International Conference on Cancer Genetics and Epigenetics October 26-28 2017 Osaka,Japan

Kiyoon Kim has completed her PhD and Postdoctoral studies from School of Medicine, Kyung Hee University. She is a Research Professor of Department of Biochemistry and Molecular Biology.

Despite considerable progress in understanding the function of peroxiredoxin (Prx) in cancer, its expression patterns have not been extensively studied in response to cervical carcinogenesis. We evaluated the expression of Prx isoforms in normal tissue, cervical intraepithelial neoplasia (CIN1, CIN2 and CIN3) and cervical cancer. We found strong pattern of increased Prx II and III immunostaining with increasing severity of the lesion. No difference in staining intensity by grade of lesion was observed for Prx I and IV. Therefore, we conclude that Prx II and III are up-regulated in response to the development of cervical cancer.

Suyun Jeong has completed her Master’s degree from Kyunghee University in Seoul, South Korea. Presently, she is studying about viruses and cancer.

The study was performed to investigate the anti-cancer effect of novel compounds in the liver cancer. We performed with many synthetic compounds which are various forms of anti-cancers. We had screened test anti-cancer drugs and selected compatible two compounds. In the study, the anticancer effect and apoptotic molecular mechanism of two compounds were investigated on the human hepatocellular carcinoma cell, HepG2. The anti-proliferation effects of compounds on the HepG2 cell were analyzed using MTS. We showed the expression levels of anti-apoptotic (Bcl-2) proteins and apoptosisassociated proteins (Bax, caspase-3, PARP) using western blot. The results indicated that two compounds effectively inhibit cell proliferation and promote apoptosis and two compounds down-regulated the level of Bcl-2 protein and up-regulated the activity of Bax, cleaved-Parp, cleaved-Casepase-3 proteins. Therefore, two compounds induce apoptosis of hepG2 through the change of the apoptosis-associated proteins.

Young-Seok Lee has completed his Master's degree at Kyunghee University, Seoul, South Korea. Presently, he is a Doctoral student, studying about liver cancer therapy using newly-synthesized compound and protein proteasomal-degradation.

Our lab synthesizing new compounds, compound A, based on natural extracts. Compound A was modified by some residues of natural extracts. Compound A has lower side effects than well-known anticancer agents and has a higher cancer therapeutic effect than from backbone natural extracts used in the production of Compound A. Our experimental results that cell viability was decreased in dose-dependent manner via the MTS assay. Compound A repressed cell viability of HepG2 cell line after 24 hours treatment with dose ranged from 25 μΜ to 100 μΜ. We also confirm the cell death marker through Western blotting and FACs analysis and investigated that compound A induces apoptosis. In western blot data, Compound A was treated for 24 hours, dose-dependent manner on HepG2 cells and the apoptosis marker protein Cleaved-Caspase 3, Cleaved- PARP was dose-dependently increased. In addition, the pro-apoptotic marker Bax increased and anti-apoptotic marker Bcl2 was decreased. In FACs data, HepG2 cells were exposed to Compound A for 24 hours, which resulted in an accumulation of cells in G2/Mphase. According our data, we expect that we develop promising therapeutic agents that are lower price than the well-known drugs for liver cancer and have lower side effects and higher effectiveness using Compound A.

Chien-Hung Shih is currently a PhD student at Institute of Molecular Medicine of National Tsing Hua University, Taiwan. He has his specialization in cell and molecular biology and epigenetics. His study focuses on investigating definite biomarkers and molecular mechanisms for oral cancer invasion and metastasis to find out potential therapeutic targets and strategies.

Current anti-epidermal growth factor receptor (EGFR) therapy for oral cancer does not provide satisfactory efficacy due to drug resistance or reduced EGFR level. As an alternative candidate target for therapy, here we identified an oncogene, ROS1, as an important driver for oral squamous cell carcinoma (OSCC) metastasis. Among tumors from 188 oral cancer patients, up-regulated ROS1 expression strongly correlated with metastasis to lung and lymph nodes. Mechanistic studies uncover that the activated ROS1 results from highly expressed ROS1 gene instead of gene rearrangement, a phenomenon distinct from other cancers. Our data further reveal a novel mechanism that reduces histone methyltransferase EZH2 leads to a lower trimethylation of histone H3 lysine 27 suppressive modification, relaxes chromatin, and promotes the accessibility of the transcription factor STAT1 to the enhancer and the intron regions of ROS1 target genes, CXCL1 and GLI1, for up-regulating their expressions. Down-regulation of ROS1 in highly invasive OSCC cells, nevertheless, reduces cell proliferation and inhibits metastasis to lung in the tail-vein injection and the oral cavity xenograft models. Our findings highlight ROS1 as a candidate biomarker and therapeutic target for OSCC. Finally, we demonstrate that co-targeting of ROS1 and EGFR could potentially offer an effective oral cancer therapy.

Ju Hee Kim is currently working as a Research Fellow in Seoul National University Hospital, Seoul, South Korea. She has completed her PhD in Life Science, Ewha Womans University and Post-Doctorate in Catholic-Harvard Wellman Photomedicine Center, Seoul, South Korea.

Calsequestrin (CASQ) is a Ca2+-binding protein localized in the endoplasmic/sarcoplasmic reticulum (ER/SR), an intracellular Ca2+ release and storage of muscle. CASQ2 forms a complex with Ryanodine Receptor-2 (RyR2) luminal calcium release channel and the intrinsic membrane proteins Triadin and junction in cardiac muscle. Ca2+ is a sequester and regulator of diverse cellular processes and specific Ca2+ channels play important roles in cell proliferation and invasiveness of breast cancers. To know the role of CASQ2 in breast cancer cells, we established CASQ2 over-expressing stable cells in Hs578T cells using retrovirus. CASQ2 over-expressing Hs578T cells showed higher level of migration and invasion rate compared to Hs578T, which indicated that overexpression of CASQ2 related with cellular functions. We also found that CASQ2 overexpression elevates extracellular signal-related kinase (ERK) expression. In epidermal growth factor (EGF) treated cells, CASQ2 over-expressing Hs578T had higher phosphorylated ERK compared to Hs578T. The results from this study show a possible cause of migration and invasiveness in breast cancer cells. Taken together, these findings demonstrate that CASQ2 could be a new therapeutic target for breast cancer.

Bok Sil Hong has completed her PhD in 2011 from Pohang University of Science and Technology (POSTECH). She is currently a Research Professor of Seoul National University Hospital, Center for Medical Innovation. Her main research field is about the significant roles of miRNAs on breast cancer progression. She has published more than 15 papers regarding the cancer biology and progression.

MicroRNAs (miRNA) are involved in crucial biological processes such as cell proliferation, differentiation and apoptosis and the dysregulation of miRNA has been demonstrated in initiation and progression of a variety of human malignancies, including breast cancer. Several studies suggested that that down-regulation of miR-204-5p is associated with poor prognosis and metastasis of breast cancer by regulating proliferation, apoptosis, migration, invasion of breast cancer cells in vitro, however, the underlying mechanisms of in vivo anti-tumor or anti-metastatic activity of miR-204-5p remains to be elucidated. Here, we identified differentially expressed miRNAs between primary breast tumors and normal adjacent tissues using small RNA sequencing. Among them, miR-204-5p was the most significantly down-regulated in all of 17 breast tumor tissues compared with corresponding normal pairs. To investigate the role of miR-204-5p on breast cancer progression, we first established stable breast cancer cell lines overexpressing miR-204-5p. MiR-204-5p suppressed tumor growth and metastasis in both syngeneic 4T1 murine allograft breast cancer model and MDA-MB-231 xenograft model via inhibiting proliferation, migration and invasion of breast cancer cells. With RNA-seq and the integrative analysis of TCGA data, we observed that there was significant inverse correlation between miR-204-5p expression and PI3K/Akt/mTOR signaling pathway. After analyzing down-regulated genes and target prediction in silico, we found that PIK3CB, a catalytic subunit of PI3K, was highly down-regulated in miR- 204-5p overexpressing cells and experimentally validated that PIK3CB was a direct target of miR-204-5p. Also, knockdown of PIK3CB impaired breast cancer cell proliferation and migration. Moreover, miR-204-5p decreased the mobilization and recruitment of CD11b+ myeloid-derived suppressor cells, which facilitated cancer cell metastasis through regulating mTOR signaling pathway, via down-regulating the expression of CCL20, VEGFA and CSF1. In conclusion, miR-204-5p inhibits the initial progression of breast cancer and metastasis by blocking proliferation, migration as well as invasion. These findings indicate that miR-204-5p may be a novel therapeutic strategy against breast cancer progression and metastasis.

Shu-Chun Chang is an Assistant Professor of Translational Medicine, Taipei Medical University, Taiwan. She is trained in extracellular biology and glycobiology. Previously, she has worked in cancer research in Imperial College London. She is a Post-Doctorate in the field of Immunology/Inflammation at National University of Singapore. She has published more than 10 papers in reputed journals. Her research interests are in chronic inflammation-associated tumors (e.g., colon cancer and breast cancer) and this has a profound impact in identifying new molecular therapies for autoimmune diseases, immunodeficiency and cancers.

Chronic inflammation-mediated cell death/survival is an important risk factor to cancer development. However, the link between chronic inflammation and tumorigenesis is still unclear. Elucidating this link is needed for early diagnosis and development of therapeutics. Previously, we reported that SAG (sensitive to apoptosis gene) is a key regulator between immuneoveractivation and pro-tumorigenesis. Here, by retrospectively studying human primary hepatocellular carcinoma (HCC) tissues, we showed that SAG is up-regulated in the early stage of HCC, in conjunction with the increase in ubiquitination of specific suicide/apoptosis factors such as SARM and Noxa. We envisage that SAG-UPS (ubiquitin proteasome system) degrades apoptosis factors, thus conferring anti-apoptosis and uncontrolled cell survival strategy, which promotes liver cancer. We found that up-regulated SAG in HCC plays a key pro-tumorigenic role by perturbing the fine balance of the ratio of proand anti- apoptotic factors, as indicated by the production of pro- and anti-tumorigenic cytokines. This favors and exacerbates the vicious cycle of tumorigenic microenvironment for the progression of hepatoma. Our findings clearly established the power of SAG-UPS as a cell death/survival decision link between chronic inflammation and tumorigenesis. We propose SAGUPS to be an early diagnostic marker for HCC, and a potential target for therapeutics development.

Viktor N Tomilov has graduated from EPFL, Lausanne, Switzerland and obtained his MS degree from Novosibirsk University, Russia. He is the Head of Bionformatics Department, Department of Mathematical Biology in SibEnzyme Ltd. He has published more than 20 papers in the various scientific fields (chemistry, genetics, genomics, computer sciences, etc.).

An aberrant methylation of the genomic regulatory regions may disrupt normal functioning of cells and often accompanies the human diseases including cancer. Thus, DNA methylation markers have significant diagnostic value and attract attention of many biomedical researchers. Unfortunately, the existing approaches for genome-wide methylation study are rather expensive and laborious. We have developed and successfully tested a novel method of the methylated sites mapping in the genomes. The method is based on a property of methyl-dependent site-specific DNA endonuclease GlaI to cleave DNA only at R(5mC)GY sites forming blunt-ended fragments. The following next generation sequencing of the fragments obtained after GlaI hydrolysis of DNA from Raji malignant cell line allows to reveal more than 2,617,000 positions of R(5mC)GY sites in genome. We have applied this approach to find differentially methylated RGCY sites in the genomes of malignant Raji and U-937, as well as non-malignant L-68 lung fibroblast cell lines. The positions of methylated RCGY sites in the genomes have been determined using pair-ended sequencing of GlaI fragments. A comparison of the obtained data has revealed significant differences in methylation of RGCY sites in CpG islands, putative regulatory regions and some repetitive DNA families between all three genomes. GO enrichment analysis of genes with highly methylated regulatory regions has shown the metabolic processes, which may be affected epigenetically in carcinogenesis. The new method allows determining positions of many modified cytosine bases in the genomes and may be a simple alternative to the existing methods of genome-wide methylation analysis.

Evgeny Dubinin is a Senior Researcher at SibEnzyme Ltd. Russia. The main area of company’s activity is research of new DNA enzymes and their application in modern life sciences. We are the discoverers of new type of enzymes – methyl-directed DNA endonucleases which are very promising for epigenetic studies.

An aberrant methylation of the genes regulatory regions is shown for many cancer diseases. The detection of these epigenetic biomarkers is one of the most promising diagnostic and prognostic tools. Such de novo DNA methylation in cancer cells is catalyzed by DNA methyltransferases DNMT3A and DNMT3B, which methylate RCGY sequences with formation of R(5mC)GY sites. Based on the methyl-directed DNA endonuclease GlaI, we developed a GLAD-PCR assay, which allows a quick and inexpensive determination of R(5mC)GY site in a selected position of human genome without bisulfite conversion. Recently, we applied GLAD PCR assay for development of the epigenetic test for colorectal cancer. In the present study, we applied GLAD-PCR assay to find out marker R(5mC)GY sites for detection of lung cancer (LC) which is one of the most frequent malignancies. The studied group of patients included forty LC patients who had a surgery. A total 65 surgical resection samples were studied including lung cancer tissues of varying degree of differentiation (n=40) and paired normal lung tissue controls (n=25). In the present work, we studied four R(5mC)GY sites in regulation regions of HOXA5, LHX6, RASSF1 and RARB genes to identify aberrantly methylated RCGY sites. The analysis of RCGY sites methylation with GLAD-PCR assay demonstrated a relatively good prognostic potential of LC detection for RCGY sites in regulation regions of LHX6 and RASSF1 genes. We believe that these RCGY sites may be used for LC determination by GLAD PCR assay of DNA samples from blood and sputum.

Hee Jun Cho is a Senior Researcher with the Immunotherapy Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, South Korea.

Eph receptors and their corresponding ephrin ligands have been associated with regulating cell-cell adhesion and motility and thus play a critical role in various biological processes including tissue morphogenesis and homeostasis, as well as pathogenesis of several diseases. Aberrant regulation of Epinephrin signaling pathways is implicated in tumor progression of various human cancers. Here, we show that a Rho family GTPase regulator, Rho guanine nucleotide dissociation inhibitor-1 (RhoGDI1), can interact with ephrinB1 and this interaction is enhanced upon binding the extracellular domain of the cognate EphB2 receptor. Deletion mutagenesis revealed that amino acids 327~334 of the ephrinB1 intracellular domain are critical for the interaction with RhoGDI1. Stimulation with an EphB2 extracellular domain-Fc fusion protein (EphB2-Fc) induces RhoA activation and enhances the motility as well as invasiveness of wildtype-ephrinB1 expressing cells. These Eph-Fc-induced effects were markedly diminished in cells expressing the mutant ephrinB1 construct (Δ327-334) that is ineffective at interacting with RhoGDI1. Furthermore, ephrinB1 depletion by siRNA suppresses EphB2-Fc-induced RhoA activation and reduces motility and invasiveness of the SW480 and Hs578T human cancer cell lines. Our study connects the interaction between RhoGDI1 and ephrinB1 to the promotion of cancer cell behavior associated with tumor progression. This interaction may represent a therapeutic target in cancers that express ephrinB1.