Comparison of the effects of weakened mutagens of chemical Mitomycin C and physical UV on the genomes of lymphocytes

زمینه و هدف: بحرانی ترین تأثیر عوامل موتاژن بر ساختار ژنومی انسان، زمینه سازی برخی از ناهنجاریها و اختلالات با پیش زمینه ژنتیکی است. میتومایسین C و UV (اشعه فرابنفش) دو موتاژنی هستند که در مراکز بهداشتی و درمانی کاربرد وسیعی دارند, اما میزان تاثیرات و شدت و حدت آنها بر ژنوم سلول های طبیعی نامعلوم است. لذا در این مطالعه دو نوع موتاژن فوق، که بر اساس شاخص میتوزی تضعیف شده بودند، از نظر میزان تأثیر ناپایداری ژنومی مقایسه گردید تا اهمیت موتاژن های ضعیف فیزیکی یا شیمیایی، بیشتر مورد توجه قرار گیرد. روش مطالعه: به این منظور، تعداد 105 لنفوسیت خون محیطی جداسازی شده با فایکول، در فلاسکهای T25 حاوی ml 5 محیط کشت کامل F12 (20-15 درصد FCS) و میتوژن T لنفوسیت PHA (فیتوهماگلوتنین) و در حضور BrdU (Bromo deoxy Uridine) کشت داده شد. به صورتی که سه نمونه از فلاسک های سلولی، دارای غلظت های تضعیف شده ng/ml3، ng/ml6 وng/ml 9 از میتومایسین C بوده و دو نمونه دیگر از آنها، در فاصله cm20 از لامپ UV (شدت فرابنفشC، Lux420) به طور مجزا و در دو زمان 3و 5 دقیقه، تحت پرتوتابی قرارگرفته، به همراه همان تعداد سلول لنفوسیتی به عنوان شاهد، در دمای ˚C 37 انکوبه گردید. با برداشت سلول های متافازی بعد از 72 ساعت از زمان کشت و رنگ آمیزی آنها با روش SCE Sister Chromatid Exchange))، میانگین درصد تعداد تعویض های کروماتید خواهری در کروموزوم های پلاک های متافازی حاصل، مورد ارزیابی قرار گرفت. نتایج: مطالعه سلول های تیمار شده با هر دو موتاژن تضعیف شده و نمونه شاهد از نظر SCE، نشان داد که درصد تبادلات کروماتید خواهری در سلول های تیمار شده با MMC (Mitomycin C) با غلظت های ng/ml3، ng/ml6 و ng/ml9 به ترتیب43/5، 1/7 و 13/8 درصد بوده و در سلول های در معرضUV ، به مدت 3 و 5 دقیقه به ترتیب 34/4 و 8/6 درصد می باشد، در حالی که این میزان در سلول های شاهد 35/3 درصد تعیین گردید. آنالیز آماری نتایج حاصل، معنی دار بودن تمامی داده ها را نشان داده است (001/0

The Investigation of WRAP53 rs2287499 Association with Thyroid Cancer Risk and Prognosis among the Azeri Population in Northwest Iran

Background: TP53 and the oncogene WRAP53 are adjoining genes, producing p53-WRAP53α sense-antisense RNA couples. WRAP53α is indispensable for p53 mRNA regulation and p53 induction following DNA damage. Up-regulated WRAP53β can induce neoplastic transformation and cancer cell survival. All these, along with the associations of WRAP53 single nucleotide polymorphisms with tumor incidence and prognosis, highlighted an impact in human cancers. Considering the importance of WRAP53 in modulating p53, and the frequent occurrence of thyroid cancer, we examined the association of a WRAP53 SNP (rs2287499) with thyroid cancer risk and prognosis among Iranian-Azeri population. Methods: This research was done in Tabriz-IRAN in 2014. DNA samples obtained from 106 patients and 196 controls were subjected to polymerase chain-reaction-based single-strand conformational polymorphism (PCR-SSCP) analysis. Genotypes were characterized by sequencing. Correlations of desired SNP with thyroid cancer as well as age, gender, involved thyroid lobe, lymph node metastasis, tumor type, stage, and size were estimated using Chi-square (χ2) or Fisher's exact tests with a P-value less than 0.05 as significant. Results: rs2287499 is not associated with thyroid cancer predisposition. Except for gender, none of the clinicopathologic factors were significantly linked to the examined genotypes. Conclusions: rs2287499 is not a genetic risk factor for thyroid cancer. Although rs2287499 is not assessable as a biomarker to predict prognosis based on clinicopathologic parameters, the considerable association with gender suggests that this SNP may indirectly be relevant to gender-associated disease manifestation. Further investigations on distinct types of thyroid tumors are needed to fully characterize the rs2287499 status in thyroid malignancies

A novel mutation (4040-4045 nt. delA) in exon 14 of the factor VIII gene causing severe hemophilia A

Hemophilia A is an X-linked congenital bleeding disorder caused by Factor VIII deficiency. Different mutations including point mutations, deletions, insertions and inversions have been reported in the FVIII gene, which cause hemophilia A. In the current study, with the use of conformational sensitive gel electrophoresis (CSGE) analysis, we report a novel 1-nt deletion in the A6 sequence at codons 1328-1330 (4040-4045 nt delA) occurring in exon 14 of the FVIII gene in a seven-year-old Iranian boy with severe hemophilia A. This mutation that causes frameshift and premature stop-codon at 1331 has not previously been reported in the F8 Hemophilia A Mutation, Structure, Test and Resource Site (HAMSTeRS) database

Cytotoxic and apoptosis inducing effect of some pyrano [3, 2-c] pyridine derivatives against MCF-7 breast cancer cells

Anti-cancer activities of some pyrano-pyridines have been previously reported. Herein, we investigated anti-proliferative and apoptotic effects of the novel pyrano [3, 2-c] pyridine (P.P, TPM.P, 4-CP.P and 3-NP.P) compounds against MCF-7 breast cancer cells. The MCF-7 cells were cultured in the presence of various concentrations (20-200 μM) of the tested compounds for 3 days and the cell viability was determined by MTT assay. Induction of apoptosis was qualitatively assayed by acridine orange/ethidium bromide (AO/EtBr) staining, DNA fragmentation assay, as well as quantitatively by Annexin V/PI double staining and cell cycle analysis. These compounds inhibited growth and proliferation of the MCF-7 cells in a dose- and time-dependent manner. The IC50 values of P.P, TPM.P, 4-CP.P and 3-NP.P after 24 h of exposure were calculated to be 100±5.0, 180±6.0, 60±4.0 and 140±5.0 μM, respectively. 4-CP.P was determined as the most potent compound and was chosen for further studies. The result of flow cytometric cell cycle analysis indicated an increase in sub-G1 population after 72 h treatment of the cells. Furthermore, this was accompanied by exposure of phosphatidylserine (PS) in the outer cell membrane after time course of treatment with the 4-CP.P. Based on these observations, the pyrano [3, 2-c] pyridines can be regarded as a valuable candidate for further pharmaceutical evaluations

Importance of mir-411-5p in colorectal cancer

The abnormal expression of microRNAs (miRNAs) plays a key role in colorectal cancer (CRC). The present study attempted to identify the potential miRNA biomarkers of CRC due to the important role of microRNAs within the DLK1-DIO3 genomic region, especially the role of mir-411-5p in other cancers. This prospective study characterized the contribution of mir-411-5p to CRC tumorigenesis. The Real-time quantitative reverse-transcription –polymerase chain reaction was used to examine miR-411-5p expression levels prospectively in 40 pairs of samples of CRC tissues and adjacent noncancerous tissues (>2 cm from cancer tissue). Also, the relationship between miR-411-5p expression levels and clinicopathological characteristics was explored. The capability of miR-411-5p to function as a tumor marker in CRC was also examined. MiR-411-5p was significantly downregulated in a group of CRC samples compared with matched noncancerous tissues. A receiver operating characteristic (ROC) curve also showed ROC area of 68% for miR-411-5p (P value=0.006) with 70% and 65% sensitivity and specificity, respectively. According to the survey results, miR-411-5p might be considered as a tumor marker in CRC and it might be a promising therapeutic option which may help prevent CRC

Haplotype and linkage disequilibrium of TP53-WRAP53 locus in Iranian-Azeri women with breast cancer.

Among the cancer susceptibility genes, TP53 is one of the crucial genes involved in cell cycle regulations and, therefore, it greatly affects breast cancer initiation and progression. In addition, WRAP53-a natural antisense transcript-regulates TP53 transcription and, as a protein, modulates the normal cell cycle, which results in breast cancer susceptibility. In this study, we aimed to analyze a haplotype comprising four SNPs, including rs1042522, rs17878362, rs2287499, and rs2287498, which are located at 5' regions of the TP53 and WRAP53 genes, in 118 patients and 110 healthy controls of the Iranian-Azeri population. In silico studies were conducted using the SIFT, Polyphen2, Fanthmm, RNAsnp, and SNP&GO online servers. Linkage disequilibrium (LD) and D' for each combination of the markers were calculated via the Haploview program. Our results showed that the GA1CC haplotype was the most frequent in the studied population. Additionally, no significant LD between any pairwise haplotypes was observed. The GA1CC and CA2GC haplotypes were significantly associated with breast cancer susceptibility. Moreover, the in silico analysis revealed the negative effects of rs2287499 and rs1042522 on WRAP53 and P53, respectively. In conclusion, the CA1GC haplotype was strongly identified as a breast cancer risk factor, and the GA1CC haplotype was assumed to be a protective factor against breast cancer risk. Hence, these markers may potentially be used as molecular prognostic and predictive biomarkers for breast cancer

Investigation on the Binding Mode of 3, 4-Dihydropyrano[c]Chromene Derivative with Double Strand DNA

Purpose: The study on the interaction between small molecules and DNA has been very useful for investigating the structure and physical properties of DNA, elucidating the damage mechanism of DNA and significant in the design of new drugs targeted to DNA. This article describes an interaction of native calf thymus DNA (ctDNA) with a new 3, 4-dihydropyrano[c]chromene derivative, 2-amino-4-(4- chlorophenyl)-5-oxo-4H, 5H-pyrano-[3, 2-c] chromene-3-carbonitrile (4-CC) by using spectroscopic and viscometric techniques. Methods: The interaction between 4-CC and ctDNA is realized from the UV absorption spectrophotometry, viscosimetry, circular dichroism and fluorescence spectroscopic techniques which shows that the successive interaction of 4-CC with ctDNA occurs. Results: The experimental results revealed that 4-CC can interact with DNA through non- intercalative mode and the intrinsic binding constant (Kb) for 4-CC with DNA was estimated to 2.37 (±0.001) ×103 M-1. Methylene blue (MB) displacement studies revealed that 4-CC did not have any effect on MB bound DNA which is indicative of groove binding mode. Furthermore, 4-CC induces detectable changes in the CD spectrum of ctDNA as well as changes in its viscosity study corroborate the above experimental results. Conclusion: These results further advance our knowledge on the molecular aspects on the interaction of 4-CC to nucleic acids

Investigating Quantitative analysis of the gene expression of calcium/calmodulin-dependent protein kinase IV by the effect of Olibanum alcoholic extract in PC12 cell line

Background & Objective: Long-term memory depends on protein synthesis. The product of Camkiv gene promotes memory via activating its proteins. The treatment of laboratory animals by Olibanum leads to memory improvement and the recovery of Alzheimer. Therefore, the aim of this study is the evaluation of Olibanum ethanolic extract on the Camkiv expression in PC12 cells. Materials & Methods: Olibanum toxicity on the cell viability was investigated by MTT test. Cells were treated with concentrations 10,25,40,55,70 and 85 μg/ml of extract in time intervals 12,24,48 and 72 hours and their absorption rate was measured. Then, cells were treated by concentrations 2 and 20 μg/ml of extract in mentioned times. Extracted RNA was converted into cDNA and real-time PCR performed. Results: Cell death was raised by increasing time and concentration of extract treatment. IC50 values were obtained as 71.01, 52.95, 21.05 and 13.85 μg/ml in 12, 24, 48 and 72 hours of treatment, respectively. Besides, concentrations 2 and 20 μg/ml significantly increased Camkiv expression following 24 hour treatment. The maximum expression of Camkiv was observed in 48 hour treatment. The effect of Olibanum on gene upregulation was stable until 72 hours. Conclusion: The Olibanum ethanolic extract can remarkably upregulate Camkiv expression for a long time. These results are consistent with the previous studies indicating the effect of Olibanum on upregulation of Bdnf, Camkiv‌-downstream gene. However, regarding the existence of the two-direction pathway in the expression regulation of Bdnf and Camkiv‌, comprehensive studies are required to determine exact mechanism of Olibanum function in the brain

An Association Analysis of Reelin Gene (RELN) Exon 22 (G/C), Rs.362691, Polymorphism with Autism Spectrum Disorder among Iranian-Azeri Population

Background Autism spectrum disorder (ASD) is a intricate childhood neuropsychiatric disorder that is described by deficits in communication of verbal and non-verbal, reciprocal social interactions, stereotypic behaviors, interests, and activities. The studies of post-mortem neuro-anatomical anomalies have indicated that migration alterations could occur early during development (first trimester) in autistic brain. Since the Reelin gene, plays a crucial role in these migratory processes, it is subsequently considered as a potential candidate gene for autism. Materials and Methods In this case-control study, we recruited 74 patients with ASD and 88 healthy controls from Iranian-Azeri Population. Genomic DNA isolated from blood leukocytes of cases and control individuals by the proteinase K and using salt-out method. Single nucleotide polymorphisms (SNP) genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Results The allele and genotype frequencies did not show significant difference between autistic and control groups (P>0.05). No significant relationship was observed between the genders and genotypes in autism group (P>0.05). Conclusion The current study showed that the SNPs rs362691 could not be used as a useful molecular biomarker to predict genetic susceptibility for ASD among Iranian-Azeri patients