Nassim Saremi

Significance of PI3K/AKT signaling pathway in metastasis of esophageal squamous cell carcinoma and its potential as a target for anti-metastasis therapy

Metastasis is the most lethal hallmark of esophageal squamous cell carcinoma (ESCC). The aim of the study is to identify key signaling pathways that control metastasis in ESCC. Highly invasive ESCC sublines (designated I3 cells) were established through three rounds of selection of cancer cells invading through matrigel-coated chambers. Gene expression profile of one of the I3 sublines was compared with that of its parental cell line using cDNA microarray analysis. Gene ontology and pathway analyses of the differentially expressed genes (both upregulated and downregulated) indicated that genes associated with cellular movement and the AKT pathway were associated with increased cancer cell invasiveness. Western blot analysis confirmed increased phosphorylated AKT (p-AKT), N-cadherin and decreased E-cadherin expression in the I3 cells. Immunohistochemistry was used to evaluate the clinical significance of p-AKT expression in ESCC, and the results showed higher p-AKT nuclear expression in lymph node metastases when compared with primary carcinoma. Inactivation of the PI3K/AKT pathway with specific inhibitors, or with PTEN overexpression, resulted in reversed cadherin switching and inhibited cancer cell motility. Inhibition of the pathway by treatment with wortmannin markedly suppressed experimental metastasis in nude mice. Our data demonstrated the importance of the PI3K/AKT signaling pathway in ESCC metastasis and support PI3K/AKT as a valid therapeutic target in treatment of metastatic ESCC.

“Googling” for cancer: An infodemiological assessment of online search interests in Australia, Canada, New Zealand, the United Kingdom, and the United States

Background The infodemiological analysis of queries from search engines to shed light on the status of various noncommunicable diseases has gained increasing popularity in recent years. Objective The aim of the study was to determine the international perspective on the distribution of information seeking in Google regarding “cancer” in major English-speaking countries. Methods We used Google Trends service to assess people’s interest in searching about “Cancer” classified as “Disease,” from January 2004 to December 2015 in Australia, Canada, New Zealand, the United Kingdom, and the United States. Then, we evaluated top cities and their relative search volumes (SVs) and country-specific “Top searches” and “Rising searches.” We also evaluated the cross-country correlations of SVs for cancer, as well as rank correlations of SVs from 2010 to 2014 with the incidence of cancer in 2012 in the abovementioned countries. Results From 2004 to 2015, the United States (relative SV [from 100]: 63), Canada (62), and Australia (61) were the top countries searching for cancer in Google, followed by New Zealand (54) and the United Kingdom (48). There was a consistent seasonality pattern in searching for cancer in the United States, Canada, Australia, and New Zealand. Baltimore (United States), St John’s (Canada), Sydney (Australia), Otaika (New Zealand), and Saint Albans (United Kingdom) had the highest search interest in their corresponding countries. “Breast cancer” was the cancer entity that consistently appeared high in the list of top searches in all 5 countries. The “Rising searches” were “pancreatic cancer” in Canada and “ovarian cancer” in New Zealand. Cross-correlation of SVs was strong between the United States, Canada, and Australia (>.70, P<.01). Conclusions Cancer maintained its popularity as a search term for people in the United States, Canada, and Australia, comparably higher than New Zealand and the United Kingdom. The increased interest in searching for keywords related to cancer shows the possible effectiveness of awareness campaigns in increasing societal demand for health information on the Web, to be met in community-wide communication or awareness interventions.

Enhancing pathology learning experience of medical students using multiple advanced learning and teaching strategies.

Aim: Currently most medical schools use an integrated multidisciplinary approach in their curricula; therefore creating a huge challenge to engage medical students by delivering traditional pathology modules. In this study, we aimed to implement multiple advanced strategies to improve medical students’ pathology learning experience at Griffith University. Methods: Students enrolled in the second year of Griffith medical programme between 2011 and 2012 were invited to complete questionnaires rating the value and impact of resources delivered on their learning experience. In total, 272/ 290 students responded. The strategies adopted include virtual microscopy, web-based digitalised interactive modules, clinical scenario-integrated lectures, practical histology sessions and gross specimen demonstration. Quality and usefulness of the delivery of these modules were assessed using a 5 scale questionnaire. Results: In both years, overall score was high (mean score >4.5/ 5) for the histology lectures, clinical integrations and virtual microscopy sessions. The traditional delivery of practical and lecture sessions received lower scores. Qualitative comments suggested that the advanced methods were extremely useful for students’ learning experience of pathology. Discussion: A multidisciplinary approach by clinico-pathological integration and the use of virtual microscopy has the potential to better engage students to pathology learning in the modern medical curriculum.

Application of Tissue Microarray in Esophageal Adenocarcinoma

Tissue microarray technology could allow immunohistochemical staining or in situ hybridization on hundreds of different tissue samples simultaneously. It allows faster analysis and considerably reducing costs incurred in staining. The technique also provides a high-throughput analysis of multiple tissues for the different types of research. In the literature, many researches of esophageal adenocarcinoma use tissue microarray to enhance the output. In this chapter, we have a brief overview of tissue microarray technologies, the advantages and disadvantages of tissue microarray, and related troubleshootings.

Bilateral orbital paraganglioma: A report of a unique case and updates on its clinicopathological features

S S161 Compared to the control group, the CDK6 protein level was decreased in OCI-LY1/miR-320d group (p<0.05). OCI-LY1/ miR-320d group and OCI-LY1/CDK6 shRNA group cells’ proliferation ability were markedly decreased compared to the control group (p<0.05). Relative luciferase activity value of experimental group is lower than that of the control group (p<0.05). Conclusions: Up-regulation of miR-320d expression can inhibit the ability of malignant proliferation of DLBCL cell. The mechanism is concerned with that miR-320d could inhibit the expression of target gene CDK6 at the post-transcriptional level. 131. IMPROVED IMMUNOHISTOCHEMICAL METHOD FOR DETECTING ANTI-BCL-2 AND C-MYC ANTIBODIES IN MOUSE XENOGRAFT MODELS Xiaoling Xia, Jean Boyer, Liping Zhang, Burton Holmes, Rajalakshmy Ramalingam, Penny Towne Ventana Medical Systems, Roche, USA Background: Antigen detection is known to be affected by preanalytical conditions such as fixative types, fixation time and delay to fixation. We used human breast carcinoma cell lines (BT-474 and ZR-75-1) generated xenograft tumors as model systems to analyze the impact of different pre-analytical conditions on BCL-2 and c-MYC staining. Antigen detection on mouse tissue is complicated by high level background staining due to the binding of secondary anti-mouse antibody to endogenous mouse tissue Igs and other components. We developed methods to use modified OptiView DAB detection system and unique linker methodology to almost completely eliminate non-specific staining and achieve satisfactory staining results. Methods: Human breast carcinoma cell lines (BT-474 and ZR75-1) xenograft tumors were fixed across a range of times in 6 different commonly used fixatives as well as delay to fixation with a range from immediate fixation to a 24 hour delay to fixation in 10% NBF. Two new IHC staining methods were developed to stain either anti-BCL-2 (124), a mouse monoclonal antibody or anti-cMYC (Y69), a rabbit monoclonal antibody, to compare with standard OptiView DAB detection system on a Ventana automatic staining instrument. An Abcam monoclonal rabbit anti-mouse IgG was employed as linker followed by a modified OptiView DAB system to detect BCl-2 expression. We also used a modified OptiView DAB detection system to measure c-MYC expression. Results: Antigen detection with indirect immunohistochemical methods is hampered by high background staining if the primary antibody is from the same species as the examined tissue. This high background was eliminated by using an Abcam monoclonal rabbit anti-mouse IgG as a linker followed by a modified OptiView DAB detection system. New methods ensured us to achieve a more precise understanding of protein expression level. The xenografts tissues fixed with zinc formalin and Z-5 showed equivalent staining to 10% NBF after fixation for at least 6 hours. However, AFA, 95% EtOH, Prefer fixative did not perform equivalently over the range of fixation times when compared to 10% NBF, regardless of fixation time. This study also demonstrated degradation of BCL-2 and c-MYC antigenicity in response to delay in fixation if tissue was left unfixed at room temperature for more than two hours. Conclusion:Our study recommended that the best fixatives were 10% NBF, Zinc fixative and Z-5 between 6 hours and 72 hours for both anti-BCL-2 and anti-c-MYC antibodies. In addition, the tissues should be fixed within 2 hours after tissue collection. New IHC methods yielded minimum reduction of BCL-2 and c-MYC specific signaling. Using this method, the tissue sections were remarkably free of the background staining that is typically seen in mouse tissues with mouse antibodies. Hence, this procedure provides an improved, high sensitive method for detecting protein expression level loss due to inappropriate pre-analytical sample treatment. 132. RARE HISTOLOGICAL PATTERNS OF ANGIOFIBROMA OF SOFT TISSUE Yuichi Yamada, Hidetaka Yamamoto, Kenichi Kohashi, Yoshinao Oda Department of Anatomic Pathology, Pathological Sciences, Graduate School of Medical Sciences, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka, Japan Background:Angiofibroma of soft tissue (AFST) is a rare benign soft tissue neoplasm, which is characterized by fibroblastic cytomorphology, prominent vascular pattern and novel fusion genes NCOA2-AHRR/AHRR-NCOA2 or GTF2I-NCOA2. AFSTs present a wide spectrum of morphology, making a challenge to diagnosis, thus we came to the cogitation to report the special histological features confirmed by genetic analysis. Aims: The study aims to reveal the histological variety of AFST confirmed by genetic data. Methods: We reviewed all the 274 cases diagnosed as solitary fibrous tumor/hemangiopericytoma (232 cases), undeterminated tumors of fibroblastic differentiation (36 cases) and recently diagnosed as AFST (6 cases), and picked up the 12 cases histologically compatible with AFST. All the 12 cases were genetically analyzed by RT-PCR method, and immunohistochemical stains were performed for available cases. Results: Highlighted histological findings were as follows; amianthoid fibers, ossification, lymphoid follicles, lymphoid cuff, hemosiderin deposition, aggregate of foamy histiocytes, cystic change, necrosis, and hemorrhage. Immunohistochemically, the tumor cells were positive for EMA (4/10 cases), desmin (4/10 cases), CD163 (6/6 cases), CD68 (3/6 cases), estrogen receptor (7/7 cases), progesterone receptor (1/6 case), D2-40 (4/7 cases) and STAT6 (1/6 case, weak nuclear stain), but negative for CD34, alpha-smooth muscle actin, musclespecific actin, S-100 protein, pan-cytokeratin, beta-catenin, MDM2 and CDK4. AHRR-NCOA2 fusion gene was detected in 8 cases. Conclusions: We revealed the unreported histological variation and immunohistochemical findings of AFST and confirmed them by genetic methods. It was suggested that AFST should be considered in the diagnosis of fibrous or fibrohistiocytic tumors with unspecified features.