Investigation of New Benzimidazole Derivative Compounds Effects on A549 Cell Line

Abstract Chronic inflammation is a common indication of several diseases, e.g. asthma, chronic obstructive pulmonary disease (COPD), atherosclerosis, etc. Benzimidazole derivatives are preferable compounds to design new analgesic and anti-inflammatory substances due to their unique biological features. We aimed to investigate the effect of a newly synthesized benzimidazole derivative, ORT-83, on A549 human lung adenocarcinoma cell line. ORT-83 was synthesized, and a non-cytotoxic concentration of ORT-83 on A549 cells was detected with MTT assay. To analyze the anti-inflammatory effect of ORT-83, an inflammatory cell culture model was established by stimulating A549 cell line with IL1-β (10 ng/ml). After 2 hours of treatment with IL1-β to induce inflammation, A549 cells were exposed to ORT-83 (0.78 µg/ml) for 24 hours. Thereafter gene expression analyses were performed with qRT-PCR. We found that ORT-83 significantly suppressed the gene expression levels of the proinflammatory cytokines; IL-6, NFkB, and TNF-α. However, the increased levels of IL-10 (2.8 folds) by IL-1β induction did not change after ORT-83 and/or dexamethasone (Dex: positive control) treatments. While Dex; a COX-2 inhibitor, reduced the COX-2 expression level in inflammatory cells from 10.03 folds to 0.71 folds, ORT-83 reduced its level to 4.37 folds. iNOS expression levels did not change in any experimental groups. In conclusion, we showed that ORT-83 exerted its anti-inflammatory effects by repressing the gene expression of proinflammatory cytokines in the inflammation-induced A549 cell line. Although ORT-83 had a weaker COX-2 inhibitory effect compared to Dex, it was shown to be still a strong anti-inflammatory compound.

IRS-2 G1057D polymorphism in Turkish patients with colorectal cancer.

INTRODUCTION: Gene polymorphisms have a broad range of analysis, but are of particular use in molecular medicine due to their potential in revealing the genetic tendency in diseases such as cancer, heart attack etc. These studies basically depend on mutations that can be detected by proper techniques. The genes coding the insulin receptor substrate (IRS) proteins are among the most widely analysed polymorphisms in various cancer types, in which a G1057D mutation is seen. AIM: To determine the risk of colon cancer by analysing the IRS-2 gene polymorphism in Turkish patients. MATERIAL AND METHODS: A total of 161 newly diagnosed colorectal cancer patients were analysed and compared to 197 unrelated healthy controls. A polymerase chain reaction-based restriction fragment length polymorphism method was carried out. RESULTS: No differences were observed between the patient and control groups for both allele and genotype frequencies of the IRS-2 G1057D gene. CONCLUSIONS: Our results demonstrated that IRS-2 G1057D polymorphism is not associated with colorectal cancer in the Turkish population. This research is a preliminary and original study in Turkish patients with colorectal cancer. It also provides population-level genetic data on IRS-2 in the Turkish population. Further studies should be performed on larger number of patients and controls for more reliable results about the genetic tendency in colorectal cancer in Turkey. The study is a collaborative work of different universities and scientists.

Dopamine D2 receptor gene -141C Insertion/Deletion polymorphism in Turkish schizophrenic patients.

Schizophrenia is a chronic and neuropsychiatric disease that affects about 0.5-1% of the world's population. An increase in dopamine and dopamine D2 receptor (DRD2) gene products has been well described in schizophrenic patients. Several groups have studied the relationship between dopaminergic hyperactivity and cellular communications have obtained discordant results. Studies searching for the relationship between the schizophrenia and DRD2 gene have gained more interest. Our objective was to determine the relationships among schizophrenic symptoms in schizophrenia subtypes and severity of symptoms in terms of DRD2 gene -141C Insertion/Deletion [Ins/Del; I/D] polymorphism by PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) assay method. Genomic DNA was prepared from peripheral blood by using salt extraction method. After amplification of genomic DNA, PCR products were digested with BstNI restriction enzyme for the detection of DRD2 gene -141C Ins/Del polymorphism in 73 schizophrenic patients and 60 healthy control subjects. The allelic frequencies of the DRD2 gene -141C Ins/Del polymorphism in case and control groups were 79.5 and 77.5% for I allele; 20.5 and 22.5% for D allele respectively. There was no significant difference in frequencies of genotypes and alleles between the two groups. In schizophrenic and control subjects, there were no significant relationship in severity of the disease and schizophrenia types among the -141C Ins/Del genotypes and alleles.

Plasminogen activator inhibitor type-1 gene 4G/5G polymorphism in Turkish adult patients with asthma.

AIM: This study has been performed on asthmatic patients in the Turkish population to determine the frequency of 4G/5G polymorphism genotypes of plasminogen activator inhibitor type-1 gene, and with the aim of examining the role of this polymorphism in asthma development. METHODS: Genomic DNA obtained from 165 persons (98 patients with asthma and 67 healthy controls) was used in the study. DNA was multiplied with polymerase chain reaction using 4G and 5G allele-specific primers. Polymerase chain reaction products were assessed with CCD camera by being exposed to 2% agarose gel electrophoresis. Results were evaluated with chi-square test. RESULTS: No statistically significant difference between the groups with respect to genotype distribution was found (p > 0.05) in the study. The 4G allele frequency was indicated as 48% and 5G allele was as 52% in patients, whereas this was 50-50% in the control group. CONCLUSION: It has been established by this study that 4G/5G polymorphism genotypes of plasminogen activator inhibitor type-1 gene do not play a role in the development of asthma in the Turkish population.