miRNA-mediated apoptosis activation through TMEM 48 inhibition in A549 cell line.

Lung has critic function in gas exchange, supplying oxygen to all cells. Rapid metastasis and the high rate of mortality characterises lung cancer. There are two types of this disease, small cell and non-small cell, which differs from each other according to histopathologic features. To date, many therapeutic approaches have been developed to destroy this deadly type of cancer, which one of them is mRNA targeted therapies through miRNA. miRNAs are 19-25 base paired molecules be able to suppress and destruct mRNA and found to be involved in development and progression of lung cancer. Transmembrane Protein 48 (TMEM48) is localised on nuclear pore complex and plays critic roles in nuclear traffic. Known that TMEM48 gene overexpressed in non-small lung cancer cells. Growing TMEM48 suppressed therapeutic studies indicated that decreased TMEM48 level might reveal a therapeutic effect for non-small cell lung cancers. TMEM48 studies based on the same strategy of gene-silencing, however, to our knowledge, any report has been published evaluates TMEM48's regulation by miRNAs. We aimed to clarify if miR-421 might be therapeutic player for non-small cancer cell lines (A549), hereby we suppressed TMEM48 by miR-421 and performed advanced molecular tests. Consequently, we recorded that while miR-421 is significantly suppressing TMEM48 expression; it increased apoptotic and tumor suppressor players CASPASE 3, PTEN and TP53 in A549 line, which is consistent with Annexin V - PI results: 30,6% of A549 observed to be apoptotic - 68,5% of A549 was in GO/G1. Our study indicated that miR-421 can suppress TMEM48 so that leads the cells to apoptosis. But it is not entirely clear how miR-421 triggers apoptosis and whether it interacts with the other cellular death pathways in A549.

Detection of CYP1A1 and GSTP1 gene polymorphisms in bladder cancer patients in a Turkish population using a polymerase chain reaction-restriction fragment length polymorphism method.

OBJECTIVE: Understanding genetic polymorphisms might facilitate the analysis of differences between individuals in their susceptibility to developing cancers as a result of environmental carcinogens. Skin, lung, colon and bladder cancers emerge from biological defects in GSTM1, GSTT1 and GSTP1 gene expressions. In this study, we aimed to investigate whether there was an association between CYP1A1 and GSTP1 gene polymorphisms and bladder cancer in a Turkish population. MATERIAL AND METHODS: Blood samples were collected from 120 individuals (60 patients with bladder cancer and 60 healthy individuals), and their DNAs were isolated. A polymerase chain reaction-restriction fragment length polymorphism (PCR - RFLP) method was used to detect the frequencies of CYP1A1 NM_000499.3: c.*1189T > C and GSTP1 NM_000852.3: c.313A > G polymorphisms in bladder cancer patients. RESULTS: The frequency of the CYP1A1: c.*1189 TC genotype and C allele were significantly different between bladder cancer patients and healthy individuals (p=0.001 and p=0.005, respectively). However, there was no significant difference for the GSTP1: c.313 AG genotype or G allele between both study groups (p=0.699 and p=0.360, respectively). CONCLUSION: A polymorphic site of the CYP1A1 gene might be involved in the development of bladder cancer. However, the investigated GSTP1 polymorphic site did not represent an important risk factor for the development of bladder cancer in a Turkish population.

Genetic polymorphisms of hspa1b and hspa1l in infertile men.

OBJECTIVE: To investigate the relationship between the HSP 70 genepolymorphism and primary infertility in males with normal sperm-parameters. METHODS: The case-control study was conducted in Sanliurfa, Turkey, from September 2010 to August 2011and comprised infertile males as cases and healthy fertile controls. Deoxyribonucleicacid was isolated from the blood of both groups, and polymorphisms of the HSPA1B gene (NM_005346.4, GI: 3304):c.1059G>A, (PstI G>A; dbSNP: rs1061581G>A) and HSPA1L gene (NM_005527.3, GI: 3305) c.1478C>T (NcoIC>T, dbSNP: rs2227956) were analysed with polymerase chain reaction-restriction fragment length polymorphism technique. SPSS version 11.5 was used for statistical analysis. RESULTS: Of the 140males in the study, 68(28.5%) were infertile cases and 72(51.4%) fertile controls. There was no statistically significant difference between GA (heterozygous) and AA (homozygous, polymorphic) genotypes of the c.1059G>A polymorphic point of the HSPA1B gene or between the A allele of the cases and controls (p>0.05). There was no statistically significant difference between the CT (heterozygote) and TT (homozygous, polymorphic) genotypes of the c.1478C>T polymorphic area of the HSPA1Lgene or between the T alleles of the cases and the controls (p>0.05). CONCLUSIONS: Infertility in males with normal sperm parameters was not significantly associated with HSPA1B:c.1059G>A and HSPA1L:c.1478C>T gene polymorphisms. Further prospective studies with larger sample sizes and different gene groups are required to clarify the issue.

Lack of Association between PTPN22 Gene +1858 C>T Polymorphism and Susceptibility to Generalized Vitiligo in a Turkish Population.

BACKGROUND: Vitiligo is an autoimmune polygenic disorder characterized by loss of pigmentation due to melanocyte destruction. The PTPN22 gene +1858 C>T single nucleotide polymorphism (rs2476601) has been shown to be associated with various autoimmune disorders. OBJECTIVE: The aim of this study was to investigate whether the PTPN22 gene +1858 C>T single nucleotide polymorphism is associated with susceptibility to generalized vitiligo in a Turkish population. METHODS: One hundred and seven patients with generalized vitiligo, and one hundred and twelve gender-, age-, and ethnic-matched controls were enrolled in the study. Genotyping was done by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The PTPN22 +1858 C>T genotype and allele frequencies of the generalized vitiligo patients did not differ significantly from those of healthy controls. CONCLUSION: We found no association between the PTPN22 +1858 C>T gene polymorphism and vitiligo susceptibility in Turkish generalized-vitiligo patients.

Beta-globin gene mutations in children with beta-thalassemia major from Sanliurfa province, Turkey.

OBJECTIVE: The prevalence of ß-thalassemia in Sanliurfa province, Turkey is reported to be 2.6%-3.7%, whereas nation-wide the frequency of ß-thalassemia is 2%. This study aimed to identify the most frequent ß-thalassemia mutations in Sanliurfa province. METHODS: In total, 22 mutations were investigated in 115 pediatric patients with ß-thalassemia using a commercially available reverse dot blot platform. RESULTS: The study included 60 male and 55 female patients with a mean age of 7.3±4.6 years (range: 1-17 years). In total, 76% of the patients had consanguineous parents. In all, 16 different mutations were observed in the 115 patients. IVS-1-110 (G-A) (29.1%), IVS-1-1 (G-A) (13.9%), codon 39 (C>T) (10.4%), and codon 8 (-AA) (9.1%) accounted for 62.5% of all the ß-thalassemia mutations, and 6% of the patients had 2 different thalassemia mutations. According to the present results, IVS-1-110 (G>A) was the most frequent mutation observed in the patients from Sanliurfa province, as in other geographical regions of Turkey. In addition, the following 34 compound heterozygote mutant alleles were observed; IVS-1-1 (G>A)/IVS 2.848 (n=4), codon 39 (C>T)/codon 8 (-AA) (n=2), codon 6 (-A)/IVS 1.5 (G>C) (n=2), IVS-1-110 (G>A)/IVS-1-1 (G>A) (n=2), IVS-1-110 (G>A)/codon 8 (-AA) (n=1), IVS-1-110 (G>A)/codon 39 (C>T) (n=1), IVS-1-110 (G>A)/IVS-1-6 (T>C) (n=1), IVS-1-110 (G>A)/IVS-1-5 (G>C) (n=1), IVS-1-110 (G>A]/codon 8/9 (+G) (n=1), IVS-1-1 (G>A)/codon 39 (C>T) (n=1), and codon 8 (-AA)/IVS-1-5 (G>C) (n=1). The following ß-globin gene promoter mutations were not observed; -101 (C>T), -87(C>T), -30 (T>A), codon 15 (TTG>TGA), codon 27 (G>T) Knossos, and IVS-1-116 (G>C). In all, 5 of the 115 patients (4.3%) had an unidentified mutation. CONCLUSION: The present results illustrate the heterogeneity of ß-thalassemia mutations in Sanliurfa Province. The present findings may be of value for genetic counseling, and premarital and prenatal diagnosis in Sanliurfa province.

Monitoring of failure of chloroquine treatment for Plasmodium vivax using polymerase chain reaction in Sanliurfa province, Turkey.

Malaria is a complex disease that varies widely in epidemiology and clinical manifestation in the southeastern part of Turkey. In many regions of the world, chloroquine (CQ) has been the standard treatment for Plasmodium vivax. However, the resistance of the Plasmodium species to antimalarial drugs has become an increasing problem and a concern worldwide. Our target was to determine the Plasmodium species in the southeast region of Turkey and the therapeutic efficacy of CQ used in the treatment of malaria. Blood samples were collected from 180 patients infected with malaria before and after CQ treatment and were subjected to DNA isolation. The isolated DNA was amplified by a seminested multiplex polymerase chain reaction (SnM-PCR) including primers selected on Plasmodium small subunit ribosomal RNA (ssrRNA) genes for identification of the malaria species. The SnM-PCR results showed that only P. vivax exists in this province. It was also determined that there is a therapeutic failure to CQ in 9.5% of patients. These were the second report on identification of P. vivax and the third report on determination of the therapeutic failure in patients who used CQ to cure human malaria in the southeastern region of Turkey. Our results demonstrate that the SnM-PCR is a sensitive, specific, and a rapid tool for the differentiation of malaria species.

Detection of VDR gene ApaI and TaqI polymorphisms in patients with type 2 diabetes mellitus using PCR-RFLP method in a Turkish population.

Type 2 diabetes mellitus (T2DM) is by far the most common type of diabetes and is characterized by insulin resistance and altered insulin secretion. Some genes, such as the vitamin D receptor gene (VDR, NM_001017535; GI: 7421), involved in its metabolic pathway have been regarded as good candidates for T2DM. In this study, we investigated whether there was an association of VDR: g.59979G>T or c.1025-49G>T (ApaIG>T) and g.60058T>C or c.1056T>C (TaqIT>C) polymorphisms in the 3' untranslated region of VDR with T2DM in a Turkish population. We collected blood samples from 241 individuals (72 patients with T2DM and 169 healthy individuals), and their DNA was isolated. Polymorphisms of the VDR were analyzed by DNA amplification with polymerase chain reaction and endonuclease digestion with ApaI and TaqI. Body mass index was higher in T2DM patients than in control individuals. However, the frequency of g.59979TT genotype in T2DM patients was not significantly increased compared to healthy subjects (37.5% vs. 36.1%, respectively). Although the VDR g.60058CC genotype in T2DM patients (19.4%) was higher than that in healthy individuals (11.2%), there was no significant difference. In the same way, there was no difference between the groups in allele frequencies. In conclusion, our study did not provide evidence for the association of two examined VDR polymorphisms with T2DM in a Turkish population.

Identification of Leishmania parasites in clinical samples obtained from cutaneous leishmaniasis patients using PCR-RFLP technique in endemic region, Sanliurfa province, in Turkey.

Antroponotic cutaneous leishmaniasis (ACL) is an endemic disease and one of the major health problems in Sanliurfa province located in the southeastern region of Turkey. Leishmania tropica is confirmed as the causative agent of ACL in this region. In Sanliurfa city alone, the recorded total cases of ACL were 6,817 between 2001 and 2006. We aimed to determine the effectiveness of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identification and differentiation of the Leishmania parasite in comparison to direct microscopic examination of clinical samples. The lesion exudates were collected from 51 ACL suspected patients and used for smear-slide preparations and DNA isolation. The isolated DNA was amplified by PCR, including primers selected on repetitive DNA for identification of a Leishmania subgenus, and the amplified DNA was restricted by HaeIII restriction endonuclease. The PCR-RFLP results showed that only L. tropica exists in this province. It is also determined that the positivity rate with PCR was higher (96%) than by microscopic examination (64%) in the diagnosis of ACL. Our results indicate that the PCR-RFLP method is more sensitive and specific for the detection and differentiation of agents of ACL in this area.

Investigation of IVS14+ 1G > A polymorphism of DPYD gene in a group of Turkish patients with colorectal cancer.

BACKGROUND: Dihydropyrimidine dehydrogenase (DPD) is a critical enzyme in the catabolism of 5-fluorouracil (5-FU), a drug frequently used in cancer therapy. One of the possible causes of severe 5-FU toxicity is genetic polymorphisms in the DPYD gene, such as IVS14+1G > A. In this study we aimed to investigate the frequency of the IVS14+1G > A mutation in the DPYD gene in Turkish patients with colorectal cancer (CRC) and healthy controls. MATERIALS AND METHODS: Blood samples were collected from 218 individuals (56 patients with CRC and 162 healthy individuals), and the DNA was isolated. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect the frequency of the IVS14+1G > A mutation in our population. RESULTS: The IVS14+1G > A mutation (heterozygous) in the DPYD gene was identified in two healthy subjects in this Turkish population. CONCLUSION: The apparently high prevalence (allele frequency of 0.6%) of the IVS14+1G > A mutation warrants genetic screening for this mutation in cancer patients before the administration of 5-FU.