University of Franca, Brazil
The Styrax camporum Pohl, commonly found in Brazil, is used for therapeutic purposes by folk medicine to treat ulcers, respiratory diseases and others. Phytochemical studies have demonstrated the presence of the benzofuran neolignans, egonol (EG) and homoegonol (HE), in all Styrax species. These lignans are therefore considered phytochemical markers for the quality control of Styrax extracts. These compounds possess some biological activities of clinical interest, such as antibacterial, antifungal and cytotoxic to tumor cells. Previous studies conducted by our group with Styrax camporum extract demonstrated its modulating effect on DNA damage induced by different mutagens. Thus, the aim of the study was to investigate whether the EG and HE are responsible by the modulating activity observed for the S. camporum extract. The study was carried out in Chinese hamster lung fibroblasts (V79 cells) using the micronucleus assay. The cultures were treated with EG (0.26 µg/mL), HE (0.017 µg/mL) alone or combined. In addition, the same concentrations were simultaneously treated with methyl methanesulfonate (MMS, 44 µg/mL), hydrogen peroxide (H2O2, 3.5 µg/mL), camptothecin (CPT 43 µg/mL) and etoposide (VP16 1 µg/mL). We also included a negative (without treatment), a solvent (dimethylsulfoxide; DMSO 0.05%) and positive control groups. The cultures treated with EG plus HE showed genotoxicity. The cultures treated with the EG and HE alone or combined with MMS or CPT no shown a significant effects. Regarding the combination with H2O2, the cultures treated with EG alone or combined with HE a significant increase on the micronuclei frequencies was observed when compared to cultures treated with H2O2 alone. Regarding the combination with VP16, the EG and HE alone or combined led to a reduction in the frequency of micronuclei induced by VP16. This study allowed us to better understand the mechanism of action of the S. camporum and chemical markers and suggest modulation depends on the type of mutagen used.
Financial Support: São Paulo Research Foundation (FAPESP, Brazil, grant # 2013/13903-9).
Denise Crispim Tavares is a Professor and researcher at the University of Franca, Franca, Brazil. She has experience in Genetics, with emphasis on Mutagenesis, acting on the following topics: assessment of genotoxic activity, antigenotoxic, antiproliferative and anticarcinogenic herbal extracts and isolated substances and synthetic.
Department of Pharmacology, School of Medicine, Chungnam National University, Korea
In this study, we investigated whether the flavonoids, wogonin and apigenin, significantly affect MUC5AC mucin gene expression and production in human mucoepidermoid carcinoma cells. Confluent NCI-H292 cells were pretreated with wogonin or apigenin for 30 min and then stimulated with tumor necrosis factor-α (TNF-α) for 24 h or the indicated periods. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. We found that incubation of NCI-H292 cells with wogonin or apigenin significantly inhibited mucin production and down-regulated MUC5AC gene expression induced by TNF- α, in a dose-dependent fashion. To elucidate the action mechanism of wogonin or apigenin, effect of each compound on TNF-a-induced NF-kB signaling pathway was investigated by western blot analysis. Wogonin inhibited NF-kB activation induced by TNF-a. Inhibition of IKK by wogonin led to the suppression of IkB phosphorylation and degradation, p65 nuclear translocation and NF-kB-regulated gene expression. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. Wogonin also inhibited the gene products involved in cell survival (Bcl-2) and proliferation (cyclooxygenase-2). Apigenin inhibited NF-κB activation induced by TNF-α. Inhibition of inhibitory kappa B kinase (IKK) by apigenin led to the suppression of inhibitory kappa B alpha (IκBα) phosphorylation and degradation, p65 nuclear translocation. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1). These results suggest that wogonin and apigenin inhibit the NF-kB signaling pathway, which may explain their roles in the inhibition of MUC5AC mucin gene expression and production from human mucoepidermoid carcinoma cells.
Dr. Choong Jae Lee is a Full Professor of Medical Pharmacology at the School of Medicine, Chungnam National University in Daejeon, South Korea. He obtained his PhD in Cellular and Molecular Pharmacology from the College of Pharmacy, Seoul National University in 1997. Dr. Lee did a postdoctoral training at Seoul National University where he carried out studies in pulmonary and anticancer pharmacology focusing the development of candidates for novel therapeutics. He worked as a research fellow at the School of Pharmacy, University of Maryland at Baltimore under advice of Dr. K. Chul Kim, in 1995. He joined Chungnam National University in 2002 as an Assistant Professor and is continuing his research in anticancer pharmacology with emphasis on elucidation of mechanism and action of potential chemopreventive natural products. He spent his sabbatical year at the University of California Santa Barbara studying cancer biology with the renowned professor Dr. Leslie Wilson, in 2008. Dr. Lee published over 40 research publications, majorly dealing with bioactive natural products, in top peer-reviewed scientific journals.
1Instituto de Química de São Carlos, Universidade de São Paulo, Brazil
2Departamento de Química, Universidade Federal de São Carlos, Brazil
A factor that limits the efficiency of most photosensitizers (PS) used in PDT is their tendency to self-aggregate due to the strong attractive interactions between π-systems of high conjugated molecules. Therefore, recent studies have focused on the development and efficacy of new photosensitizers. Chlorin derivatives present high absorption around 650-670 nm, strong ability to generate ROS and low cytotoxicity in the absence of light. In this study the synthesis of two chlorin through the Diels–Alder reaction of protoporphyrin IX dimethyl ester with an activated dienophile (1-(2-hydroxyethyl)-1H-pyrrole-2,5-dione) were performed. These new potential photosensitizers presented a special “L-shape” structure, and were sterically prevented from self-aggregation in solution. Singlet oxygen quantum yield of chlorin CHL-OH-A and CHL-OH-B were determined as ΦΔ= 0.49±0.04 and ΦΔ=0.43±0.02, respectively, when compared with methylene blue (ΦΔ= 0.52). Photo bleaching studies showed low photodegradation (~10%) after irradiation at the Soret band for 10 min at 50mW. The Log P of the CHL-OH-A and CHL-OH-B were also measured as 1.20±0.04 and 1.17±0.03, respectively, thus revealing these compounds as amphiphilic molecules. The cell viability studies with neutral red in tumoral Hep-2 cells after irradiation at 660 nm, showed IC50 of 7.26±0.77(A) and 6.05±0.73nmolL-1(B) at doses 3 Jcm-2 while at 6 Jcm-2 were 3.78±0.33 and 3.33±0.24nmolL-1 respectively. Moreover, fluorescence microscopy images using ethidium bromide and acridine orange showed a high percentage of apoptotic cells (90%) at the low concentration (10nmolL-1), suggesting these new compounds as potential candidates to photosensitizers. Acknowledgments: FAPESP and CAPES.
Janice Rodrigues Perussi is a Professor and researcher at the University of São Paulo, Brazil. She has experience with photosensitizers on the following topics: physical-chemical properties, photodynamic properties, cytotoxicity, type of cell death, intracellular accumulation, etc.
1Baylor Scott & White Memorial Hospital, USA
2University of Arizona College of Medicine, USA
Background: Breast cancer patients with diabetes have been shown to have an increased risk of overall mortality. Metformin use has been associated with improved survival. Still, there are other known risk factors such as stage, age, and pathology which influence breast cancer outcomes. The purpose of this study is to identify prognostic factors in breast cancer patients with and without diabetes.
Methods: We retrospectively reviewed a cohort of patients from a single institution (n=394) that received standardized, multi-disciplinary care for their breast cancer. Overall survival (OS) and recurrence free survival (RFS) were estimated by the Kaplan-Meier method. Cox proportional hazard regression models were utilized to determine different covariates on the risk of death and recurrence for diabetic and non-diabetic patients. Metformin use, age, surgery type, receptor status, BMI, and pathological features were also analyzed.
Results: Median follow-up for living and deceased patients was 9 and 5 years, respectively. 337 patients were identified as non-diabetic and 57 patients were diabetic. The two groups did not differ significantly in age, race, receptor status, surgery type, number of nodes, LVSI, tumor stage, and overall stage. The diabetic group had significantly more grade 2 and 3 histology and higher BMI. There was no difference in OS or RFS between non diabetic and diabetic patients (p= 0.879 and p=0.065 respectively). Metformin usage was not found to affect overall survival or recurrence free survival.
On multivariate analysis, significant predictors for OS included age, surgery type, ratio of positive nodes, and ECE. Significant predictors of RFS were surgery type, positive nodes, ECE, and grade. For every percent increase in ratio of positive nodes, the risk of death was increased by 1%. Those with ECE had a 3.31 fold increased risk of death. Patients with grade 3 were twice as likely to have a recurrence verses grade 2. For every 1 year increase in age, the risk of death increased by 1.9%.
On univariate analysis, underweight patients (BMI<18.5) had a 28.5% increased risk of recurrence and obese patients (BMI > 30) had a 40% increased risk of recurrence verses normal weight patients.
Conclusions: Unlike previous studies, we found no difference in OS in breast cancer patients with and without diabetes, as well as no difference with and without metformin use. Given the worse outcomes seen with age, BMI, ECE, grade 3, ratio of positive nodes (%), we recommend that these features be utilized as prognostic determinants to guide treatment decisions when applicable.
Divya Arora graduated with a Bachelors of Science in Nutrition at the University of Florida. She obtained a Masters of Biomedical Science at Tufts University. She completed by medical school at the University of South Florida in Tampa. She is now a PGY4 resident at Baylor Scott and White in Radiation Oncology. Her research interests include stereotactic radiosurgery and breast cancer.
1Gastroenterologie II, Klinikum rechts der Isar, TU München, Germany
2Research Unit Protein Science, Helmholtz Zentrum München, Germany
3Department of Surgery, Klinikum rechts der Isar, TU München, Germany
An organoid culture is a three dimensional (3D) cell culture that resembles an organ. Recently, organoid cultures were established from different organs such as small intestine, colon, stomach and pancreas; however those organoid cultures are exclusively composed of epithelial cells, thus missing the stromal niche. Here we aimed at the: 1) establishment of the organoid co-culture system to study stromal-epithelial cross-talk in the intestinal niche, 2) investigation whether fibroblasts could contribute to the early stages of tumorigenesis.
The niche was reconstructed by combining murine small intestinal crypts together with mesenchymal cells in 3D culture system. As mesenchymal cells the following cells were used: intestinal fibroblasts, carcinoma associated fibroblasts (CAFs), fetal esophageal fibroblasts and cardia fibroblasts. Mesenchymal-epithelial cross-talk was analyzed by phase contrast microscopy, Ki-67 and PAS staining, microarray analysis, mass spectrometry, and inhibitor studies.
In the co-culture, ~50% of crypts were growing as spheroids that were positive for Ki-67. Surprisingly, no differences between CAFs and other mesenchymal cells could be observed. By morphology and decreased number of PAS positive cells, the spheroids resembled intestinal organoids from Apc+/1638N mouse tumors. Transcriptomic analysis of crypts from the co-culture further confirmed that fibroblasts induce proliferation and do not promote differentiation in the crypts ex vivo. Experiments with different composition of culture medium, showed that spheroid formation was independent of R-Spondin. Moreover, pre-treatment of fibroblasts with IWP-2 (inhibitor of Wnt secretion) did not abrogate the spheroid formation. Indirect co-culture and conditioned media experiments revealed that spheroid induction was mediated by soluble factors. Mass spectrometric analysis of the secretome from the co-culture suggested the involvement of extracellular matrix-receptor pathway and focal adhesion pathway.
Taken together, our studies demonstrate that the microenvironment shapes intestinal crypts in a similar way as cell-autonomous genetic alterations and point out to the potential role of fibroblasts during tumor initiation.
The research interests of Agnieszka Pastuła include tumor microenvironment and three dimensional cell biology. Agnieszka Pastuła completed her Master degree in biology with honors at the Jagiellonian University in Cracow in 2009. She gained experience in research by doing multiple internships and working as a research assistant in Poland, the Netherlands, and Germany. Since 2011 she has been doing doctorate at the Klinikum rechts der Isar TUM, where she received the Forschungsförderung der Dr.-Ing. Leonard Lorenz-Stiftung and Laura Bassi Award. She is a reviewer of the Bio-Protocol, and a member of the American Association for Cancer Research and Deutsche Krebsgesellschaft.
1Johns Hopkins University School of Nursing, USA
3The Memory Clinic in Neuropsychiatry, Sheppard Pratt Health Systems; Department of Psychiatry and Behavioral Sciences, Johns Hopkins University, USA
4Research and Development Center, Institute of Health and Caring Sciences, Skaraborg Hospital, Sweden
5Swedish Society for Nursing, Karolinska Institution Division of Nursing, Sweden
6Acute and Chronic Care, Johns Hopkins University School of Nursing, USA
Little is known about the effects of burdensome symptoms during chemotherapy treatment in African-American women. This study explored the symptom burden occurring during chemotherapy treatment and how these symptoms impacted functional well-being and quality of life (QOL). A sample of 30 African American women with breast cancer (BC) completed a battery of questionnaires that were used to collect the data at baseline, midpoint, and at the completion of chemotherapy. There were significant differences in the severity of symptoms for worse pain, pain interference with activities of daily living (ADLs), present fatigue and history of fatigue, present nausea and history of nausea and insomnia as well as lower intensity of QOL measures over the course of chemotherapy treatment. All symptoms had greater intensity at midpoint and completion than at baseline. Worst pain had a significant negative effect on functionalwell-being. Both pain and depression each had significant negative effects on QOL.
Iye Kanu is a research nurse at Johns Hopkins University, School of Nursing and presently working on research projects aimed at reducing health disparities among minority women in oncology and perinatal settings.
1School of Medicine, Chung Shan Medical University, Taiwan
2Department of Medical Education, Cathay General Hospital, Taiwan
PD-L1 has been widely demonstrated to contribute to failed antitumor immunity. Blockade of PD-L1 with monoclonal antibody could modulate the tumor immune environment to augment immunotherapy. PD-L1 expression is also detected in several types of cancer and is associated with poor prognosis. However, the prognostic role of PD-L1 inoral squamous cell carcinoma (OSCC) is still controversial. Our aim was to determine the role of PD-L1 in the prognosis of OSCC patients to identify its potential therapeutic relevance. PD-L1 immunoreactivity was analyzed by immunohistochemistry in 305 cancer specimens from primary OSCC patients. The medium follow-up time after surgery was 3.8 years (range from 0.1 to 11.1 years). The prognostic value of PD-L1 on overall survival was determined by Kaplan-Meier analysis and Cox proportional hazard models. Higher PD-L1 expression is more likely in tumor tissues of female than male OSCC patients (P=0.0062). Patients with distant metastasis also had high PD-L1 expression (P=0.0103). Multivariate analysis identified high PD-L1 expression as an independent risk factor in males and smokers (males: hazard ratio =1.556, P=0.0077; smokers: hazard ratio =2.058, P=0.0004). We suggest that PD-L1 expression, determined by IHC staining, could be an independent prognostic marker for OSCC patients who are male or who have a smoking habit.
Mr. Shih-Chen Tsai is from the School of Medicine, Chung Shan Medical University, Taiwan and Department of Medical Education, Cathay General Hospital, Taipei, Taiwan
Department of Applied Biology & Chemical Technology, The Hong Kong Polytechnic University, Hong Kong
Background: Quinoline belongs to an important class of heterocyclic alkaloids. Anti-tumor activity of quinoline derivatives have been intensively investigated for the purpose of identifying novel anti-cancer agents. The purpose of this study is to evaluate the anti-tumor activities of a novel quinoline derivative 91b and identify the downstream regulatory targets.
Method: A novel quinoline derivative 91b was synthesized using the asymmetric hydrogenation reaction. MTS cytotoxicity assay was performed on the human esophageal cancer cell lines KYSE150 and KYSE450. The subcutaneous tumors of KYSE150 in athymic nude mice were also treated with 91b1 (10mg/kg/day as normal dose, and 50 mg/kg/day as high dose) via the intraperitoneal route for 1 month. The body weight and tumor volume were recorded regularly, and serums were collected to identity the changes of biochemical markers. SD rats were treated with 500mg/kg/day 91b for 7 days to examine the chronic toxicity. cDNA microarray analysis on KYSE150 cells treated with 91b was also performed to identify the downstream regulatory targets at cDNA level. One of the most down-regulated gene, Lumican, was cloned and transfected into NIH 3T3 cells for subcutaneous injection into athymic nude mice to determine its tumor transforming capacity.
Results: The MTS50 values ranged from 0.375 to 5.91 mg/ml for KYSE150 and 0.572 to 9.06 mg /ml for KYSE450 cell lines. Tumor volume of the treated group showed significant reduce (p=0.007) against the vehicle control group and the animal body weights increased significantly. ALB and ALP level were decreased compare to blank control group (ALB from 27.6 to 26.6, and ALP from 57.00 to 56.40). For the chronic toxicity test, the compound-treated group all survived. From the cDNA microarray analysis, Lumican is one of the most down-regulated genes in the 91b1 treated KYSE150 cells compared with the vehicle controls.
Conclusions: Compound 91b is a promising anti-tumor agent with high activity and low toxicity. Lumican is the most downregulated target. The tumor transforming capacity of lumican is under investigation using the in vitro and in vivo approaches.
Yuanyuan ZHOU is currently a PhD candidate at The Hong Kong Polytechnic University, majoring in applied biology and chemistry techniques. She completed her Masters at the China Pharmaceutical University, majoring in pharmacokinetics. She graduated with a bachelor of science also at the China Pharmaceutical University. Her research interest is on the characterization and mechanism of quinoline derivatives as novel anti-tumor agents.
1Hugh Adam Cancer Epidemiology Unit, Department of Preventive and Social Medicine, Dunedin School of Medicine, University of Otago, New Zealand
2Centre for International Health, Department of Preventive and Social Medicine, Dunedin School of Medicine, University of Otago, New Zealand
3Department of Obstetrics and Gynecology, KomfoAnokye Teaching Hospital, Ghana
Background: Globally, cervical cancer remains the fourth most common cancer. Despite the fact that cervical cancer is potentially preventable through screening and recently through vaccination, an estimated 266,000 women died worldwide of the disease in 2012. Incidence and mortality have reduced in most high-income countries mainly due to the introduction of cervical cancer screening. However, this is not the case in most low-income countries where approximately 85% of the disease occurs. In Ghana, cervical cancer is the most common cancer among women. The true incidence and outcomes of the disease in Ghana are not known.
Aims: To estimate the incidence and mortality rates of cervical cancer in Ghana.
Methods: Information on invasive cervical cancer cases diagnosed between 2010 to 2013 were collected from KomfoAnokye and Korle Bu Teaching hospitals through review of medical, pathology and computer records at the oncology unit and the obstetrics and gynecology department. These hospitals are the main referral centers in Ghana where patients with suspected malignancy are diagnosed and treated. Patients with cervical cancer receive diagnosis and treatment at these hospitals. Telephone interviews were also conducted with patients and relatives to gather further data. Data were recorded to a standardized study questionnaire and analyzed using summary statistics.
Results: A total of 1,725 women with cervical cancer were included in the study. Their ages ranged from 11-100 years with a mean of 56.9 years. Histology of primary tumor was the basis of diagnosis in 77.5% with clinical diagnosis accounting for 22.5%. For the 1,336 women with tumor grade available, moderately differentiated tumors accounted for 34.5%. Late stage at presentation was common. The highest number of cases was reported for women aged 50-54 years. As expected, the incidence of cervical cancer increased with age. However, the incidence rate was highest for the 75 to 79 year age group and began to decrease at older ages. The estimated crude incidence rate for the two regions combined (Greater Accra and Ashanti region) was 11.6 (95% CI: 11.0-12.2) per 100,000 person-years, and the ASR rate (World standard population) for incidence was 18.7(95% CI: 17.8-19.6). Highest rates of mortality were observed for women 75-79 of age. The crude mortality rate for the two regions combined was 5.5 (95% CI: 5.0-6.0) per 100,000 person-years, with an ASR of 7.8(95% CI: 7.1-8.5).
Conclusions: Late stage at diagnosis is common. The incidence and mortality rate of cervical cancer in Ghana increased with age. Our study suggests that considerable improvement in patient outcomes in low-income countries is needed through raising awareness and knowledge of the disease.
Funding source: The Directorʼs Cancer Research Trust of the Hugh Adam Cancer Epidemiology Unit provided funding for this research.
Yvonne Nartey is currently a PhD candidate at the University of Otago where she is specializing in cancer epidemiology. She completed her Masters at the University of Oxford, majoring in Global Health Science. She received a BSc in Nursing from the University of Ghana. During Yvonneʼs tenure as Masters student at the University of Oxford, she undertook a research with the Cancer Epidemiology Unit to study the incidence of cancer in Black African women as part of the Million Women Studies. Yvonne has since developed keen interest to work with the Ministry of Health in Ghana and other health institutions to address the various public health issues confronting the West African sub-region, especially cancer.
Chu Kochen Honors College, Zhejiang University, China
Eukaryotic translation initiation factors (eIFs) primarily form the translation initiation complex to start the synthesis of proteins. Aberrant expressions/subcellular distributions of many subunits of different eIFs have been shown to be associated with malignant phenotypes of cancer. Previously we found that eIF3g is up-regulated in multidrug resistant leukemia K562/ADR cells by proteomic approach. In this study, we analyzed the clinical significance of altered expression of eIF3g in human breast cancer, which showed strong association between high-level expression of eIF3g and lymph node metastasis. We further investigated the roles eIF3g plays on malignant phenotypes of breast cancer cell in vitro. Enforced expression of eIF3g in breast cancer cell Bcap 37 had no significant impact on the proliferation of the cells in vitro, but markedly enhanced the motility/invasiveness of the cancer cells. Knockdown of eIF3g in Bcap37 inhibited the proliferation of the cells but no influence on the survival of the cells. In conclusion, over-expression of eIF3g in breast cancer cells may contribute to a more aggressive phenotype.
(Sponsored by grant from National Natural Science Foundation of China 81172516)
Jingjia Ye obtained her PhD from Miyazaki University, Japan, majoring in oncology. She has worked in Cancer Institute, Zhejiang University and Clinical Research Center, the 2nd AffiliatedHospital, Zhejjiang University School of Medicine. Her research interest is on molecular mechanisms of tumor development and progression, early diagnosis for tumor micro metastasis and biotherapy of cancer. Now she works in Chu Kochen Honors College, Zhejiang University.
Department of Applied biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, China
Many bioactive compounds, both natural and synthetic molecules, are heterocyclic compounds. Recent studies have been reported that heterocycles, such as quinoline and indole, exhibit their promising anticancer effects. In present study, we examined the anticancer effects of some indole-type compounds as well as a series of benzenesulfonate quinolines were prepared and evaluated their potential anticancer activities against a panel of cancer cell lines (Hep3B, MCF-7 and SLMT-1). Among these compounds, compound QD019 demonstrated the best anticancer activity in all the selected cell lines with the MTS50 ranged from 0.078 to 0.625 µg/mL. According to the preliminary docking analysis using the Similarity Ensemble Approach (SEA), QD019 may potentially interact with the DNA binding site of the nuclear factor kappa beta p65 subunit which is critical for the proliferation of cancer cells. These findings provide a clue for us to further investigate it as a lead compound for its anticancer actions. Further structural optimization of the molecular skeleton and detailed investigation for the mechanisms of the anticancer actions of QD019 is in progress.
Po-Yee Chung is currently a research student at The Hong Kong Polytechnic University under the supervision of Dr. Johnny Cheuk-on Tang and Dr. Kim-Hung Lam. Her research focuses on drug design and synthesis of novel small organic molecules, particular in quinoline-type compounds, as anticancer agents.
Shanghai Jiaotong University School of Medicine, Shanghai Institute of Immunology, China
Ovarian cancer-associated antigen 66 (OVA66), also known as CML66 (GenBank Accession No. AF283301) was first identified in an ovarian carcinoma cDNA expression library and was shown to play a role in tumorigenesis. Here, we find that OVA66 influences tumorigenesis by regulating the type I insulin-like growth factor receptor (IGF-1R) signaling pathway. Stable knockdown of OVA66 in cancer cells attenuated phosphorylation of IGF-1R and ERK1/2-Hsp27; similarly, a higher level of p-IGF-1R and ERK1/2-Hsp27 signaling were also detected after OVA66 over expression in HO8910 cells. In vivo, knockdown of OVA66 both reduced tumor burden in nude mice and decreased phosphorylation of IGF-1R, ERK1/2, and hsp27. We blocked IGF-1R function both by siRNA and with the chemical inhibitor Linsitinib (OSI-906). By either method, tumorigenesis was inhibited regardless of OVA66 expression; thus, mechanistically, IGF-1R likely lies downstream of OVA66 in cancer cells. We also found that OVA66 regulates expression of MDM2; this attenuates ubiquitination of IGF-1R in response to IGF-1 stimulation and promotes active ERK1/2 signaling. Thus, we propose that combined over expression of OVA66 and MDM2 promotes oncogenesis by enhancing activation of the IGF-1R-ERK1/2 signaling pathway.
Mr. Yebin Xi is from Department of Immunology and Microbiology, Shanghai Jiaotong University School of Medicine. He graduated from Shanghai Second Medical University in 2001 majoring in medicine.
Shanghai Jiaotong University School of Medicine, Shanghai Institute of Immunology, China
Objective: To observe the expression of TAGLN gene in gastric cancer and its effect on the tumor invasion and migration.
Methods: Real-time PCR, Western blotting and immunohistochemistry were applied to examine the expression of TAGLN in gastric cancer tissues and cancer-adjacent tissues (40 cases), gastric cancer cell lines (7 cell lines) and carcinoma-associated fibroblasts (CAFs), respectively. The relationship between the TAGLN expression and the pathological characteristics of gastric cancer was also analyzed. To further evaluate the effect of fibroblasts expressing high TAGLN (such as CAFs) on the tumor metastasis, cell invasion and migration kits were used.
Results: The mRNA and protein expression of TAGLN in gastric cancer cel1 lines were much lower than those in tissues, especially the cell line MKN45. However, no significant difference was detected among gastric cancer cell lines (p>0.05). Compared with the cancer-adjacent tissues, the expression of TAGLN in gastric cancer tissues was higher (p<0.05). lmmunohistochemistry was also performed in our study and the TAGLN was found to be expressed mainly in stroma of gastric cancer, such as fibroblasts, endothelial cells of new vessels or myocyte in tumor-invaded muscu1aris mucosae. Furthermore, compared with that in lymph node free group (1.084±0.328), the TAGLN expression increased obviously in lymph node metastasis group (3.751±0.681). The cell invasion and migration examination in our study indicated that the CAFs expressing high TAGLN enhanced the invasion and migration ability of MKN45. Interestingly, when the TAGLN of CAFs was knocked down using siRNA, these cells may 1ost the ability to facilitate the invasion and migration of MKN45.
Conclusion: The TAGLN is overexpressed in the stroma of gastric cancer, and it may enhance the cell invasion and migration ability of gastric cancer.
Li Wang is a Lecturer in the Dept. of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, China. She did her PhD. In Immunology, Study on auto-irradiated T cells enhances anti-tumor immunity at Shanghai Jiao Tong University School of Medicine, China.
1Dept. of Medical Technology, International University of Health and Welfare, Japan
2Dept. of Obstetrics and Gynecology, Shinshu University School of Medicine, Japan
3Dept. of Laboratory Medicine, Shinshu University Hospital, Japan
4Dept. of Obstetrics and Gynecology, Osaka City University School of Medicine, Japan
5Dept. of Pathology, Keio University School of Medicine, Japan
6The University of Tokyo School of Medicine, Japan
7Dept. of Biology, Massachusetts Institute of Technology, USA
8Dept. of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, Japan
9Dept. of Obstetrics and Gynecology, Kyoto University Graduate School of Medicine, Japan
10Promoting Business using Advanced Technology, Japan Science and Technology Agency (JST), Japan
11The International Human Epigenome Consortium (IHEC) and CREST, Japan Science and Technology Agency (JST), Japan
Background: Although the majorities of smooth muscle neoplasm found in the uterus are benign, uterine leiomyosarcoma (LMS) is extremely malignant, with high rates of recurrence and metastasis. We earlier reported that mice with a homozygous deficiency for Lmp2/b1i, an interferon (IFN)-γ-inducible factor, spontaneously develop uterine LMS. The IFN-γ signaling pathway is important for control of tumor growth and invasion and has been implicated in several malignant tumors.
Aim: It is necessary to analyze risk factors associated with human uterine LMS, in order to establish a diagnostic biomarker and a clinical treatment method.
Methods and Results: In this study, experiments with mouse uterine tissues and human clinical materials revealed a defective LMP2/b1i expression in human uterine LMS that was traced to the IFN-γ signaling pathway and the specific effect of JAK-1 somatic mutations on the transcriptional activation of Lmp2/b1i gene. Furthermore, analysis of a human uterine LMS cell line and human clinical materials clarified the biological significance of LMP2/b1i in malignant myometrium transformation and tumor senescence, thus implicating LMP2/b1i as an anti-sarcomagenic candidate.
Conclusion: LMP2/b1i differential expression may be potential diagnostic biomarker for human uterine mesenchymal tumours, especially human uterine LMS. This role of LMP2/b1i as a tumour suppressor may lead to new diagnostic biomarker and therapeutic target molecule in human uterine LMS.
Dr. Takuma Hayashi is Professor at Dept. of Medical Technology, International University of Health and Welfare since 2016. He received his Doctorate from Inst. for Medical Science, University of Tokyo in 1994. He was clinical research training as a resident staff in National Cancer Center, Tokyo Japan for 3 years until 1994 and joined Whitehead Institute (WI)/Mass. Inst. of Tech.(M.I.T.) that year. He did postdoctoral training in the laboratory of Dr. Rick A. Young (Membership in the National Academy of Sciences, WI/M.I.T.), and also was a research member of USA Project of AIDS vaccine development (Project Leader: Dr. David Baltimore, Nobel Laureate, Cal.Tech.). After postdoctoral training, he got faculty position Lecture, Mass. General Hospital (MGH)/Harvard Medical School (HMS) in 1997. He has been studying the antigen presentation system by MHC class I with LMP2/b1i-deficient mice, under the cooperation of Dr. Susumu Tonegawa (Nobel Laureate, M.I.T.). He identifies diagnostic biomarkers, LMP2, Cyclin B1 and Cyclin E, for malignant tumour uterine leiomyosarcoma. Current research focus: molecular approach of tumorigenesis of uterine leiomyosarcoma.
Semey State Medical University, Kazakhstan
The goal: The identification of the presence and degree of endothelial dysfunction in patients with rectal cancer in the early and late periods after treatment. The study involved 47 patients with stage III clinical colorectal cancer (TNM), subjected to the combined therapy. Investigated: circulating endothelial cells, von Willebrand factor, metabolites of nitric oxide (NO); endothelium-dependent vasodilation.
Results: The excess of circulating endothelial cells over the control group - 4.25 times (p <0,001), excess WF - 2.11 times (p = 0.005). Revealed high levels of NO metabolites, differences with the control - 2.29 times (p = 0.005) reduction in the average value EDVD differences with the control - 1.92 times.
The excess of CE after surgery was 1.74 times (p = 0.01), compared to control excess - 7.42 times (p <0.001). WF increases, which reflected the endothelial damage in pathology and intervention, abuse aftercare - 2.52 (p = 0.003). Registered growth of NO metabolites. Differences with the control of postoperative 2.68 (p = 0.002). In the postoperative period the average value decreased EDVD differences with the control of 2.35 times (p = 0.004).
In the long term, 1 year after surgery, showed a trend reduction of the degree of dysfunctions, not reaching full normalization. Differences were found mainly in the number of circulating endothelial cells. The excess over the control of this indicator was 3.85 times (p = 0.023). At the same time, the average value of the indicator with respect to the state after the intervention decreased almost 2-fold (p = 0.027).
A significant excess (2-fold, p = 0.031) over the control group was determined by the content of von Willebrand factor. Marked differences were noted on the content of NO metabolites (2.09-fold, p = 0.025). Differences in the level EDVD decreased to 1.58 times (p> 0.05) due to a statistically significant increase relative to the level of the postoperative period.
Conclusion: rectal cancer the presence of endothelial dysfunction was observed before and after treatment. The greatest expression of disorders, is to increase the level of circulating endothelial cells, von Willebrand factor and reduction of vasodilatation, achieved after surgery and then gradually offset. With the development of recurrence and metastasis, the compensation does not occur, thereby increasing the risk of thrombotic complications.
Laura Pak graduated from high school in 2004 and in the same year entered to the Semey State Medical University in the specialty “General medicine”. In 2010 she graduated from the University and received a degree in “Medical doctor”. In the same year she was enrolled in the internship to the SSMU in 2011, at the end of the internship was certified “physician”. From 2011 to 2013 she held a residency at the Department of Oncology and Visual Diagnostics and received a certificate “Oncologist.” From 2013 till the present she has been studying in doctoral PhD, 3rd course in the SSMU, specialty is “Medicine”. The main directions of her training in doctoral studies: oncology, namely malignant tumors of the rectum.
Semey State Medical University, Kazakhstan
Introduction: Colorectal cancer (CRC) is the third most common cancer globally. It is also the fourth most common cause of cancer mortality worldwide, following lung, liver and stomach cancer. CRC incidence and mortality in Kazakhstan are relatively high but exact statistics appears to be lacking and trends over time are unclear.
Materials and Methods: The main sources of information for this research were the materials of the state colorectal cancer registration. We analyzed data for the study period of 10 years (2004-2013).
Results: The present study was therefore undertaken to retrospectively assess age standardized data for incidence and mortality were generated and compared across age groups, gender and year. It was determined that during that studies period 3,417(1) new cases of colorectal cancer were registered and 2,259(0,69) died of this pathology. The male to female ratios also didnʼt significantly vary over time but a trend for improvement of the mortality to incidence ratio was observed, especially for rectum. Whether this might be related to screening remains unclear. These preliminary data indicate that whereas CRC continues to be important, change in environmental factors are not having a great impact in East Kazakhstan.
Discussion: The present study showed that the average incidence of CRC in East Kazakhstan was around 24/100,000 with very little change over time, with again a relatively constant value of 15,9/100,000 for mortality. Comparison with other countries in the region is made difficult by the fact that our data are not age-adjusted. A large number of questions remain to be answered. For example, it is well known that ethnicity can play a role in determining risk and future work should focus on any differences between the main Russian and Kazakh populations. In conclusion, our data provide an initial survey of CRC in Kazakhstan, a Central Asia country with great geographical and ethnic variation.
Zhabagin Kuantkan Talgatovich, graduated in 2009 with a degree in general medicine. From 2009 to 2010 he held an internship in surgery. From 2010 to 2012 he passed residency in oncology. From 2013 year till 2016 year PhD student. Since 2011 is the assistant chair of the Semey State Medical University and is a medical oncologist at the Semey Oncology Center. Learning etiology, epidemiology, clinic of tumors of the gastrointestinal tract, malignant lymphoma, Hodgkinʼs disease, bone tumors and soft tissue.
1Pharmacology and Toxicology, Faculty of Pharmacy, Ain Shams University, Egypt
2Biology, School of Sciences and Engineering, The American University in Cairo, Egypt
Colorectal cancer (CRC) is the third common cause of cancer deaths. Chronic inflammation plays an important role in the pathogenesis of CRC. Cyclooxygenase-2 is overexpressed in several human cancers including CRC. Estrogen receptor-β (ER-β) is a subtype of estrogen receptors (ER) that possesses anti-inflammatory effects. It is the prevalent ER in normal colon mucosa and its level is significantly reduced in CRC. Caffeic acid phenethyl ester (CAPE) is an active component of honey bee propolis that possesses potent anti-inflammatory and selective estrogen receptor modulatory activities. The current work aims at investigating the potential modulatory effects of CAPE on the cytotoxicity of 5-Flourouracil (5-FU) in colon cancer cells and exploring the possible underlying mechanisms. Our data indicated that CAPE synergized the cytotoxicity of 5-FU in HT-29 cells as confirmed via Calcusyn synergy analyses. CAPE treatment significantly reduced cyclin D1 and increased p27 expressions compared to control. FACscan revealed an increased percentage of cells at pre-G phase with CAPE treatment. Bax/Bcl-2 ratio was significantly increased in the same group. CAPE increased the expression of ER-β and its downstream tumor suppressor FOXO1. Furthermore, CAPE treatment significantly reduced the level of COX-2, PGE2 and VEGF in HT-29 cells compared to control. In conclusion, CAPE synergized 5-FU cytotoxicity and antiproliferative activity in HT-29 cells. This can be, at least partly, attributed to CAPE pro-apoptotic effects, in addition to ER-β and COX-2 modulatory effects. Further investigations are needed in order to evaluate the potential merit of CAPE/5-FU combinations for the management of colorectal cancer in humans.
Dr. Mai F Tolba is an Assistant Professor of Pharmacology and Toxicology at Ain Shams University and The American University in Cairo. She is a post-doctoral member of the Society of Toxicology (SOT). Dr. Tolba is interested in investigating the role of estrogen signaling in cancer for which she has co-authored 3 publications. The cancer transforming role of catechol estrogens in prostate cells [PMID23685341]. The use of CAPE, a phytoestorgen, as a sensitizer for prostate cancer cells to taxanes [PMID23847086]. The effect of the obesity mediator leptin on estrogen metabolism and the proliferation of prostate cells [PMID25433128]
1Biotechnology Research Center, Pasteur Institute of Iran, Iran
2Department of Pharmacology-Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran
FHIT as a tumor suppressor executes a critical function in inhibiting p53 degradation by MDM2. Previous studies have revealed the interaction of FHIT with MDM2 or p53; however, there is no detailed study for interpreting the functional domains of FHIT involved in the tumor inhibition. In this study, we evaluated the various domains of FHIT owning the cytotoxic properties of the full length protein. Based on our previous in silico screening and MTT results, truncated forms of FHIT were selected and their cell cycle inhibiting properties were assessed in HT1080 human fibrosarcoma cell line. The functional analysis showed that these fragments can arrest cells in G2 phase of the cell cycle as indicated by flowcytometry.
These findings suggest FHIT functional domains as lead compounds for the discovery of future drug design and/or gene transfer for cancer therapy.
Keywords: Fragile histidine triad, HT1080, flowcytometry
Dr. Ameneh Eslamparast is an assistant professor of pharmaceutical biotechnology at Ardabil University of Medical Sciences. She obtained her Pharm. D. from the Shahid Beheshti University of Medical Sciences in 2005. She obtained her PhD in Pharmaceutical Biotechnology from the Pasteur Institute of Iran in 2014, where she carried out studies about FHIT anticanceric truncates. She joined Ardabil University of Medical Sciences in 2014 as an Assistant Professor and is continuing her research studies about anti-cancers with emphasis on peptides and plant extracts. Dr. Ameneh Eslamparast is a scientific writer with research publications in peer-reviewed scientific journals. Dr. Ameneh Eslamparast has received awards including the Ministry of health of Iran scholarship, for PhD course and Pasteur Institute of Iran award for cancer research.
1Saveetha Dental College, Saveetha University, India
2Department of Genetics, University of Madras, India
Background: Oral cancer is the eighth most prevalent cancer in the world but in India, it accounts for 30% of cancer deaths. In this study we focus on the possible role of Canonical Wnt signalling pathway in oral cancer in patients of South Indian origin. Although literature says that there are several components of this pathway altered in oral cancer, not many studies have been done on Indian population where the burden of this disease is on a high.
Objectives: To understand the role of WNT pathway genes in oral squamous cell carcinoma and to correlate the molecular findings with clinical behaviour.
Methods: A prospective study of 33 patients with oral cancer who reported to Saveetha University between July 2014 and January 2016 is done. Oral cancer tissues from surgically resected specimen of these patients, were processed for molecular study. Gene and protein expression: SFRP-1, WNT-3a, β-catenin, c-MYC and Cyclin-D are investigated in 33 oral cancer patients and 3 normal oral tissue samples using qRT-PCR and western blotting. Statistical analyses were performed using GraphPad Prism 6 (GraphPad software Inc., La Jolla, CA, USA) for validating gene expression. Data were analyzed by the Mann–Whitney test. All tests were two tailed and p < 0.05 was considered significant.
Results: We observed a high expression in all genes analyzed (SFRP-1, WNT-3a, β-catenin and Cyclin-D) by both real-time and western blot in the oral cancer tissues of these patients. Interestingly the mRNA levels of c-MYC in cancer tissues were lower compared to normal tissues, while c-MYC protein levels was found to be higher in cancer tissues. Continuous follow up of these patients are being done.
Conclusion: The decreasing expression levels of β catenin with progressing stages and increased levels of its target genes Cyclin D and C-myc can be useful markers and molecular targets for diagnostic and therapeutic intervention. Further validation is needed with IHC and epigenetic studies, which are ongoing.
Dr. Madhulaxmi Marimuthu has completed her Masters in Oral Surgery and is currently pursuing her PhD in Saveetha University, India. Her research interest is Oral Cancer, its molecular biology and its clinical implications.
Cancer Biology and Inflammatory Disorder Division, CSIR-Indian Institute of Chemical Biology, India
Cancer stem cells emerges as one of the most vital tumor initiation and maintenance system in almost all types of malignancies. Implicit is the role of CD44 in defining CSCs but CD24 is not well-explored in Oral Squamous Cell Carcinoma. Here we show that CD44highCD24low cells isolated from the oral cancer cell lines, not only express stem cell related genes but also exhibit Epithelialto-Mesenchymal transition (EMT) characteristics. Typical Cancer Stem Cell (CSC) phenotypes like increased colony formation, sphere forming ability, migration and invasion were also confirmed in CD44highCD24low cells. These cells also showed enhanced resistance to apoptosis inducing stimuli owing to their slow-cycling state and high expression of drug transporters. Dysregulation of microRNAs (miRNAs) are known to be implicated in tumor prognosis but whether it regulates stemness in OSCC remains unclear. We have shown miR-146a to be significantly up-regulated in CSCs derived from oral cancer cell lines and tumors. Enforced expression of miR-146a caused enhanced expression of CSC markers and clonogeneic potential. It was also linked with epithelial-mesenchymal transition (EMT) in addition to activated wnt signaling. Interestingly, miR-146a induces CSC traits by stabilising β-catenin that co-incide with loss of CD24. We identified and validated CD24 as a direct and functional novel target of miR-146a and found that CD24 over-expession rescued miR-146a mediated tumorigenicity. We observed a unique negative co-relation between CD24 and β-catenin under the supervision of miR-146a by positively regulating AKT activity. We show that AKT inhibition is actually conferred by CD24 that subsequently leads to β-catenin degradation. In agreement to the previous findings, we acknowledged a positive feedback loop in β-catenin mediated transactivation of miR-146a that possibly contributes to stem cell maintence. Taken together, these results provide a proof-of-principle that miR-146a plays a key role in sustaining CD44high CD24low status in OSCC.
Sangeeta Ghuwalewala is a PhD student in Indian Institute of Chemical Biology, Kolkata, under the guidance of Dr. Susanta Roychoudhury. She did her graduation in Microbiology from St. Xavierʼs Collegiate School, followed by M.Sc. Biotechnology, University of Calcutta. She has first research experience during a 2-month summer research working under Dr. Gaiti Hasan, at National centre of Biological Sciences, Bangalore. She has qualified CSIR-NET December 2009 in first attempt with an all India 23rd rank and was invited for the prestigious SPM fellowship. She works in Cancer Stem Cells and its epigenetic involvement in tumorigenesis. She has explored various aspects of tumor heterogeneity and went through quite a diverse type of research endeavours including establishment of Primary Cell line from tumors, Next Generation Sequencing, Transcriptomics as well as study of miRNA and chromatin regulatory mechanisms. She has also attended a course held in NCBS “Hands on Workshop on Epithelial Stem Cell Biology” and also have a brief exposure to NGS data analysis. She owned a “Best Poster Presentation Award” for part of her work on “PRC2 mediated CSC reprogramming” in Indian Association for Cancer Research, 2014. She has one publication in “Stem Cell Research” under revision and a few more to be communicated.