Tehran University Medical Journal

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Background: Gastric cancer is the most prevalent cancer with poor survival in gastrointestinal tract. Caspase 3 and 9 play an important role in the development and progression of cancer. Polymorphisms in the genes for these enzymes can affect gene activity and thus may influence susceptibility to gastric cancer. In this study, caspase 3 and 9 genes polymorphisms in patients with gastric cancer were examined.
Methods: In a case - control study, 100 patients with gastric cancer and 100 healthy individuals were evaluated in the region rs4647601: G> T for caspase-3 and -1263 A> G gene promoter for caspase 9. DNA extraction was performed from whole blood according to manufacture protocol. RFLP-PCR method was carrying out for detection of caspase 3 and 9 genes genotype in two groups.
Results: In this study, 143 men and 57 women were evaluated. All of them were selected from the same race and geographical area. The results indicated an increase of the mutant G allele in the control group, which leads to a decreasing in the incidence of gastric cancer (P<0.0001, OR: 0.096, (%0.95CL) =0.04-0.23).
Conclusion: It seems that screening of -1263 A> caspase 9 polymorphism could be a useful marker in personal sensitivity to gastric cancer and help to cancer treatment and prevention process. It is concluded that caspase gene variation may be a diagnostic factor in the gastric cancer.

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Background: Helicobacter pylori has been classified as the class I carcinogenic agent by world health organization. Colonization of the human stomach with H. pylori is a risk factor for gastroduodenal diseases. The secreted vacA toxin is an important H. pylori virulence factor that causes multiple alterations in gastric epithelial cells and T cells. Several families of vacA alleles have been described, and H. pylori strains containing certain vacA types (s1 and m1) are associated with an increased risk of gastric disease, compared to strains containing other vacA types (s2 and m2). We examined the association between H. pylori vacA s alleles and gastroduodenal diseases in Iran. Methods: A total of 149 H. pylori strains were obtained from patients with gastritis, peptic ulcer, and gastric cancer referring to endoscopy units of several cities in Iran. Biopsy culture and DNA extraction were performed and the frequency of vacA s alleles was investigated by using PCR amplification. Linear regression and binary logistic regression models were used to analyze the association between vacA (vacuolating cytotoxin A) s alleles and gastroduodenal diseases. Results: There was no significant association between the frequency of vacA s alleles and gastroduodenal diseases (gastritis or peptic ulcer disease and gastric adenocarcinoma (P> 0.05)). Conclusion: It is proposed that the H. pylori vacA s1 genotype could not be considered as an important determinant of gastroduodenal diseases in Iranian population and probably if s1 allele is associated with other virulence alleles of this gene, it will cause diseases.

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Background: Colorectal Cancer (CRC) is the third common cancer in the world. One of the pathways in colorectal tumor genesis is Microsatellite Instability (MSI+). MSI is detected in about 15% of all colorectal cancers. Colorectal tumors with MSI have dis-tinctive features compared with Microsatellite Stable (MSS) tumors. Due to the high percentage of MSI+ in patients with CRC in Iran, screening of this type of CRC is im-perative. In current study, two markers (BAT-26 and BAT-25) were used to determine an appropriate screening technique with high sensitivity and specificity to diagnose MSI status in patients with CRC. Methods: Allelic variation in two markers (BAT-26 and BAT-25) was analyzed in tis-sues and sera of 44 normal volunteers and tumor and matched normal mucosal tissues as well as sera of 44 patients with sporadic colorectal cancer by Real Time PCR (Hy-bridization probe) and High-Performance Liquid Chromatography (HPLC) techniques. The sensitivity and specificity of Real Time PCR and HPLC compared with sequencing as gold standard. The data were statistically analyzed using Student’s t-test and 2 or fisher exact test, where applicable with (P<0.05). Receiver-operating-characteristic (ROC) curves were used to evaluate the sensitivity and specificity. Results: The sensitivity and specificity of BAT-26 with Real Time PCR method (Hy-bridization probe) were 100% in comparison with gold standard method. Whereas the sensitivity and specificity of BAT-26 and BAT-25 with HPLC were 83%, 100% and 50%, 97%, respectively. Neither HPLC nor Real time PCR could detect circulating DNA with MSI property in sera. Conclusion: The sensitivity and specificity of real time PCR in MSI detection is the same as sequencing method and more than HPLC. BAT-26 marker is more sensitive than BAT-25 and MSI detection with Real time PCR could be considered as an accu-rate method to diagnose MSI in CRC tissues not sera.

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Background: In advanced stages, Colorectal cancer remains often refractory to classic therapies. In consequence, search for new therapeutic modalities with minimal toxicity is of particular interest in colon cancer management. In this regard, powerful growth-inhibitory effect has been shown for fish-oil derived Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) against cancer cells. In the present study, we evaluated the anti-cancer effect of EPA and DHA (n3-polyunsaturated fatty acids, n3-PUFAs) on the human colorectal cancer cell line (LS174T) on a dose-response and time-course ba-sis. Methods: LS174T cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37 ºC in a humidified incubator. Cancer cells were treated to vari-ous concentrations of EPA and DHA (50, 100, 150 µM/L) and incubated for 24-72 hours. Following treatments, dose-response and time-course cytotoxicity using viability and MTT assays were performed. Results: Viability analysis showed that 150 µM/L PUFAs decreased significantly the proliferation of treated cells, as compared to untreated cells. In this regard, cell viabil-ities were found to be %31±%5.1 and %30±%2.6 for DHA and EPA respectively. Moreover, treatment of cells with increasing concentrations of EPA and DHA signifi-cantly decreased growth rates in a dose-and time-dependent manner. Following 72 hours treatments with 150 µM/L PUFAs, growth rates were found to be %19±%5.5 and %20±%5 for DHA and EPA relative to untreated cells respectively. Conclusion: The results of this study indicate that n3-PUFAs decrease cell proliferation and could provide new approaches in malignant tumor therapeutic strategies.

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Background: Cancer is the third cause of death in Iran. Approximately 5-10% of all human cancers are thought to be caused by occupational exposure to carcinogens. Dif-fuse mesotheliomas of the peritoneum and pleura are considered "signal tumors," or pa-thognomonic of exposure to asbestos. There is also evidence that nonasbestos agents can induce malignant mesotheliomas. So, we need to study the history of exposure to asbestos in patients suffering from mesothelima to determine relationship between them in Iran, especially in occupational settings. Methods: We received the data of 272 mesothelioma's patients having been registered in 1385 to 1389 in Cancer Bureau, in Ministry of Health. One hundred fifty four of them had telephone number. Finally we could interview with 60 families and gathered data of patient's history of occupation, cigarette smoking and residence place interred in the designed questionnaire. Then we brought out the variants of age, sex, the province of birth place and the involved organs from the data of Cancer Bureau. Analysis of frequency of variants was carried out and compared once in 272 patients being registered in Cancer Bureau and then in the interviewed 60 patients. Results: Distribution of age, sex, the province of birth and living place and the involved organ were similar in cancer registry's data and the interviewed patients. Distribution of cigarette smoking and exposure to asbestos in the interviewed patients were 28% and 63% respectively. Occupational distribution of interviewed patients consisted of con-struction workers 20%, oil company employees 13.3%, administrative staff 8.3% and drivers 6.7%. Conclusion: The results of our study specified the jobs with high risks of exposure to asbestos and approved the relationship between the exposure to asbestos and the inci-dence of mesothelioma in Iran, according to researches in other countries. So the con-sumption of asbestos in Iran, like 20 other countries in the world is necessary to be banned.

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Background: Radiotherapy has been used to treat many types of cancers over the past years. Radiotherapy generates side effects on normal tissues. Radiosensitizer products provide decrease in tumor proliferation and reduce radiation dose in radiotherapy. Docosahexaenoic Acid (DHA) as an omega-3 polyunsaturated fatty acid has anti-proliferative effects on malignant cells. In this study, the effects of DHA accompanied by ionizing radiation on growth rate and survival fraction of HT29 colorectal cancer cells were evaluated. Methods: The present study was performed at the Institute of Biotechnology, affiliated to Urmia University, Urmia, Iran in the year 2013. In this laboratory experiment, ma-lignant cells were cultured in RPMI-1640 supplemented with 10% fetal bovine serum. HT-29 cells were cultured at 5105 cells/well into 6-well culture plates for overnight. Thereafter, the cells were pretreated with either 50 or 100 µM DHA for 4 hours and malignant cells were irradiated with either dose of 2 or 10 Gy. Cell viability was evalu-ated by trypan blue staining after 48 hours. Moreover, malignant cells were pretreated with either 50 or 100 µM DHA for 48 hours and irradiated with dose of 2 to 10 Gy. Thereafter, survival rate was evaluated by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay after 6 days. Results: Cell viabilities were found to be 59.8% and 17.5% for 50 µM DHA in combi-nation with doses of 2 and 10 Gy respectively. Using 100 µM DHA diminished cell vi-ability up to 47% and 13.9% following doses of 2 and 10 Gy respectively. Treatment of cells with DHA accompanied by increasing doses of γ-rays significantly diminished survival rate. In treated cells with 50 and 100 µM DHA, survival rate were measured to be 79.1%, 57.6%, 42.8%, 40.5%, 34% and 55.8%, 43.7%, 33.6%, 27.9%, 23.5% for doses of 2, 4, 6, 8 and 10 Gy respectively. Conclusion: Our study indicates that DHA decreases colorectal cancer cells prolifera-tion and could provide a new radiosensitizer drug to enhance the efficacy of colorectal cancer radiotherapy.

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Background: The aim of this study was to assess the role of consolidative intraperito-neal chemotherapy with carboplatin in decreasing relapse and increasing survival in advanced epithelial ovarian cancers, as well as evaluation of its toxicity. Methods: In this clinical trial 30 patients with epithelial ovarian cancer in stages II-IV who had complete surgery (optimal debulking surgery) received six standard cycles of intravenous carboplatin and paclitaxel. They were enrolled through non-random se-quential selection. The control patients were similar to case group in stage (II-IV) and pathology (epithelial ovarian cancer). The control group was evaluated retrospectively through hospital files. This clinical trial performed in Gynecology Oncology department in Tehran Valiasr University Hospital, during 2005-2010. They including 18 cases as the intervention group receiving intraperitoneal chemotherapy and 12 patients as the control group with only retrospective follow-up. The cases received 3 cycles of 400 mg/m2 intraperitoneal carboplatin every 21 days following intravenous chemotherapy. Relapse of disease was diagnosed as increasing or even doubling CA125 serum titer during one month, or any CA125 above 100 IU, or an abdominal or pelvic mass in ul-trasound or physical exam. Mean survival of two and five years, progression-free inter-val (PFI), overall survival (OS), relapse, demographic parameters, drug toxicities, path-ologic types of cancers in two groups were coded and compared using SPSS 14. Any P<0.05 was considered as a significant difference. Results: The mean ages of cases and controls were 52.4±8.6 and 55.1±11.5 years. The mean duration of relapse-free survival was 13±8.6 months for the cases and 9.5±4.3 months for the control patients (not statistically different, P>0.05). The mean overall survival for cases and controls were 39±16.5 and 30.8±16.2 months, respectively (no significant difference, P>0.05). The frequency of drug toxicities in the cases was 5.6%, and consisted of mild-to-moderate abdominal pain, nausea and vomiting. Conclusion: It seems that consolidation therapy with intraperitoneal carboplatin may not increase overall survival, reduce relapse rate or decrease mortality, though it does not induce considerable side effects. Since the mean survival in the intervention group was nine months more than controls, this difference may be clinically significant.

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Background: Colorectal cancer is the third most common cancer in the world. Non-coding RNA especially miRNAs have important regulatory roles in cancer. miRNAs are small non coding RNA 21-23 nucleotides long which have different levels of expression between tumors and normal tissues. This study was designed to compare expression level of miRNA-21 between Iranian population colorectal cancer tissues and normal tissue. Methods: This case-control study has performed in medical genetics department of Tehran University of Medical Sciences from January to November 2013. We used 35 samples. The samples were isolated from tumor and adjacent normal tissues of colon. Thirty-five samples were divided into different groups according to cliniopathologic features including tumor size (>4 and <4 cm), metastasis (+ and -) and stage. After small RNA extraction from tissues by small RNA purification kit the quality and quan-tity of extracted RNA was determined using spectrophotometry. cDNAs were synthe-sized and real-time polymerase chain reaction carried out. Finally expression levels were statistically analyzed by LinRegPCR and REST software. Results: miRNA-21 expression ratio in stages I, II and III were 1/804 and 4/574, re-spectively, the increase from stage III was statistically significant (P= 0.037). The ex-pression were also studied according to different clinicopathologic status of colon can-cer, tumor size (>4 and <4 cm) and metastatic (+ and -), miRNA-21 over expressed in both groups, however the increase was not statistically significant. Conclusion: In this study, we found miR-21 over-expression in advanced stage in tu-moral tissue comparing with normal adjacent tissue. This means perhaps in the future it would be possible to use miRNA-21 as an informative prognostic biomarker to guide for better treatment strategies for colorectal cancer patients. Our findings also indicate that miRNA-21 is a promising new molecular target for designing novel therapeutic strategies to control colorectal cancer.

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Background: Squamous cell carcinoma (SCC) is the most frequent oral cancer. Protec-tive effects of the consumption of vegetables and fruits on various forms of cancer in-cluding oral cancer have been determined. Tomato (Solanum lycopersicum L.) because of its lycopene and bioflavonoids contents possesses anti-carcinogenic properties. The aim of this study was to evaluate the preventive effects of tomato pulp on pre-neoplastic changes induced by 4-Nitroquinoline-1-oxid (4-NQO) in epithelial cells of lingual mucosa in the rats. Methods: Forty-eight male Wistar rats were randomly allocated into four equal groups. Group 1 served as control. Groups 2 to 4 assigned to receive 30 ppm 4-NQO in drinking water for 12 consecutive weeks. When the feeding of 4-NQO was started to the rats of groups 3 and 4, they received tomato pulp (20 and 40 ml/kg bw) daily through the oral gavage. Finally, histological evaluations for carcinogenesis were performed for tongues epithelial tissue. Results: There were no pathological alterations in epithelial tissue of lingual mucosa in control rats. In the epithelial cells of lingual mucosa of 4-NQO treated rats, premalig-nant alterations appeared after 12 weeks of the last application of the drug. Administration of tomato pulp at both doses (20 and 40 ml/kg bw) during the experiment reduced the severity of the lesions, as well as caused a significant reduction in the frequency of pre-neoplastic lesions of tongue epithelial cells (P= 0.024 and P= 0.008). The incidence of severe epithelial cells dysplasia of lingual mucosa in the high dose treatment group was significantly smaller than of low dose treatment group (P= 0.037). Conclusion: The results obtained showed that tomato pulp is effective in inhibiting the development of neoplasms in epithelial cells of lingual mucosa induced by 4-NQO in the rat.

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Background: Leptin is an adipokine made by fat cells and plays a key role in proliferation, cell survival, migration and immune response. Several studies have suggested that individuals with high serum leptin concentrations would increase the risk of breast cancer. G -2548A polymorphism in the leptin gene is located in the promoter region and is associated with the change of leptin serum level. In this study, the association between G -2548A polymorphism in leptin gene and breast cancer susceptibility was investigated. Methods: This case-control study was done on 374 Iranian women. This study was performed from March 2013 to February 2013. Blood samples from 203 women with breast cancer and 171 age (±5)- matched healthy women were collected. Breast cancer patients were selected from Namazi Hospital in Shiraz city. Genomic DNA was extracted from blood samples. The G -2548A polymorphism of leptin gene was determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Data analysis was performed by SPSS version 18. Logistic regression analysis was used for association of breast cancer susceptibility and G -2548A polymorphism of leptin gene. Results: The A allele frequency was 60% in control group and 72% in breast cancer patients. There was a significant association between A allele in -2548 position of leptin gene and breast cancer susceptibility (OR: 1.8, 95% CI: 1.3-2.4, P<0.001). In the reces-sive effect of the A allele (comparison between AA vs. AG+GG), AA genotype in -2548 region of leptin promoter sequence was significantly increased the risk of breast cancer (OR=2.2, 95% CI: 1.5-3.4, P<0.001). Conclusion: It is concluded that A allele in the -2548 promoter region of leptin gene may act as a recessive allele and increase the breast cancer risk.

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Background: Colorectal cancer is a major cause of morbidity and mortality throughout the world, and its treatments include surgery, chemo-radiotherapy. Despite improvements in clinical outcomes of patients with this tumor over the past decades, prognosis remains poor with a 5-year survival rate of <10%. Angiogenesis inhibitor agents have been recently added to the treatment regimen of this disease. In the past two decades, it has been recognized that selective inhibitors of the cyclooxygenase -2 (Cox-2) enzyme result in the regression in the size of colorectal tumor, and one of its reasons is attributed to angiogenesis inhibition. The present study aimed at identifying the molecular pathways of angiogenesis inhibition by celecoxib. Methods: HCT-116, which is one of the cell lines of Colorectal cancer (separated from human colorectal adenocarcinoma) was provided by the National Cell bank of Iran (NCBI) affiliated to Pasteur Institute. It was then cultured in DMEM (high glucose) culture medium containing 10% FBS, and then treated in the active substance of celecoxib at pharmacological concentrations of 50 mM (C50) and 100 mM (C100). Afterwards, RNA was extracted and cDNA was prepared. The oligonucleotide of HIF-1 Alpha gene (angiogenesis initiator) was prepared and the level of HIF-1 alpha gene expression was assessed with a real-time PCR device in three control, C50 and C100 groups. Results: HIF-1 alpha gene expression significantly decreased in the celecoxib treatment group (compared with control group) with the concentration of C100 (P< 0.001), but no change was observed in the concentration of C50. Conclusion: Angiogenesis is a key factor in the carcinogenesis process and FDA today approved bevacizumab as a first-line treatment for patients with metastatic colorectal cancer. The results of this study showed one of the causes of angiogenesis reduction in celecoxib-treated colorectal cancer. According to clinical findings and basic studies, celecoxib will be hopefully used as a first-line therapy along with chemotherapy in the near future in colorectal cancer. The advantages of this treatment method include its low cost and low side effects.

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Background: Medullary thyroid carcinoma (MTC) occurs in both sporadic (75%) and hereditary (25%) forms. The missense mutations of the rearranged during transfection (RET) proto-oncogene in MTC development have been well demonstrated. Several studies have been published that indicate the molecular analysis of RET gene may offer early identification of those patients at high risk to develop MTC and may provide the opportunity for early intervention. The aim of this study was to investigate frequency of G691S/S904S haplotype in MTC patients and their relatives. Methods: From 2004 to 2014, 358 participants were studied, including 213 patients (119 female, 94 male) and 145 their relatives (79 female, 66 male) in cellular and molecular research center of Shahid Beheshti Research Institute for Endocrine Sciences, Tehran, Iran. Genomic DNA was extracted from peripheral blood leucocytes using the standard Salting Out/Proteinase K method. Nucleotide change detection was performed using PCR and direct DNA sequencing methods. The RET mutations and SNPs, sequences were analyzed. Results: According to DNA sequencing results, 189 individuals (119 patients, 70 relatives) had both G691S (rs1799939) missense mutation in exon11 and S904S (rs1800863) synonymous mutation in exon 15 of RET proto-oncogene. The allele frequency of G691S/S904S haplotype was 35.02% in patients and 29.92% in their relatives. Conclusion: The obtained data showed the frequency of G691S/S904S RET gene haplotype among Iranian MTC patients and their relatives. The G691S and S904S nucleotide changes were in complete linkage disequilibrium, so the results were grouped together and referred to as G691S/S904S haplotype. This haplotype are not considered as oncogenic mutations at this time, its functional role should be investigated. Further analysis is needed to demonstrate the association between this haplotype and MTC development.

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Helicobacter pylori (H. pylori) is the causative agent in development of gastroduode-nal diseases, such as chronic atrophic gastritis, peptic ulcers, mucosa associated lym-phoid tissue (MALT) lymphoma, and gastric cancer. H. pylori has been associated with inflammation in cardia, showing the fact that infection with this bacterium could also be a risk factor for gastric cardia cancer. Gastric cancer is the fourth most common cancer worldwide. This is the second leading cause of cancer-related deaths, and ap-proximately 700,000 people succumb each year to gastric adenocarcinoma. It has been estimated that 69% of the Iranian population currently harbor H. pylori infection. The prevalence of duodenal ulcer and gastric cancer is high in Iranian populations. However, this has been largely influenced by geographic and/or ethnic origin. Epidemi-ology studies have shown that host, environmental, and bacterial factors determine the outcome of H. pylori infection. The bacterium contains allelic diversity and high genet-ic variability into core- and virulence-genes and that this diversity is geographically and ethnically structured. The genetic diversity within H. pylori is greater than within most other bacteria, and its diversity is more than 50-fold higher than that of human DNA. The maintenance of high diversification makes this bacterium to cope with particular challenges in individual hosts. It has been reported that the recombination contributed to the creation of new genes and gene family. Furthermore, the microevolution in cagA and vacA genes is a common event, leading to a change in the virulence phenotype. These factors contribute to the bacterial survival in acidic conditions in stomach and protect it from host immune system, causing tissue damage and clinical disease. In this review article, we discussed the correlation between H. pylori virulence factors and clin-ical outcomes, microevolution of H. pylori virulence genes in a single host, microevolu-tion of H. pylori during primary infection and progression of atrophic gastritis to ade-nocarcinoma, and H. pylori infection status in Iran. Finally, we put forward the hy-pothesis that if the pattern of nucleotide sequence evolution shifts from recombination (r) to mutation (m) and the r/m ratio is reduced, bacterial pathogenicity may be re-duced while maintaining the bacterial life. However, this hypothesis should be further studied with future experiments.

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Background: Endometrial carcinoma is considered the most common gynecological cancer in the world. Pelvic and para-aortic lymphadenectomy is widely advised based on FIGO staging system. Intra-operative frozen sections analysis is used to identify pa-tients at high risk for pelvic and para-aortic nodal metastasis evading lymphadenec-tomy in low-risk patients. However there is still some controversy concerning the effi-ciency of IFS. The aim of this study was to determine the validity and precision of fro-zen section diagnosis and gross examination of uterine specimen compared to the final histological results in patients with endometrial cancer. Methods: Patients diagnosed as endometrial cancer based on office biopsy using a Pipelle or D&C who underwent surgical staging were compared for frozen section anal-ysis and permanent diagnosis. Patients with the history of radiotherapy or other types of cancer or co existence malignancies were excluded. Results: There was no relation between the tumor size and lymph node involvement and the results were not significant (P= 0.1). Frozen section analysis was significantly accurate and correct in predicting final histopathological results (P< 0.0001). It has been shown that in more than 90% of patients the diagnosis made by frozen section analysis was in accordance with final pathology with considerable sensitivity and spec-ificity. Gross examination was also precise in determining myometrial microscopic in-vasions (P< 0.0001). Conclusion: Although the sample size of the studied population was small but our study results support the previous data and suggest that in early stages and low grade tumors, gross examination and frozen section diagnosis are conveniently predictive of lymph node metastasis. These data might be useful for prediction of tumor invasion using frozen section and gross examination in low grade tumors and early stages and for doing complete surgical staging and lymph node sampling. However the im-portance of surgical staging always must be considered in patients who need systemat-ic lymphadenectomy. In overall these data might help to come up with new guidelines for surgical risk assessment in endometrial cancer.

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Background: Helicobacter pylori vacA (vacuolating toxin A) gene is comprised of mid- (m), intermediate- (i) and signal-regions. Recently, the vacA-i region genotype has been suggested to be a better predictor of disease severity than either the s- or m-region. The main aim of the present study was to determine the associations of i region poly-morphisms of vacA gene with gastric cancer (GC) and peptic ulcer disease (PUD) in Azerbaijan Province patients. Methods: A number of 89 patients were enrolled. The biopsy samples were taken from patients referring to the endoscopy units of Imam Reza and Shahid Madani Hospitals, Tabriz, Iran from August 2012 to May 2013. The genotype frequencies of vacA-i1 and i2 in were studied using polymerase chain reaction (PCR). Results: The frequency of vacA-i1 and i2 was 51.68% and 48.31%, respectively. The genotypic frequency of vacA-i1 in patients with GC (21/24, 87.5%) was significantly higher than in those with non-atrophic gastritis, NAG (19/48, 39.58%). In contrast, the genotypic frequency of vacA-i2 in patients with NAG, PUD, and GC was 60.42%, 64.70%, and 14.28%, respectively. The results of multiple linear and logistic regression analyses confirmed the intensity of correlation of vacA-i1 allele with GC compared with control group (NAG). No significant correlation was found between the vacA-i-region alleles and PUD risk. Conclusion: We have proposed that the H. pylori vacA-i1 genotype could be an im-portant biomarker for predicting the gastric cancer risk in Azerbaijan Province in Iran. However, due to the difference in the allelic frequency of this gene in H. pylori strains from different parts of the world, the vacA-i1 genotype usefulness in predicting the gas-trointestinal diseases is dependent to the geographic origin of the strains.

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Background: Biosynthesis of nanoparticles has attracted the attention of the scientific community in nanotechnology and biotechnology due to their extensive application in the area of material sciences and medicine. Nowadays, despite a various application of nanomaterial’s, there is a little information about their impact on human health. In this study, we investigated the comparative study on cytotoxicity effect of biological and commercial synthesized nanosilver on human gastric carcinoma (AGS) and normal lung fibroblast (MRC-5) cell lines. Methods: The current experimental study was carried out in Islamic Azad University, East Tehran Branch, from April to November 2014. The biological synthesis of nanosilver was obtained from Eucalyptus plant extract as a reducing agent. Further to more analysis, morphological study on size and shape of developed biological nanosilver was characterized by performing scanning electron microscopy and dynamic light scattering. AGS and MCR-5 cell lines were treated with various concentration of nanosilver for 24, 48 and 72 hours. Finally, the cell viability was evaluated by using MTT assay. Results: The results show that the nanosilver exerts a dose-dependent inhibitory effect on viability of cells. At 100µg/mL of commercial and biological synthesized nanosilver, the viability of AGS was reduced to 7.47±0.002% (P=0.002) and 3.65±0.01% (P=0.003) after 72 hours, respectively. In addition, the viability of MRC-5 at the same condition was reduced to 10.27±0.19% (P=0.001) and 9.16±1.53% (P=0.002), respectively. Conclusion: Based on a thorough literature surveys, the present study is the first research about biosynthesis of nanosilver using Eucalyptus plant extract. This eco-friendly and cost effective method can be used for large scale production of silver nanoparticle. In addition, based on the current obtained data, commercial and biological synthesized nanosilver can more inhibitory effect on cancer cells compared to the normal cells. Hence, silver nanoparticles might be used as a new strategy for treating many human cancers. However, further studies are necessary to ascertain their potential as anticancer agents.

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Background: Metastasis associated miRNA (metastamiR) opened a new field of anti-metastatic therapy which have a great potential of treatment for the most lethal aspect of cancer, metastasis. The pleiotropic nature of gene regulation exhibited by certain miRNAs that showed that miRNAs might be endowed with a capacity to function as crucial modulators of tumor metastasis. MiR-31 is a pleiotropic anti-metastatic miRNA whose expression decreased significantly in metastatic breast cancer cells. MiR-31 has multiple roles in metastasis cascade. Therefore, using the miR-31-restoration based therapy could be an efficient anti-metastatic strategy for cancer therapy. Methods: This research was performed from May 2014 to October 2015 in Tarbiat Modares University in Tehran, Iran. The double-strand oligo of mature miR-31 was cloned into pcDNA 6.2gw/EmGFP according to the manufacturer instruction. The MDA-MB231, MCF-7 breast cancer cell lines were cultured and their miRNAs have been extracted. The expression of miR-31 has been quantified by Real time-PCR be-fore transfection of construct contained miR-31 into two cell lines and in normal breast cells. Then the constructs contain miR-31 have been transfected in to two cell lines. The expression of miR-31 has been quantified after 48 hours. Scratch and invasion as-say have been carried out for assessing the level of migration and invasion. Results: The result of Real time-PCR before transfection of constructs contained miR-31 have been shown 4 fold and more than 100 fold reduction in expression of miR-31 in MCF-7 and MDA-MB231 respectively in comparison to miR-31 expression in nor-mal breast cells, but after transfection of miR-31 construct to MDA-MB231 the quan-tification of expression showed the significant increase in mir-31 expression and 20 fold reduction in invasive and 10 fold reduction in migratory characteristics of MDA-MB231 in comparison to MCF-7. Conclusion: Metastasis associated miRNA have been represented a promising candi-dates in the field of anti-metastatic therapy and miR-31 as a powerful member of this family can function very effectively in order to inhibit the metastasis and introduce the new possibility of metastasis inhibition.

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Background: The molecular studies indicate some of the genes in the promoter region itself, will undergo methylation. Methylation of CpG islands in the promoter region of that cause silence or reduced expression of genes involved in cell growth pathways, which are colorectal cancer causing agents. Detection of methylation status can be used as a marker for cancer diagnosis and prediction of disease. CDKN2A tumor suppressor gene encodes a protein, which inhibit CDK 4/6 and loss of retinoblastoma protein phosphorylation (pRb) is involved. The purpose of this study was to investigate the molecular hypermethylation in exon 1 of CDKN2A gene in patients with colorectal cancer and normal subjects. Methods: In this case-control study, the study population consisted of 20 patients with colorectal cancer and 10 healthy persons. Samples in paraffin blocks were prepared in pathology department of Mehr Hospital, Tehran, Iran, from December 2010 to June 2012. Then, specific primers were designed for the methylation and Non-methylation of CDKN2A gene. To determine the level of exon 1 methylation of CDKN2A gene, methylation-specific polymerase chain reaction (MSP) method was performed. Results: In this study, hypermethylation in exon 1 of CDKN2A gene were observed in 80% of tumor tissues (16 cases) and 20% of normal tissues (2 cases). The patients aged older than 50 years, had a higher CDKN2A gene methylation and frequency than patients younger than 50 years old (66% vs 34%) (P<0/001). Conclusion: The result of this study has been confirmed the role of CDKN2A gene promoter methylation of CpG sites of colorectal cancer as the leading cause of colorectal cancer. These data suggest that epigenetic silencing via aberrant methylation of the CDKN2A promoter plays a critical role in the inactivation of this tumor suppressor gene in colorectal cancer and can be used as a marker for early detection and identification of potential applications.

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Cancer/Testis antigens (CTAs) as a group of tumor antigens are the novel subjects for developing cancer vaccine and immunotherapy approaches. They aberrantly express in tumors with highest normal expression in testis, and limited or no expression in normal tissues. There are important similarities between the processes of germ-cell and cancer cell development Spermatogenesis begins at puberty when expression of novel cell-surface antigens occurs when the immune system has been refined the ability to distinguish self from non-self. Whereas macrophage and lymphocytes are commonly found within interstitial spaces of the testis, these antigen-presenting cells are rarely seen within the seminiferous tubules. These observations have led to the concept of the immune privileged site for testis. Localized normal expression of the CT genes in testis that makes them immunogenic for immune system, in one side, and their abnormal expression in different kinds of cancer cells, in the other side, has make them as promising target for developing cancer vaccines and new cancer therapeutics approaches. In malignancies, gene regulation is disrupted which results aberrant expression of CT antigen in a proportion of tumors of various types. For some CTAs, data support their fundamental role in tumorigenesis. Several authors believe it is not clear whether they have an essential role in tumorigenesis or they are by-products of chromatin variations in cancer. There is a growing list of CTAs within them advanced clinical trials are running by using some of them in cancers like lung cancer, malignant melanoma and neuroblastoma. In this review we discuss the gene TSGA10 as an example of CT genes. TSGA10 expresses in its highest levels in elongating spermatids and localized in the fibrous sheath of mature sperm. This gene is proposed as a serological biomarker in cutaneous lymphoma. Its abnormal expression has been reported in different cancers such as acute lymphoblastic leukemia, breast, brain, gastrointestinal and a range of other cancers either in mRNA or protein levels. It has an important role in angiogenesis in cancer tumors because of its effects in the gene hypoxia-inducible factor (HIF1). Absence or lack of TSGA10 expression has been reported in ascosporic infertile men.

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Background: Prostate-Specific Antigen (PSA), also known as gamma-seminoprotein or kallikrein-3 (KLK3), is the best marker for early diagnosis of prostate cancer. Since age and race are affecting PSA levels, determining age-specific reference ranges of PSA in every community is necessary for increasing the efficiency rate of PSA. The aim of the present study was to evaluate the normal distribution of total prostate-specific antigen (TPSA) and free prostate-specific antigen (FPSA) and determine age-specific reference ranges of PSA in Iranian men. Methods: In this cross-sectional study, 1200 normal men with the age range of 50 to 79 referred to Shahid Rajaie Hospital, Qazvin Province in Iran, from 2011 to 2013. After excluding patients with prostate cancer and urinary tract infection, 1020 men were included in this study. Then, their blood samples were collected and after the extraction of serum from blood, serum levels of FPSA and TPSA were measured using commercial kits the reference range of PSA was specified for each age group and compared with reference ranges of other populations. Results: The mean age of the patients was 61.03±7.5 years and the mean values of FPSA and TPSA were 0.47±0.6 ng/ml and 1.56±2.05 ng/ml, respectively. PSA serum levels (95th percentile range) in 50 to 59, 60 to 69 and 70 to 79-year age groups were 0-3.6 ng/ml, 0-5.7 ng/ml and 0-6.8 ng/ml, respectively. TPSA (r= 0.2, P< 0.001) and FPSA (r= 0.22, P< 0.001) were significantly associated with age. In addition, a significant relationship was found between TPSA serum levels and alcohol consumption (P= 0.017), smoking (P< 0.001) and family history of prostate cancer (P= 0.014). Conclusion: Findings of the present study showed that PSA levels are correlated with age. It was also revealed that the PSA age-specific reference range obtained in this study is different from other races and is specific to Iranian men. Therefore, age-specific reference ranges of PSA obtained in the present study can increase PSA test sensitivity and specificity by reducing unnecessary diagnostic procedures and early detection of prostate cancer in Iranian men.