, immunohistochemical (IHC) markers are usually necessary for difficult cases. Our previous study has shown that both P40 and P63 demonstrate differential sensitivity and specificity in the subclassification of squamous cell carcinoma (SqCC) using tumor tissue microarrays (TMA). In the present study, we further evaluated the utility of P40 and P63 and the potential pitfalls and limitations associated with the usefulness of these stains in FNA cases. By a computer search of pathology archives, 144 FNA biopsies with diagnoses of lung cancers and P40/P63 stains were identified, including 50 adenocarcinomas (ADCs), 56 SqCCs, 8 small cell lung carcinomas (SCLCs), and 12 cases of poorly differentiated carcinoma (PD CA). Ten benign FNA lung lesions and 8 other malignant neoplasms were also included as controls. Nuclear staining patterns of P40 and P63 were scored semi-quantitatively as 0 (negative), 1 (<10%, weak and focal), or 2 (>10%, strong and diffuse). In lung SqCCs, P40 and P63 were positive in 77.3% and 89.5% cases, respectively. In ADCs, P40 was weakly and focally positive in 6.1% cases, and P63 was variably positive in 62.8% cases. In SCLCs, P40 and P63 were focally positive in 12.5% and 50% cases. In PD CAs, no P40 or P63 immunoreactivity was detected. In the group of other neoplasms (n=8) both P40 and P63 were positive in the case of metastatic non-seminomatous germ cell tumor (NSGCT) (n=1), and P63 was positive in the case of metastatic Merkel cell carcinoma (n=1). The sensitivity and specificity of P40 and P63 were 76.9%/93.3%, and 90.2%/50.7% in the lung SqCC. P63 has a better sensitivity, and P40 has a better specificity for SqCC. A positive staining pattern with both markers was also found in certain non-SqCC cases. Recognizing limitations of these markers are particularly important in FNA cases.

Abstract 3460: Fetal alcohol exposure increases susceptibility to carcinogenesis in the pituitary

Abstract The idea that exposure to adverse environmental conditions and lifestyle choices during pregnancy can result in fetal programming that underlies disease susceptibility in adulthood is now widely accepted. Fetal alcohol exposed offspring display many behavioral and physiological abnormalities including neuroendocrine-immune functions, which often carry over into their adult life. Since neuroendocrine-immune system is critically involved in the regulation of tumor surveillance, we determined whether fetal alcohol exposure increases the susceptibility to estrogen-induced pituitary prolactin-secreting tumors (prolactinomas), commonly occurring pituitary tumors in humans. Pregnant Fischer 344 rats were fed between gestational days 7 and 21 with a liquid diet containing alcohol (AF), pair-fed with isocaloric liquid diet (PF), or fed ad libitum with rat chow (AD). At 90 days of age, female offspring rats were ovariectomized and received a subcutaneous estradiol implant. These rats were sacrificed between 4 and 5 months after the estradiol implants. At the time of sacrifice, pituitaries of these animals were inspected for tumor and whole body were inspected for any tumor metastasis. Estradiol treatment increased pituitary weight about 5-folds in AD and PF treated groups and increased 20-30-folds in the AF treated group. Most tumors in the AF group were hemorrhagic. About 30% AF rats had some tumors in the non-pituitary sites. AD and PF rats did not show any non-pituitary site tumor. When AF rat pituitaries were grown in cultures, cells rapidly grew and formed colonies. Colony formation and rapid growth rate were not observed in cells of the pituitary from PF and AD rats. Histopathological evaluation revealed that tumors in AF group were more densely packed cells as compare to the PF and AD groups who showed uniform cells with abundant cytoplasm. Pituitary tumor from alcohol exposed animals showed strong nuclear p53 and monoclonal Ki67 immunoreactivity in almost all tumor cells. Significantly higher mRNA levels of hemorrhage-associated genes (VEGF and MMP-9) were also observed in pituitary tumor tissues from AF group as compare with PF and AD groups. Tumor cells showed positivity to prolactin staining but showed negativity to other pituitary hormones staining. Histological evaluation of the pituitary tissues revealed that tumors were prolactin producing in all groups, and tumors in AF pituitaries were highly proliferative. These data provide evidence for aggressive, possible neoplastic, prolactinoma development in the pituitary after estrogen treatment in fetal alcohol exposed female rats. (This work is supported by a National Institute of Health grant R01 AA11591). Citation Format: Shaima Jabbar, George Maglakelidze, Dipak K. Sarkar. Fetal alcohol exposure increases susceptibility to carcinogenesis in the pituitary. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3460. doi:10.1158/1538-7445.AM2015-3460

Abstract 4000: The increase of oncogenic miRNA expression in tongue carcinogenesis of a mouse model

Abstract Oral carcinoma is one of the leading causes of cancer death in the world. To develop biomarkers for early detection may bestow substantial benefits in diagnosis and the prevention of tumor progression. An animal model simulating the human pathogenesis will also be useful for developing therapeutic strategies. miRNAs are short, non-coding RNAs, which play important roles in neoplastic process. The detection of biomarkers in body fluid may enable non-invasive diagnostic approaches. This study used mouse chemical carcinogenesis model to investigate the disruption of miRNA expression in oral tongue tissues, and to explore the miRNA level in saliva and blood during pathogenesis. Mice were fed with 4-Nitroquinoline 1-Oxide (4NQO) in drinking water for 12 weeks and were kept for extended periods for tumor induction. The histological examination showed the consecutive increase in the severity of epithelial pathogenesis from hyperkeratosis, hyperplasia to epithelial dysplasia and squamous tumors following the increase of 4NQO exposure. The increase of miR-21 and miR-31 staining, as well as p53, pAKT and Ki-67 immunoreactivity parallels the severity of epithelial pathogenesis in tongue tissues. A progressive increase of multiple oncogenic miRNAs in both saliva and blood samples was also noted. The increase of miR-21, miR-31 and miR-372 in the saliva and the increase of miR-146a and miR-184 in the blood sample were particularly eminent during the field cancerization process of tongue. In addition, the dysregulation of these oncogenic miRNAs and other molecular events in this model are similar to the disruption being identified in human counterparts. As these miRNAs are functionally oncogenic, their increase in the saliva and blood of mice may be valuable biomarkers to develop diagnosis and prevention strategies against oral carcinomas. Keywords: chemical, carcinoma, miRNA, mouse, mouth Note: This abstract was not presented at the meeting. Citation Format: YUYU KAO, Shu-Chun Lin, Kuo-Wei Chang. The increase of oncogenic miRNA expression in tongue carcinogenesis of a mouse model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4000. doi:10.1158/1538-7445.AM2015-4000

Clinicopathological and immunohistological features of polypoid endometriosis.

To compare clinicopathological and immunohistochemical features of polypoid endometriosis (PE) and non-polypoid endometriosis (NPE). Fifteen cases of PE and 20 cases of NPE were assessed. All cases were stained immunohistochemically for CD10 and p16 and the proportion of p16-positive stromal and epithelial cells was estimated. On review, 10 PE cases resembled NPE histologically but occurred at mucosal or serosal surfaces, or within cyst cavities, that permitted polyp formation. These cases had a similar age distribution and immunohistochemical profile to NPE. The remaining five PE cases showed histological similarity to eutopic endometrial polyps; these occurred in older patients, and showed significantly greater stromal and epithelial p16 immunoreactivity. There are two main subgroups of PE. The majority of cases in this series showed similar histological features to NPE, but involved anatomical sites that facilitated exophytic or polypoid growth. The remaining PE cases resembled eutopic endometrial polyps histologically and immunophenotypically and they occurred in older patients. These findings suggest that such lesions are 'true' polyps sharing a pathogenetic relationship with similar lesions arising in the endometrium.

Endoscopic ultrasound-guided fine-needle aspiration cytology in pancreaticobiliary carcinomas: diagnostic efficacy of cell-block immunocytochemistry

Endoscopic ultrasound-guided fine-needle aspiration cytology was demonstrated to be a useful tool for the diagnosis and staging of pancreaticobiliary neoplastic lesions. Nonetheless, the diagnostic value of this procedure may be limited by low cellularity of the specimen, contamination of intestinal cells and unfeasibility of ancillary immunocytochemical procedures. The present study was to evaluate its usefulness in the diagnosis of neoplastic lesions. A series of 46 pancreaticobiliary carcinomas with available cell block preparations was submitted to immunocytochemistry against cytokeratins, carcinoembryonic antigen, E-cadherin, CD10 and p53. The sensitivity, specificity, positive and negative predictive values of the cytological smear in the discrimination of malignant lesions were calculated and compared with those of cell block preparation with the immunocytochemical stains against p53 and CD10. According to our findings, the use of cell block preparations together with immunostains against p53 and CD10 allowed to discriminate malignant versus benign specimens with higher sensitivity than the only cytological examination. In detail, CD10 immunostaining was of significant help for the discrimination between cytological contaminants, such as benign gastrointestinal cells, and the neoplastic elements of pancreaticobiliary well differentiated adenocarcinomas. Also, intense nuclear immunoreactivity for p53 was encountered in about 2/3 of the cases and identified pancreatic malignancy with high sensitivity. We suggest that immunocytochemistry against both CD10 and p53 could be applied case by case, mainly to differentiate gastrointestinal and pancreatic benign cellular contaminants showing hyperplasia or reactive changes from differentiated pancreaticobiliary adenocarcinomas.

Comparative analysis of p53 and p21 proteins in normal cervix and HPV associated precancerous and cancerous lesions of cervix

IntroductionHPV-E6 oncoprotein of High-Risk HPV plays critical role in the degradation of p53 protein for acquisition of malignant phenotype in cervix cells. The present study, was sought to analyze the expression of HPV-E6, p53 and p21 in cervical cancer to explore possible impact of HPV-E6 in the modulation of these proteins. MethodsThe expression of HPV16/18-E6, p53 and p21 proteins in cervix tissues [normal (n = 100), cervical intraepithelial neoplasia (CIN; n = 67), squamous cell carcinoma (SCCs; n = 153)] was analyzed by immunohistochemistry and revalidated by Western Blotting. The p53 protein functionality was determined by using phosphospecific p-p53-ser46 antibody. ResultsThe negative expression of HPV-E6 and p53 but, mild immunoreactivity of p21 was noticed in normal stratified squamous epithelium. In CIN and SCCs, the HPV-E6 was found to be predominantly over-expressed in 80.0% and 84.9% of cases respectively. Significant nuclear accumulation of p53 was observed in 77.6% of precancerous and 83.0% of cancerous tissues whereas, 74.6% of CIN and 73.8% of SCCs represented loss of nuclear p21 expression as compared to normal (p = 0.0001, p = 0.0001). Pearson's correlation test revealed significant inverse association between p53 and p21 proteins both in CIN (p = 0.0001) and SCCs (p = 0.0001). Interestingly, significant positive association of HPV-E6 with p53 and negative association with p21 were also detected in CIN (p = 0.0001) and SCCs (p = 0.0001). No positive expression of p-p53-ser46 was detected in any of the cases. DiscussionThe present study provides the evidences of inhibition of p53 and p21 transactivity by HPV-E6 resulting alteration of cell growth inhibitory signals in cervical cancer.

Carcinogenic HPV infection in the cervical squamo-columnar junction.

Recent studies have suggested the involvement of a unique population of cells at the cervical squamo-columnar junction (SCJ) in the pathogenesis of early (squamous intraepithelial lesion or SIL) and advanced (squamous cell and adeno-carcinomas) cervical neoplasia. However, there is little evidence to date showing that SCJ cells harbour carcinogenic HPV or are instrumental in the initial phases of neoplasia. This study was designed to (1) determine if normal-appearing SCJ cells contained evidence of carcinogenic HPV infection and (2) trace their transition to early SIL. Sections of cervix from high-risk reproductive age women were selected and SCJ cells were analysed by using several techniques which increasingly implicated HPV infection: HPV DNA (genotyping and in situ hybridization)/RNA (PCR), immunostaining for HPV16 E2 (an early marker of HPV infection), p16(ink4), Ki67, and HPV L1 protein. In 22 cases with a history of SIL and no evidence of preneoplastic lesion in the excision specimen, HPV DNA was isolated from eight of ten with visible SCJ cells, six of which were HPV16/18 DNA-positive. In five of these latter cases, the SCJ cells were positive for p16(ink4) and/or HPV E2. Transcriptionally active HPV infection (E6/E7 mRNAs) was also detected in microdissected SCJ cells. Early squamous atypia associated with the SCJ cells demonstrated in addition diffuse p16(ink4) immunoreactivity, elevated proliferative index, and rare L1 antigen positivity. We present for the first time direct evidence that normal-appearing SCJ cells can be infected by carcinogenic HPV. They initially express HPV E2 and their progression to SIL is heralded by an expanding metaplastic progeny with increased proliferation and p16(ink4) expression. Whether certain SCJs are more vulnerable than others to carcinogenic HPV genotypes and what variables determine transition to high-grade SIL remain unresolved, but the common event appears to be a vulnerable cell at the SCJ.

TP53 mutations, tetraploidy and homologous recombination repair defects in early stage high-grade serous ovarian cancer.

To determine early somatic changes in high-grade serous ovarian cancer (HGSOC), we performed whole genome sequencing on a rare collection of 16 low stage HGSOCs. The majority showed extensive structural alterations (one had an ultramutated profile), exhibited high levels of p53 immunoreactivity, and harboured a TP53 mutation, deletion or inactivation. BRCA1 and BRCA2 mutations were observed in two tumors, with nine showing evidence of a homologous recombination (HR) defect. Combined Analysis with The Cancer Genome Atlas (TCGA) indicated that low and late stage HGSOCs have similar mutation and copy number profiles. We also found evidence that deleterious TP53 mutations are the earliest events, followed by deletions or loss of heterozygosity (LOH) of chromosomes carrying TP53, BRCA1 or BRCA2. Inactivation of HR appears to be an early event, as 62.5% of tumours showed a LOH pattern suggestive of HR defects. Three tumours with the highest ploidy had little genome-wide LOH, yet one of these had a homozygous somatic frame-shift BRCA2 mutation, suggesting that some carcinomas begin as tetraploid then descend into diploidy accompanied by genome-wide LOH. Lastly, we found evidence that structural variants (SV) cluster in HGSOC, but are absent in one ultramutated tumor, providing insights into the pathogenesis of low stage HGSOC.

Involvement of caspases and their upstream regulators in myocardial apoptosis in a rat model of selenium deficiency-induced dilated cardiomyopathy

Keshan disease is an endemic dilated cardiomyopathy (DCM) which is closely related with selenium-deficient diet in China. In the previous study, we reported that the low selenium status plays a pivotal role in the myocardial apoptosis in the DCM rats, however, the underlying mechanism remains unclear. The present study aimed to determine whether the intrinsic, extrinsic pathways and the upstream regulators were involved in the myocardial apoptosis of selenium deficiency-induced DCM rats. Therefore, the rat model of endemic DCM was induced by a selenium-deficient diet for 12 weeks. Accompanied with significant dilation and impaired systolic function of left ventricle, an enhanced myocardial apoptosis was detected by TUNEL assay. Western blot analysis showed remarkably increased protein levels of cleaved caspase-3, caspase-8, caspase-9, and cytosolic cytochrome c released from the mitochondria. In addition, the immunoreactivities of p53 and Bax were significantly up-regulated, while the anti-apoptotic Bcl-2 family members Bcl-2 and Bcl-XL were down-regulated. Furthermore, appropriate selenium supplement for another 4 weeks could partially reverse all the above changes. In conclusion, the intrinsic, extrinsic pathways and the upstream regulators such as p53, Bax, Bcl-2, and Bcl-XL were all involved in selenium deficiency-induced myocardial apoptosis.

Correlation of p53 immunoexpression with DNA ploidy and apoptotic index in subsets of prostate cancer: A marker reiterated in progression and recurrence of prostate cancer.

Background:Prediction of biological behavior in patients of prostate cancer (CaP) is a major challenge as current parameters only partially meet the need for prognostication. p53 as a prognostic indicator has been studied in several human cancers, including breast, lung, and colorectal carcinoma. However, its significance as a predictive biomarker for CaP is less well-studied.Materials and Methods:This study included 125 cases of CaP, 27 cases of prostatic intraepithelial neoplasia and 25 cases of benign prostatic hyperplasia. Immunohistochemical assessment for p53 nuclear protein was performed. Assessment for apoptotic index and DNA ploidy status by flow cytometry were also done.Results:p53 immunoreactivity was low in organ confined CaP cases having Gleason score ≤3 (P < 0.003). More hormone resistant cases 37 (83%) were aneuploid when compared with hormone sensitive cases 26 (33%) (P < 0.005). 93% of p53 positive cases and none of the p53 negative patient were aneuploid suggesting a significant relation between p53 immunoreactivity and aneuploidy. p53 positivity and DNA aneuploidy, independently, were also predictors of progression and relapse.Conclusion:DNA ploidy and p53 positivity go hand in hand and together yield additional prognostic information in CaP. p53 positivity is possibly a late event in carcinogenesis in CaP and a marker of change in biological behavior of CaP.

MP36-06 P63 EXPRESSION IN BLADDER CANCER VERSUS CHRONIC BILHARZIAL BLADDER

You have accessJournal of UrologyBladder Cancer: Basic Research I1 Apr 2015MP36-06 P63 EXPRESSION IN BLADDER CANCER VERSUS CHRONIC BILHARZIAL BLADDER Khaled Mursi, Ayman Agag, Olfat Hammam, Essam Riad, and Mahmoud Daw Khaled MursiKhaled Mursi More articles by this author , Ayman AgagAyman Agag More articles by this author , Olfat HammamOlfat Hammam More articles by this author , Essam RiadEssam Riad More articles by this author , and Mahmoud DawMahmoud Daw More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.734AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To investigate the immunohistochemical expression of p63 in bladder cancer and the variation in this expression in relation to the histological type, grade, and stage of the tumor, and whether bilharziasis (endemic in Egypt) has an effect on its expression or not. METHODS A prospective study conducted on 70 patients divided into 3 groups, Group A: ten patients with normal healthy urothelium, Group B: 20 patients with chronic cystitis (bilharzial and non-bilharzial), and Group C: 40 patients with bladder cancer. Biopsies were taken from all patients and were examined for the expression of p63 by immunohistochemical techniques. RESULTS The mean age (±SD) for the control, cystitis, and the malignant group w as 45.2±9.5, 50.5±11.7, and 60.52±9.9 years respectively. There was a statistically significant decrease in the p63 expression and immunoreactivity in the malignancy group (p<0.05). Also, a significant decrease in the expression was found with the advancement of the tumor stage and grade (p<0.01). In Squamous cell tumors, there was a statistically significant decrease in the p63 immunoreactivity compared to transitional cell tumors (p<0.01). Regarding the impact of bilharziasis, there w as a tendency for the statistically significant decrease in the immunoreactivity in bilharzial cystitis (p<0.01), but in the malignant group, bilharziasis did not seem to affect the pattern of expression. CONCLUSIONS p63 may be a helpful biomarker and adjunct in predicting the tumor biological behavior and progression, further studies are recommended to elucidate exactly its role as a prognostic indicator and its utility as a tumor marker. Nuclear p63 immmunoreactivity Negative (0-10%), n (%) Mild (11-30%), n (%) Moderate (31-60%), n (%) Strong (61-100%), n (%) Total positive, n (%) Control, n=10 0 0 0 10 (100%) 10 (100%) Chronic cystitis, n=20 Non-bilharzial, n=10 0 0 0 10 (100%) 10 (100%) Bilharzial, n=10 0 0 3 (30%)# 7 (70%)# 10 (100%) Total, n=20 0 0 3 (15%)∗ 17 (85%)∗ 20 (100%) Malignant lesions, n=40 TCC, n=35 4 (16%) 5 (20%)ˆ 4 (16%)ˆ 12 (48%)ˆ 21 (84%) Superficial TCC, n=12 1 (8.3%) 1 (8.3%) 3 (25%) 7 (58.4%) 11 (91.7%) Invasive TCC, n=13 3 (23%)ˆ ˆ 4 (30.8%)ˆ ˆ 1 (7.7%)ˆ ˆ 5 (38.5%)ˆ ˆ 10 (76.9%)ˆ ˆ SqCC, n=15 3 (20%) 7 (46.7%) 1 (6.7%) 4 (26.6%) 12 (80%) Total, n=40 7 (17.5%)∗ 12 (30%)∗∗ 5(12.5%)∗ 16 (40%)∗∗ 33 (82.5%) Stage, n=40 Ta, n=5 0 0 1 (20%) 4 (80%) 5 (100%) T1, n=7 1 (14.3%) 1 (14.3%) 2 (28.6%) 3 (42.8%)† 6 (85.5%)† T2 - T3, n=28 6 (21.4%)‡ 11 (39.3%)‡ 2 (7.2%)†‡ 9 (32.1%)†‡ 22 (78.5%)† Grade, n=40 Grade 1, n=8 0 (0%) 1 (12.5%) 1 (12.5%) 6 (75%) 8 (100%) Grade 2, n=23 3 (13%)+ 9 (39%)+ 3 (13%) 8 (65%) 20 (87%) Grade 3, n=9 4 (44.5%)+± 2 (22.2%)+± 1 (11.1%) 2 (22.2%)+± 5 (55.5%)+± Malignant, n=40 Non-bilharzial, n=10 3 (30%) 3 (30%) 2 (20%) 2 (20%) 7 (70%) Bilharzial, n=30 4 (13.3%)& 9 (30%) 3 (10%)& 14 (46.7%)& 26 (86.6%)& # p< 0.01 compared to chronic non-bilharzial cystitis ∗ p< 0.05 compared to control cases ∗∗ p <0.01 compared to control cases ˆ p< 0.01 compared to SqCC cases ˆˆ p< 0.01 compared to superficial TCC cases † p< 0.01 compared to Ta ‡ p< 0.01 compared to T1 + p< 0.01 compared to Grade 1 ± p< 0.01 compared to Grade 2 & p< 0.01 compared to non-bilharzial malignant lesion. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e430-e431 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Khaled Mursi More articles by this author Ayman Agag More articles by this author Olfat Hammam More articles by this author Essam Riad More articles by this author Mahmoud Daw More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Murine double minute 2 predicts response of advanced esophageal squamous cell carcinoma to definitive chemoradiotherapy.

BackgroundDefinitive chemoradiotherapy (dCRT) has recently become one of the most effective therapies for the treatment of esophageal squamous cell carcinoma (ESCC). However, it is also true this treatment has not been effective in all patients. Therefore, it is very important to evaluate the surrogate marker of dCRT in order to improve clinical outcomes of patients with ESCC. On the other hand, our previous study had suggested that murine double minute 2 (MDM2) and p16 were associated with chemoradioresistance in ESCC.MethodsWe selected pretreatment biopsy specimens of ESCC patients from our prospective clinical study on dCRT. Seventy-nine cases histologically diagnosed as ESCC were used. We immunohistochemically investigated these specimens using antibodies against MDM2, p53, p16, and Ki-67.ResultsThe patients included 68 males and 11 females with a mean age of 63.3 years. The number of patients in each clinical stage was as follows: 22 in c-Stage I; 17 in c-Stage II; and 40 in c-Stage III. cT, cN, and cStage were significantly more advanced in the Failure group (including patients with persistent and recurrent disease after dCRT) than in the complete response (CR) group (patients with persistent CR after dCRT). The clinical stage inversely correlated with the CR rate and the rescue rate after failure. The overall survival rate was significantly worse in the patients with advanced cT, cN, and cStage levels, and in the Failure group. MDM2 positivity was significantly higher in the Failure group than in the CR group in cStageIII (P = 0.014). The number of patients with an absence of p16 immunoreactivity was significantly higher in the Failure group than in the CR group in cStageIII (P = 0.010) but not in cStageI or cStageII. Moreover, the overall survival with a Ki-67 ≥ 33.7% was significantly better than that with <33.7% for patients in cStageIII (P = 0.024).ConclusionsThe results of this study suggested that MDM2 and p16 are predictive markers for chemoradioresistance in cStageIII ESCC and Ki-67 is a prognostic marker following dCRT in cStageIII ESCC. These issues could contribute to the formulation of treatment strategy for patients with advanced ESCC.

P63 Expression in the Gerbil Hippocampus Following Transient Ischemia and Effect of Ischemic Preconditioning on p63 Expression in the Ischemic Hippocampus.

p63 is a transcription factor of p53 gene family, which are involved in development, differentiation and cell response to stress; however, its roles in ischemic preconditioning (IPC) in the brain are not clear. In the present study, we investigated the effect of IPC on p63 immunoreactivity caused by 5 min of transient cerebral ischemia in gerbils. IPC was induced by subjecting the gerbils to 2 min of transie ischemia 1 day prior to 5 min of transient ischemia. The animals were randomly assigned to four groups (sham-operated-group, ischemia-operated-group, IPC plus (+)-sham-operated-group and IPC + ischemia-operated-group). The number of viable neurons in the stratum pyramidale of the hippocampal CA1 region (CA1) was significantly increased by IPC + ischemia-operated-group compared with that in the ischemia-operated-group 5 days after ischemic insult. We found that strong p63 immunoreactivity was detected in the CA1 pyramidal neurons in the sham-operated-group, and the immunoreactivity was decreased with time after ischemia-reperfusion. In addition, strong p63 immunoreactivity was newly expressed in microglial cells of the CA1 region from 2 days after ischemia-reperfusion. In all the IPC + sham-operated-groups, p63 immunoreactivity in the CA1 pyramidal neurons was similar to that in the sham-operated-group, and the immunoreactivity was well maintained in the IPC + ischemia-operated-groups after cerebral ischemia. In brief, our present findings show that IPC dramatically protected the reduction of p63 immunoreactivity in the pyramidal neurons of the CA1 region after ischemia-reperfusion, and this result suggests that the expression of p63 may be necessary for neurons to survive after transient cerebral ischemia.

P53, KI-67, CD117 expression in gastrointestinal and pancreatic neuroendocrine tumours and evaluation of their correlation with clinicopathological and prognostic parameters.

Gastrointestinal and pancreatic neuroendocrine tumors (GEPNETs) originate from the cells of the endocrine system. Their molecular genetic mechanism of development and progression is complex and remains largely unknown. The purpose of this study was to review the gastrointestinal and pancreatic neuroendocrine tumors and to evaluate p53, Ki-67 and CD 117 expressions with their clinicopathological correlations. Twenty-one patients were reviewed and classified as having well-differentiated neuroendocrine neoplasm (WDET, Grade I), well-differentiated neuroendocrine carcinoma (WDEC, Grade II) and poorly differentiated neuroendocrine carcinoma (PDEC, Grade III). We performed immunohistochemical tests to characterize the expession of the immunoreactivity for synaptophysin, chromogranin, p53, Ki67 and CD 117. Median age of 21 patients was 43 years. Thirteen (61.9%) patients were male and eight (38.1%) patients were female. Tumors were located in the stomach (38.1%), appendix (38.1%), duodenum (4.8%), ileum (4.8%), colon (9.5%), and pancreas (4.8%). There was a statistically significant difference between well-differentiated endocrine neoplasm (Grade I), and well-differentiated endocrine carcinoma (WDEC, Grade II) and PDEC for Ki-67 >20% (p<0.001) (Pearson chi-square test). There was a statistically significant difference between WDET (Grade I), WDEC (Grade II) and PDEC (Grade III) for p53 positivity (p<0.05) (Pearson chi-square test).