effect of 5'untranslated region polymorphism in EGF gene, rs4444903, on colorectal cancer

The purpose of this study was to determine the relationship of rs4444903 [EGF+61A/G] SNP genotype with colorectal cancer and tumor stage in an Iranian population. Epidermal growth factor [EGF] is one of the important proteins that determine survival of cells. EGF binds to its receptor on the cell surface and then activates some of the cell signaling pathway networks within cells that lead to activation or deactivation of factors which are responsible for growth and apoptosis of cells. In this study we assessed the association in EGF polymorphism rs4444903 with colorectal cancer [CRC] in Iranian population. We conducted case-control study to investigate the association of polymorphism rs4444903 in EGF, with colorectal cancer risk in Iranian population. Analyzed Polymorphism of EGF rs4444903 with restriction fragment length polymorphisms [RFLP] among two groups of subjects consisting of including 220 cases with colorectal cancer and 220 healthy individuals as controls. Mutations were confirmed in 10% of the samples by direct sequencing. The frequencies of AA, AG and GG genotypes among cases with colorectal cancer were 28.2, 46.8, and 25.0% respectively and in controls genotype frequencies were 23.2, 56.4, and 25.0%, respectively. Frequency of A allele among case group was 51.6% and for control group was 51.4%. The frequency of G allele in case and control was, respectively 48.4% and 48.6% [OR= 1.009, 95% CI= 0.775-1.315; P= 0.946]. The percentage of Stage 0, I, II, III, IV were 5%, 9.35%, 38.84%, 30.21% and 16.54%, respectively, among the cases. However, no significant association between this polymorphism and CRC stage was observed [p=0.626]. Our data suggest a SNP rs4444903 may not represent a risk factor in the development and progression of CRC among Iranian population

Assesment of FLT3-gene mutations among children with acute leukemia

FLT3-gene mutations cause leukemic cells to proliferate uncontrollably and leads to a poor prognosis. The aim of this study was to explore appropriate at diagnostic molecular tests and to screen mutations that occur in patients with acute leukemia. In this basic study, 91 children with acute myeloid leukemia [AML] and acute lymphoid leukemia [ALL] were investigated for FLT3-gene mutations, including ITD mutation [Internal Tandem Duplication] and the point mutation that is coded by exon 17. ITD mutation in FLT3 receptor was analyzed by PCR [Polymerase chain reaction] in 11, 12 exons and 11 intron using designed primers. For analysis of point mutation of Exon 17 in FLT3 receptor gene, the genomic DNA of patient was amplified using the PCR. Resulted PCR products were studied by ECORV enzyme and restriction length polymorphism [RFLP]. In cases of positive ITD, the sequencing method was applied. Of 91 acute leukemia patients, ITD mutation was observed in 7 cases. Two of 91 patients had point mutation of D835. Distribution of ITD and point mutation of D835 mutation was not identical in FAB subtypes. FLT3-gene mutations are prevalent mutation in children with acute leukemia. So, it can be decided about the treatment after molecular diagnosis of this mutaions, independent of FAB classification and before the treatment get started

Evaluation of relation between mutant VirB2 virulence factor in Brucella melitensis with immunogenicity and intracellular survival

The Brucella melitensis virB operon, encoding a type IV secretion system [T4SS], is required for intracellular replication and persistent infection in the mouse model. The product of the second gene of the virB operon, virB2, is predicted to be localized at the bacterial surface, where they could potentially interact with host cells. Studies to date have focused on characterization of transposon mutations in this gene, which is expected to exert polar effects on downstream genes. We researched on the evaluation of relation between virB2 mutant with immunogenicity in mouse model and intracellular replication in macrophages J774. In order to determine whether VirB2 is required for the function of the T4SS apparatus, we constructed and characterized deletion mutation of virB2 and kanamycin resistance gene replaced instead of virB2. For demonstration of intracellular replication, macrophages J774 and BALB/c mices were infected with wild type Brucella melitensis and mutated. After 48 h, number of mutated Brucella severe decreased severly compaired to wild type in macrophages J774, and Brucella with virB2 deletion decreased from 1x10[6] CFU/spleen less than 1000 CFU/spleen during 8 weeks, also total IgG increased in both but IL-12 and IFN-gamma increased only in wild type. VirB2 was essential for intracellular replication in mouse models and J774 macrophages. The virB2 mutant was unable to cause persistent infection in the mouse model, demonstrating the essential role of VirB2 in the function of the T4SS apparatus during infection and secreting IL-12 and IFN-gamma, but VirB2 dispensable for secretion of total IgG