investigation of the relation between the poly morphism c14 orf101 and mirna-124 genes in the development of gastric cancer in khorasan razavi province

Gastric cancer in the world is one of the most common malignancies in humans and is the second most common cause of cancer death. According to the World Health Organization (WHO),in 2012, around one million people in the world have been diagnosed with gastric cancer, of which about 70% live in developing countries, especially in East Asia, accounting for 6.8% of all cancers Make it look. About 50% of gastric cancers are malignant. There are also 723,000 deaths from gastric cancer every year, equivalent to 8.7% of all deaths. If cellophane cancer is considered as a multifactorial process, the parallel effects of the environment and genetics play an important role. The same collaborative process has led to extensive studies on genetic changes, as is the case with most malignancies, both genetic factors and the environment play an important role in the pathogenesis of gastric cancer. Considering the prevalence of gastric cancer in Khorasan Razavi province and the genetic differences of people in different countries, this study will be carried out for the first time in Iran. It is hoped that its results will be effective in the treatment and prevention of the disease. This study was performed on 50 gastric cancer patients and 50 healthy controls. In this study, genomic DNA was extracted from paraffin blocks in gastric cancer patients. Then genotyping of C14 orf101 and mir- 124 genes was performed using PCR-RFLP and statistical analysis using Medcalc software. Gastric cancer is the most common cancer of the digestive tract, especially in the northwest of the country. In Iran, unlike in western countries and Japan, the incidence of gastric cancer has been increasing over the past two decades (2). There has been some progress in our understanding of genetic variations in the incidence of gastric cancer, which is mostly in the type of intestine. The gene that focuses most on the suppressor gene is the p53 tumor. The mutation in this gene has a high prevalence in gastric cancer (38-71%). The mutation in the p53 gene is 38% in the intestinal metaplasia and 58% in the gastric dysplasia, and perhaps the mutation in the p53 gene is an early occurrence in the pathogenesis of gastric cancer. In studies on mice with p53 homozygous mice, it was observed that these mice exhibited a growing response to Helicobacter pylori infection in comparison with wild mice. Generally, this growing proliferation is associated with an increased risk of gastric cancer. Of course, the absence of p53 does not indicate a worse prognosis in patients with gastric cancer (intestinal type). Looking at all these issues, it is predicted that p53 gene inactivation is important in the early pathogenesis of gastric cancer (15). Examination of the genes discussed in the thesis The other name for the gene is the (TMR260) C14 orF101. The product of this gene is a transmembrane protein. TMEM260 is located on the short arm of chromosome 14. The size of this miRNA is 4.278 bp. After examining the amino acid sequence, its similarity is between 73% and 86% with other organisms, such as birds and fish, and other rules (12). The gene number rs4901706 is in NCBI (15). Other RSs of the C14 orF101 gene Which are as follows: (GG vs.AG + AA) rs 4901704, Rs1044129 (AA rs AGTGG), Rs11337 (GG rs GT, TT)Rs3660 (GG vs CG vs. CG.CC), Rs1053667 (TT vs. CT + CC), Another result of this study is that SNP rs 4901706 present in the C14 orF101 is a marker for predicting the risk of gastric cancer (10). Gene MIR-124 On the short arm of chromosome 8, the number of this gene in the NCBI is rs531564 MicroRNA processor, MIR-124 is an uncoded RNA molecule that has been identified in type MI0000373 (1) in type MI0000302 (2) nematodes and worms, in MI0000150 type mice and in MI0000443 (3) human species. . Mature 124 microRNAs contain 21 nucleotides that are made up of the arm or branch of the hairpin sequence of the Dicer enzyme (disperse). In many studies, the expression of MIR-124 has been altered, but this change does not affect the differentiation of the neuronal cells and affects the types of cancers, especially gastrointestinal cancers, and in particular the stomach (4). But some other studies have shown that MIR-124 has an effect on the differentiation of neuronal cells (5). Studies have shown that the target of MIR-124 - the micro-RNA of the antioperative proteins of the neurons Sep (C-end of the end phosphatase 1) (6) or directly on the PTBP / MRNA effect as a major and general coding agent, Pre-MRN in non-neuronal action cells (7) and MIR-124 prevent changes in glutamate dischargers in the anterior (frontal) cortex of the cerebral brain and impairment of brain function (8). The easiest thing about the genes mentioned is that the enzymes of this gene are completely specific, and in a study done on the Gene Runner-NCBI-Neb cutters sites, luckily, a specific enzyme was found for the hygenic process, which is discussed in full in the next discussion. 3- 3 Sampling and specification This case-control study was designed to investigate the relationship between miRNA polymorphisms in unregistered region 3 & apos; C14 orf101 and mir-124 genes in gastric cancer in Khorasan Razavi province, Ghayem Hospital of Mashhad. Initially, according to previous studies and consultation with a statistician, samples from the pathology department of Ghayem Hospital related to 1395.50 paraffin blocks with gastric adenocarcinoma (5 women and 45 males aged 65-65), based on endoscopic results Pathology ((Intestinal adenocarcinoma-moderally diff under the supervision of the master of the jinn 4 Extracting genomic DNA from paraffin tissue DNA extraction was carried out according to the following instructions according to the kit instructions: One milliliter of xylene was poured into each microtip and ten seconds later, the vortex was started for 5 minutes centrifugal at about 12000 rpm. Paraffin depletion of the paraffin block was repeated 3 times. Take out the topical solution and then use a total alcoholic 96 and 70 to make the texture. Add 1cc of absolute ethanol and centrifuge it again with the previous round of the previous day, and then do it with alcohol 96 and 70 and finally with water. The distilled water was distilled twice a day at a rate of 1cc 12,000 centrifuges. Empty the supernatant, with the tip of the sampler, the bottom of which there is no alcohol. Then we transfer the tissues that are left in the remaining tubes to another sterile tube. 3- 5. Extracting DNA To this end, the Pars Tusar DNA extraction kit was used with Cat No.A101211. First, before the DNA was extracted, the required solutions of PW1, PW2, PX, PL, PTB were made according to the kit kit and kept at the recommended temperature. The following measures are taken to prepare the main solutions 100 μl of absolute alcohol was added to PW1 material (Wash 1) To prepare the PW2 solution, 48 μl of absolute alcohol was added to PW2. (Wash 2) To prepare a protein kinase solution K, the specific kinase solvent contained in the kit was added to the powdery polysulphide and kept at -20 ° C. The tissue is ready for paraffin dehydration Add 1 to 200 μl of PL solution to the microtubule, and then twenty lanta proteins K protamine K, then place in the microtubes and then 15 seconds in 56 ° C for 2 to 3 hours. 200-2000 lanta pure ethanol 96 g. Mix it and mix it with a microtubule on a column of DNA (colon) with a sample of the inside of the column. Then centrifuge 1 minute about 13000 We remove the column and remove the microtubule below it. New microtubes. After placing the column on the new microtubule, we will put the column back to the next microtome.Then, again, we will discard the microtubule again for 1 min 13,000. 4-dimensional new microtubule.Again, 700 ml of the solution of 2 will be added to the column. Again, 1 min 13000 microtitre centrifuge will be removed, then the column will be kept. 5-Repeat the previous step. Repeat the 500 Landavaw 2 for 1 minute at 13,000. Then remove the microtubule again. The last step is to add Ellution (PE) 50 to produce a more concentrated DNA. 6 pillars on microtubes. 50 lands of PE (Ellution) remain at room temperature for 3 minutes, and then drop to 3 minutes at room temperature. 7. Then, 1 minute in 13000 centrifuges, remove the amniotic membrane. Next, the microtubule is the remaining DNA. After the completion of the work, put the agent into the freezer 20- until the next test and carryout the PCR. Evaluation of DNA quality - Primers used In the PCR reaction of the desired gene, two specific primers suitable for propagation of the genetic range containing the polymorphic componentwere used, the characteristics of which are shown in Table (3-4). In order to identify alleles and genotypes, PCR-RFLP technique was used. The primers were synthesized by South Korea s Bioneercompany.

Keywords:

Gastric Cancer , Stomach Cancer , Gene and Cancer , Cancer Genetics