Alfred K-Y Lam

“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.

Role of miR-193a in Cancer: Complexity and Factors Control the Pattern of its Expression

BACKGROUND There is emerging data suggesting that the non-coding RNA (microRNA 193a or miR-193a) plays key roles in different types of cancers. OBJECTIVE This review aims to investigate the functional significance of miR-193a in different cancers according to the information of literature. METHOD All the literature concerning miR-193a in cancer in PubMed are analysed. RESULTS Several studies proved the association of miR-193a expression patterns with cancers stages, grades, response to the chemotherapy and even patient survival. Also, miR-193a can be used to differentiate some types of cancer. In cancer, miR-193a can act as a tumour suppressor gene or as an oncogene. Till now, several genetic factors (MAX, RXR α, XB130, P63, P73, AEG-1, HIFs, EGFR, Drosha, DGCR8, Dicer) and epigenetic factors (DNA methylation and long non-coding RNAs) were predicted to control miR-193a expression. They have fundamental effects on its biological behaviour in different types of cancers. CONCLUSION miR-193a has significant roles in cancer and can be targeted in the future for cancer therapy by better understanding of the factors that control its biological behaviour.

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.

Abstract 183: Inhibition of BRAF kinase alone in BRAF V600E-mutated undifferentiated thyroid carcinoma results in no growth arrest

Background: Undifferentiated thyroid carcinoma is the most aggressive type of thyroid carcinoma. It is refractory to most of the conventional therapies and its median overall survival is less than six months. Mutation in BRAF gene was found around 25% of undifferentiated thyroid carcinoma. Mutated BRAF particularly the V600E mutated BRAF auto-activates the BRAF-MEK proliferation-survival cell signalling pathway. The objectives of our current study were to inhibit the BRAF kinase and investigate its effect on undifferentiated thyroid carcinoma proliferation. Materials and methods: 8050C, a homozygous BRAF V600E mutate undifferentiated thyroid carcinoma cell line, was used to develop anti-BRAF shRNA stable cell line. Quantitative real time polymerase chain reaction (qPCR) and western blot were performed for ratifying the BRAF inhibition efficiency of stable cell line. Proliferation and colony formation assay were done to see the effect of BRAF inhibition on undifferentiated thyroid carcinoma proliferation. In addition, impact of BRAF inhibition on proliferation cell signalling pathways were checked through western blot analysis. Results: Efficiency of BRAF knockdown was around 87% which resulted in approximately 83% inhibition of BRAF kinase (p Conclusions: Inhibition of BRAF kinase reduced cell proliferation initially, but this proliferation inhibition did not lead to cell growth arrest at the end. Increased AKT kinase activity counteracted the effect of BRAF kinase inhibition as AKT kinase has the ability to contribute in overall cell proliferation. Therefore, both kinases need to be inhibited simultaneously for cell growth arrest in undifferentiated thyroid carcinoma. Citation Format: Md. Atiqur Rahman, Ali Salajegheh, Haleh Vosgha, Hamidreza Maroof, Robert A. Smith, Alfred KY Lam. Inhibition of BRAF kinase alone in BRAF V600E-mutated undifferentiated thyroid carcinoma results in no growth arrest. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 183.

Abstract 3393: miR-34b: A regulator of VEGF-A, Notch1 and Bcl-2 in thyroid carcinoma

Background: Thyroid cancer is the most common endocrine malignancy accounting for >91% of endocrine malignancy and 1.8% of all recently distinguished cancer reports, with an incidence continuing to rise globally in recent decades and current treatment strategies are not potent enough for patients. Therefore, novel and more effective treatment are outstandingly required. New strategy for treatment is target therapy, which not only hold cancer-specific expression but also limits side effects. miR-34b as part of p53 tumor suppressor network, plays crucial role in many physiological and pathological processes including cancer initiation, tumor progression and cancer angiogenesis process. Objectives: In this study, we evaluate the functional roles of miR-34b in 2 thyroid cancer cell lines for its potential in modulating angiogenesis and proliferation in thyroid malignancies. Methods: Expression levels of miR-34b was determined in metastasizing human papillary thyroid carcinoma (B-CPAP) and human undifferentiated thyroid carcinomas (MB-1) cell lines by qPCR. Exogenous miR-34b was transfected to thyroid cancer cell lines to investigate its effect on predominant genes involved in angiogenesis, cell cycle and apoptosis regulation including VEGF-A and Bcl-2 and Notch1. Confocal laser scanning microscopy (CLSM) and western blot techniques were performed to illustrate the protein expression changes. To demonstrate the perturbation of cell cycle and apoptosis pathways through exogenous miR-34b, fluorescence-activated cell sorting (FACS) was performed. Results: Significant underexpression of miR-34b was established in B-CPAP and MB-1 cell lines while outstanding overexpression of VEGF-A, Bcl-2 and Notch1 were detected. After transfection with miR-34b, significant drop in concentration of VEGF-A, Bcl-2 and Notch1 were noticed compared with controls in CLSM and western blot analysis (p Cell cycle analysis demonstrated that 48 hours after ectopic induction of miR-34b, significant impairment in proliferation of B-CPAP and MB-1 was induced through arresting cancer cell proliferation in G0-G1 phase (14.27% ±3.50 for B-CPAP and 13.61% ±0.16 for MB-1) (p Conclusion: These results pave the way that miR-34b may be involved in balancing tumour angiogenesis, cell proliferation and apoptosis in thyroid cancer. These could potentially occur via direct modulation of downstream targets such as VEGF-A, Bcl-2 and Notch1 for their role in those events. The future of this research will investigate the regulatory role of miR-34b in Papillary and undifferentiated thyroid carcinoma through larger scale in vivo and clinical trial studies which shall potentially improve the clinical presentation of aggressive malignancies. Citation Format: Hamidreza Maroof, Ali Salajegheh, Alfred KY Lam. miR-34b: A regulator of VEGF-A, Notch1 and Bcl-2 in thyroid carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3393.

Abstract 561: A natural copy number variant in the GAEC1 oncogene is associated with breast cancer susceptibility

The recently discovered GAEC1 oncogene, located at 7q22, has been found to be amplified in esophageal and colorectal cancers and is associated with altered clinical characteristics in those diseases. Amplification studies in these cancers showed variation in copy number in normal tissues, indicating a potential natural copy number variant (CNV) of GAEC1. In this research, the presence of such a CNV and its association with sporadic breast cancer development was tested in a population of 190 Caucasian women with breast cancer and a population of age, sex and ethnicity matched controls. Data produced by qPCR confirmed the presence of a CNV for GAEC1 in both cancer and control populations, with three common copy number types and several uncommon values, representing up to a 64-fold difference in GAEC1 copies. In addition, Chi-square analysis of copy number types indicated that increased GAEC1 copy number is associated with cancer development. This includes analysis using only the common copy number types (p=0.05) as well as considering all genotypes on a High/Low gene dosage basis (p=0.03). Overall risk for breast cancer was found to be slightly increased in high dose individuals (OR 1.552 (95% CI 1.035-2.338)). These results indicate that GAEC1 may be an important gene in the development of breast cancer, and that examination of its copy number in patient DNA may be a useful addition to genetic risk markers in sporadic cases. Citation Format: Vinod Gopalan, Robert A. Smith, Suja Pillai, Lyn R. Griffiths, Alfred K-Y Lam. A natural copy number variant in the GAEC1 oncogene is associated with breast cancer susceptibility. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 561. doi:10.1158/1538-7445.AM2014-561

Abstract 381: VEGF-A and VEGF-C: potential coregulators in angiogenic event in thyroid cancer.

Vascular endothelial growth factor (VEGF) promotes growth of blood or lymphatic vessels, termed angiogenesis and lymphangiogenesis. These share several basal mechanisms, including cross-talk between VEGF-A, C and their receptors, which may be important in the development of malignancies. Study aimed to identify relationships between VEGF-A, VEGF-C and their impact in angiogenesis and metastases in thyroid cancers.We studied VEGF-A and VEGF-C mRNA expression in 123 thyroid cancers, 40 lymph node metastases and 7 non-malignant thyroid control tissues using qPCR. Protein expressions for both VEGFs were determined by immunohistochemistry.VEGF-A and VEGF-C mRNA over-expression was noted in 51% (n=62) and 27% (n=33) of the thyroid cancers respectively. VEGF-A and VEGF-C protein over-expression was also identified in 70% (n=86) and 62% (n=76) of the thyroid cancers. ANOVA showed significant expression differences for VEGF-A and VEGF-C in subtypes of thyroid carcinoma, lymph node metastasis and normal controls (P=1x10-6 and 1x10-5 respectively). VEGF-A mRNA was significantly higher in cancers with lymph node metastases compared to non-metastatic cancers (p=0.001) however, most metastatic tumors under-expressed VEGF-C (p=0.0002). Similarly, VEGF-A protein expression was higher in metastases and VEGF-C protein was higher in non-metastatic lesions, though not significantly. Finally, we used linear regression analysis to test previous proposals about physiological interactions of VEGF-A and VEGF-C expression. Our model indicated that in metastatic cancers, 32% of the expression of VEGF-A could be predicted by a positive influence of VEGF-C production (model p=0.0002). These findings add weight to previous hypotheses concerning VEGF-A and C interaction. Early lymphangiogenic events were proposed to regulate angiogenesis, and changes to VEGF expression in this cancer population bear this out, with VEGF-C in early tumors giving way to VEGF-A in metastases. The commonality of local metastasis in thyroid cancer makes it a useful model to further examine mechanisms of angiogenesis and lymphangiogenesis in cancer metastasis. Keywords: thyroid cancer, VEGF-A, VEGF-C Citation Format: Ali Salajegheh, Robert A. Smith, Sahar Pakneshan, Vinod Gopalan, Alfred KY Lam. VEGF-A and VEGF-C: potential coregulators in angiogenic event in thyroid cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 381. doi:10.1158/1538-7445.AM2013-381