The Effects of Cisplatin on MTOR, AKT, CCND1 and STAT3 mRNA Expressions on HeLa Cells

WOS: 000505071500002Objective: Cervical cancer is one of the most tumors seen in womans and second leading cause of cancer death in women. Cisplatin (CDDP), a platin based compound, is the most important chemotherapeutic agent and effectively used for the treatment of sarcomas and solid tumors. It binds to the DNA with crosslinks and leads inhibition of replication, causing DNA damage. As a result of this action, apoptotic pathways are induced and cell death occurs. In our study, we aimed to investigate the effect of CDDP on relative mRNA expression of mTOR, AKT, Cyclin D1 (CCND1) ve STAT3 on cervical cancer cell line. Method: In this study, HeLa cells were treated with different concentrations of CDDP at 24 and 48 hours. Cell viability was determined by XTT method. Moreover, after treatment with selected doses of cisplatin, quantitative mRNA expression of mTOR, AKT, CCND1 and STAT-3 genes was analyzed using Real-Time PCR. Results: IC50 concentration of CDDP was found to be about 60 mu M for 24h and 8 mu M for 48h treatment. Moreover, all analyzed genes' expression was shown to diminish only after 24 h treatment. On the other hand, no statistically significant change was found after 48 h cisplatin exposure with respect to quantitative mRNA expression. Conclusion: In summary, different mRNA expression pattern was found after CDDP treatment regarding to exposure time. Our study has been contributed the literature in terms of detecting the effect of conventional chemotherapeutic CDDP on cell survival pathways

The Role of Matrix Metalloproteinase-2 Promoter Polymorphisms in Coronary Artery Disease and Myocardial Infarction

YILMAZ, AKIN/0000-0002-4368-0777WOS: 000288850800002PubMed: 21142815The matrix metalloproteinase (MMP) family are key enzymes involved in the breakdown of the extracellular matrix in normal physiological processes, including tissue remodeling, and disease processes, such as arthritis and metastasis. The promoter polymorphism in the MMP2 gene may be responsible for multiple diseases related to extracellular matrix degradation. Therefore, we aimed to investigate the relationship between genotypes or haplotypes of -1575 G/A, -1306 C/T, -790 T/G, and -735 C/T promoter polymorphisms and coronary artery disease (CAD) with or without myocardial infarction (MI) history. This study included 298 patients with angiographically confirmed CAD and 299 age matched controls. Genomic DNA was isolated from whole blood and genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism method. No significant associations were found between -1575 G/A, -1306 C/T, and -790 T/G polymorphisms and CAD with or without MI history. However, the frequency of the -735 TT genotype was significantly lower in the controls than in the patients with MI alone when compared with the CC genotype (p = 0.021). Only the distribution of the ACGC haplotype in CAD patients exhibited a significant difference than that in controls (p < 0.05). The distribution of other haplotypes did not differ between CAD patients and controls. The present investigation is the first report to detect an association between MMP2 promoter polymorphisms and CAD with or without MI history in the Turkish population. Further case-control studies in CAD development might be contributed to clarify the role of these polymorphisms.Gazi UniversityGazi University [11/2004-84]This study was partially supported by the Gazi University Research Fund as a project with code number 11/2004-84

Investigation of Ocular Neovascularization-Related Genes and Oxidative Stress in Diabetic Rat Eye Tissues After Resveratrol Treatment

Changes in vascular endothelial growth factor (VEGF), angiotensin-converting enzyme (ACE), matrix metalloproteinase (MMP)-9, and endothelial nitric oxide synthase (eNOS) mRNA expression profiles and oxidative stress in the eye tissue microenviroment may have important roles in ocular neovascularization and permeability in proliferative diabetic retinopathy. The present study investigated the effects of resveratrol (RSV) treatment on the mRNA expression profile of VEGF, ACE, MMP-9, and eNOS, which are associated with vascular neovascularization, and glutathione, protein carbonyl, and nitrite-nitrate levels, which are markers of oxidative stress in eyes of diabetic rats. Twenty-four Wistar albino male rats were divided into four groups. After diabetes induction with streptozotocin (10 mg/kg/day) RSV was administered to the RSV and diabetes mellitus (DM) + RSV groups for 4 weeks. The mRNA levels were measured by quantitative real-time polymerase chain reaction assay, and biochemical estimations were determined with spectrophotometric assays in eye homogenates. The mRNA expression levels of VEGF, ACE, and MMP-9 were increased in the DM group compared with the control group, and RSV treatment decreased their mRNA levels. Expression of eNOS mRNA was increased in the RSV and DM groups and decreased in the DM + RSV group. Nitrite-nitrate levels and protein carbonyl content were increased and glutathione levels were decreased in the DM group compared with controls. Consequently, these data suggest that RSV suppressed the expression of eNOS, which is actively involved in the inflammation and healing process in chronic diabetes. Although oxidative stress was increased in eye tissue from diabetic rats, mRNA levels of VEGF, MMP-9, and ACE genes associated with vascular remodeling did not change in diabetic eyes

Effects of Meloxicam, Alone and in Combination With Chemotherapeutic Agents on the Raf/Mek/Erk Pathway in Raji Cells

WOS: 000344634100027Background: Non-steroidal anti-inflammatory drugs (NSAIDs) have been reported to induce apoptosis and inhibit cell proliferation in tumor cells when combined with conventional chemotherapeutic agents. We aimed to investigate the effect of NSAIDs (meloxicam and lornoxicam) plus conventional chemotherapeutic agents on RAF/MEK/ERK pathway in Burkitt's lymphoma cell line (Raji). Methods: Raji cells were treated with meloxicam, lornoxicam, carboplatin, gemcitabine and 5-fluorourasil (5-FU) for 24h. Then, the cells were treated with selected low and high doses of meloxicam alone or in combination with two different gemcitabine concentrations. Presence of apoptotic cells were assessed using fluorescence microscopy. mRNA expression levels of RAF1, MEK1, ERK1 and ERK2 genes were analyzed with quantitative real time PCR. Results: Apoptotic and anti-proliferative effects were not observed by carboplatin, 5-FU and lornoxicam. However, meloxicam and gemcitabine treatment resulted in a decrease in viability. Therefore, we examined the effects of low and high doses of meloxicam, gemcitabine and their combinations on this pathway. The antiproliferative effects of gemcitabine were not significantly enhanced in the presence of meloxicam. Furthermore, mRNA expression levels of RAF1, MEK1 and ERK1/2 genes were downregulated in gemcitabine alone and upregulated in meloxicam alone. Interestingly, the decrease shown in low and high doses of gemcitabine alone reversed in low dose meloxicam plus gemcitabine treatment. Conversely, downregulation of these genes by high dose gemcitabine did not change with meloxicam treatment. Conclusion: We believe that it may be fundamental to further evaluate this inhibitory drug interaction

Lack of Association Between Matrix Metalloproteinase-9 and Endothelial Nitric Oxide Synthase Gene Polymorphisms and Coronary Artery Disease in Turkish Population

7th Balkan Meeting on Human Genetics -- AUG 31-SEP 02, 2006 -- MACEDONIAWOS: 000267285500006PubMed: 19435423Polymorphic variants of genes encoding proteins involved in vascular remodeling may genetically diverge among different populations and play a role in the susceptibility to the coronary artery disease (CAD). MMP-9-1562 C/T (rs3918242), eNOS T-786C (rs2070744), and Glu298Asp (rs1799983) are among the most studied of these polymorphisms. The aim of this study was to determine the relationship between CAD and these polymorphisms in the Turkish population. The analysis included 146 CAD+ and 122 CAD- individuals. Genomic DNA was isolated from whole blood and genotyping was performed by the PCR-RFLP method. No significant associations were found between -1562 C/T (p=0.557), Glu298Asp (p=0.432), and -786 T/C (p=0.055) polymorphisms and CAD. The distribution of each haplotype also did not differ between CAD+ and the CAD- samples (p>0.05). The present investigation is the first to study an association between -1562 C/T polymorphism and CAD in the Turkish population. In conclusion, no appreciable differences between CAD+ and CAD- samples were found in terms of polymorphisms mentioned above

Association between TNF-beta NcoI (A252G) gene polymorphism and type I diabetes mellitus in Turkish population

WOS: 000314288600004Objective: Diabetes mellitus is a common metabolic disorder characterized by hyperglycemia. Cytokines induce signal transduction and secondary messenger pathways after binding their specific ligands. In this way, these proteins control and also modulate activity of the target cells. Tumor necrosis factor plays an important role in diabetes and is a member of the family of cytokines. In this study, we aimed to investigate the effect of A252G (TNF-beta NcoI, rs909253) polymorphism in the tumor necrosis factor beta gene on the Type I Diabetes Mellitus in the Turkish population. Materials and Method: Distribution of A252G polymorphism was determined in 96 patients with Type I diabetes and 101 healty individuals by PCR-RFLP. Results: When obtained data was evaluated, the A allele of A252G polymorphism were observed more frequently than G allele in patients and controls. Moreover, G alleles were observed more frequently among patients than in controls. This high frequency of G allele found in patients demonstrates a 1.6 fold increased risk of susceptibility to type I diabetes in Turkish population (P=0.032). However, there was no significant difference between patients and controls in terms of the distribution of genotypes (P=0.132). Conclusion: As a result, although presence of a G allele demonstrated an increased risk of susceptibilty to type I diabetes, TNF-beta A252G polymorphism's genotype distrubution was not associated with type I diabetes in Turkish population. Polymorphisms involved in the development of Type I Diabetes Mellitus results in better understanding of the development and the progression of disease