Antiproliferative and anti-inflammatory activities of a dichloromethane-methanol extract from Pterocarpus mildbraedii leaves

EACR24 Poster Sessions / European Journal of Cancer 61, Suppl. 1 (2016) S9–S218 in CRC cells. DDR1 is a poorly-characterized receptor TK, whose ligands are collagens, ones of the major components of the extracellular matrix. Consistent with this hypothesis, our data demonstrate that DDR1 is abundantly expressed in CRC and promotes invasive and metastatic properties of CRC cells. Importantly, this pro-invasive function of DDR1 absolutely requires receptor TK activity. We next demonstrated that nilotinib responses observed in CRC are largely mediated through inhibition of DDR1 TK activity: remarkably, expression of the nilotinib-resistant DDR1/T807I receptor, in which the gatekeeper threonine in the nilotinib-binding pocket was mutated into the bulkier isoleucine, counteracts all the inhibitory effects of this drug in CRC cells. We also found a dramatic anti-metastatic activity of nilotinib in mice that have already developed DDR1-dependent metastatic nodules, thus revealing an unsuspected role of DDR1 during late metastasis. The clinical relevance of these observations was supported by transcriptomic data demonstrating that the level of DDR1 expression is associated with a shorter relapse free survival in patients with CRC and by biochemical analyses showing that DDR1 catalytic activity is highly increased in metastatic nodules compared to the primary tumor and the healthy tissue of the same patient. Additionally, phosphoproteomic analyses revealed the RAS independent nature of DDR1 signaling and pointed BCR as an essential DDR1 substrate for its pro-invasive activity. Altogether, these data suggest that the targeting of DDR1 by nilotinib may be of therapeutic value in metastatic CRC, including those harboring a RAS oncogenic signaling. No conflict of interest. 663 Antiproliferative and anti-inflammatory activities of a dichloromethane-methanol extract from Pterocarpus mildbraedii leaves C. Otuechere1 , G. Santosh2 , O. Farombi3 . 1 Redeemer’s University, Department of Chemical Sciences- Biochemistry Programme, Ede TownOsun State, Nigeria, 2 Indian Institute of Integrative Medicine, Cell Signaling and Pharmacology, Jammu, India, 3 University of Ibadan, Department of Biochemistry, Ibadan- Oyo State, Nigeria Background: Inflammation is related to several chronic diseases, including cancer whose incidence is progressing rapidly in sub-Saharan Africa. Pterocarpus mildbraedii is a special leafy vegetable in Nigeria. Our aim was to evaluate the anticancer and anti-inflammatory properties of the dichloromethane-methanol extract from Pterocarpus mildbraedii (PME) in vitro and in vivo experimental models respectively. Materials and Methods: The antiproliferative activity was evaluated on 5 human cancer cell lines namely, promyelocytic leukemia (HL-60), monocytic leukemia (THP-1), prostrate adenocarcinoma (PC-3), colon adecarcinoma (COLO-205) and lung carcinoma (A549) by 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide (MTT) assay. Furthermore, we evaluated the anti-inflammatory activity of the extract against pesticide-induced alteration of inflammatory proteins in albino rats. Results: PME had the most anti-growth activity on the cancer cell line HL-60 cells with an IC50 value of 8.5 mg/ml. The extract also showed much less cytotoxicity against the other cell lines. The increased expressions of COX-2 and iNOS, accompanied by NFúB up-regulation, and correlating with elevated levels of myeloperoxidase and nitric oxide in the liver of pesticide-treated animals were ameliorated by PME at a dose of 200 mg/kg. Total phenolic and flavonoid contents were calculated using the Folin-Ciocalteu and the aluminum chloride colorimetric methods and found to be 0.215±0.005 and 0.082±0.003 mg GAE/g respectively. PME showed a maximum 1,1-diphenyl2-picryl hydrazyl (DPPH) radical scavenging activity of 55%. Conclusions: The results of these assays indicated that Pterocarpus mildbraedii can be a candidate for prevention and treatment of many diseases related to inflammation and malignancy. No conflict of interest.

Sunday 10 July 2016 Poster Session

Molecular and Genetic Epidemiology I 666 Cancer of unknown primary is associated with diabetes ¨ Sweden X. Li1 . 1 Lund University, Clinical Research Center, Malmo, Background: Both type 1 diabetes (T1D) and type 2 diabetes (T2D) increase in incidence worldwide. T2D is associated with many cancers. However, no data are available on cancer of unknown primary (CUP), a relatively common, fatal cancer for which tobacco smoking is the only known risk factor. At diagnosis of CUP metastases are found in various organs which has implications for prognosis. We carried out a nationwide study on the association of CUP with T1D and T2D. Patients and Methods: 32,600 T1D patients and 178,000 T2D patients were identified from the national Hospital Discharge Register, Outpatient Register and Primary Health Care Register and these were linked to the Swedish

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Cancer Registry. Standardized incidence ratios (SIRs) were calculated for CUP from 1997 through 2010 using anyone without diabetes as a reference. Results: The SIR of CUP in 421 diabetic patients was 1.71, highest for CUP with liver (2.17) and respiratory system metastases (1.95). The SIR was 2.91 for T1D but with small number of patients, 1.38 for T2D with insulin treatment and 1.78 for the main group of T2D. CUP with liver and respiratory system metastases were increase for each diabetic type but for T2D also CUP with gastrointestinal and bone metastases were increased. The highest SIRs for CUP were recorded in the three types of diabetes during the first year of follow-up, most likely due to diagnostic bias during medical work-up. Conclusions: The results provide the first demonstration that CUP is one of the cancers associated with diabetes, with definite evidence on T2D. CUP has a poor prognosis, which may be even worse when diabetes is the underlying co-morbidity. A mechanistic question for future work is to resolve whether diabetes promotes primaries that escape detection or their metastatic spread. No conflict of interest. 669 Effects of WW domain-containing oxidoreductase (WWOX) gene polymorphisms in the Kozak sequence on the risk and progression of oral cancer C. Ying-Erh1,2 , S.F. Yang2 , C.W. Lin3 . 1 Chung Shan Medical University, School of Medicine-, Taichung City, Taiwan, 2 Chung Shan Medical University Hospital, Department of Medical Research, Taichung, Taiwan, 3 Chung Shan Medical University, Institute of Oral Sciences, Taichung, Taiwan Introduction: In Taiwan, oral cancer is the fourth leading cancer in males and is associated with exposure to environmental carcinogens. WW domaincontaining oxidoreductase (WWOX), a tumor suppressor gene, is associated with the development of various cancers. We hypothesized that genetic variants of WWOX influence the susceptibility to oral cancer. Material and Method: We genotyped 5 single nucleotide polymorphisms of WWOX from 761 male patients with oral cancer and 1199 male cancer-free controls. The association between WWOX expression and WWOX genetic polymorphism was analysed in the Encyclopedia of DNA Elements (ENCODE) data from the NCBI gene database and confirmed by quantitative real-time PCR. Results: We observed that the WWOX rs11545028 polymorphism carriers involving the TT or CT + TT genotype were associated with oral cancer susceptibility. Furthermore, patients with advanced-stage oral cancer were associated with a higher prevalence of WWOX rs11545028 polymorphisms with the variant genotype TT than did patients with the wild-type gene. An additional integrated in silico analysis confirmed that rs11545028 affects WWOX expression, which significantly correlates with tumor expression and subsequently with tumor development and aggressiveness. Conclusion: In conclusion, genetic variants of WWOX contribute to the occurrence of oral cancer, and the findings regarding these biomarkers provided a prediction model for risk assessment. No conflict of interest. 670 The effect of the CYP1A1*2A variant on colorectal cancer susceptibility in a British population E. Ozta¸s1 , G. Ozhan1 , A.K. Daly2 . 1 Istanbul University, Faculty of PharmacyDepartment of Pharmaceutical Toxicology, Istanbul, Turkey, 2 Newcastle University, Institute of Cellular Medicine, Newcastle upon Tyne, United Kingdom Background: Colorectal cancer (CRC) is keeping its majority on public health for decades due to increasing incidence and high mortality despite of the overall improvements in diagnostics and treatment. Indeed, it is well known CRC has a complex aetiology and been still not well defined. Genetic polymorphisms on the biotransformation or DNA repair genes alone and as a contributor to dietary and environmental factors probably affect carcinogenic process. CYP1A1 gene is associated with tobacco-related cancers, including CRC, and extensively studied due to their key role on to metabolic activation of polycyclic aromatic hydrocarbons (PAHs) such as benzo[a]pyrene. CYP1A1*2A (6235 T/C, rs4646903, MspI) variant thought to be associated with increased risk of CRC, however, results of previous studies are conflicted [1,2]. In the present study, we examined a possible association between the CYP1A1*2A allele and CRC and the effect of cigarette smoking on this risk in a British population. Material and Methods: A prospective case–control study with British participants from Newcastle and North Tyneside Hospitals. 200 cases diagnosed with histologically confirmed adenocarcinoma of colon or rectum and 254 age-sex matched controls were included [3]. Genotyping of CYP1A1*2A was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Genotyping analyse was performed based on a dominant model with the wild type was chosen as reference group. Data comparisons were done by using Fisher’s exact test. Results: Genotyping analysis based on dominant model showed that individuals with C allele had about 1.5-fold risk of developing CRC