Association of IL-10, IL-18, and IL-33 genetic polymorphisms with recurrent pregnancy loss risk in Iranian women.

Recurrent pregnancy loss (RPL) is a heterogeneous disease with three or more consecutive abortions before 20 weeks of pregnancy. Recently, inflammatory factors such as interleukins (IL) have been found to be a significant factor in the RPL. The objective of this study was to investigate the association between RPL and IL-10 (rs1800896), IL-18 (rs1946518) and IL-33 (rs1929992) genes polymorphisms in Iranian women. The study participants consisted of 300 women with RPL and the control group comprised of 300 healthy women with successful delivery. Genomic DNA was extracted from peripheral blood, and genotyping was performed by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP). There were no significant differences in the frequencies of genotype and allele in IL-10 gene polymorphism (rs1800896) between patients and control group (p > .005). In contrast, there were significant differences in the frequencies of CC genotype in IL-18 gene polymorphism (rs1946518) between patients and the control groups (p = .004; OR =0.990; 95% CI: 0.320-8.855). Also, there were significant differences in the frequencies of GA genotype in IL-33 gene polymorphism (rs1929992) between patients and the control groups (p = .001; OR =0.955; 95% CI: 0.239-9.807). Present study showed that the rs1800896 polymorphism (IL-10) might not play role in RPL in the Iranian population; whereas rs1946518 (IL-18) and rs1929992 (IL-33) polymorphisms may be associated with the risk of RPL in the Iranian women.

In vivo and in vitro impact of miR-31 and miR-143 on the suppression of metastasis and invasion in breast cancer.

PURPOSE: The microRNA (miR)-31 and miR-143 are pleiotropic anti-metastatic miRs, with an expression that decreases significantly in metastatic breast cancer cells. The aim of this study was to investigate the effect of miR-31 and miR-143 inhibition on metastasis and invasion in both MDA-MB231, MDA-MB468 as well as the MCF-7 breast cancer cell lines and 5-week old female mice. METHODS: Following the cloning of miR-31 and miR-143 into vectors, their expressions were determined before treatment with constructs of miR-31 and miR-143 in cancer cell lines and normal breast cells. Then miR-31 and miR-143 were transfected to the cell lines and the expression was assessed after 48 hrs. Moreover, the levels of migration and invasion were determined in cell lines. These experiments were performed in 5-week old female mice. RESULTS: The results showed that miR-31 expression before the transfection of miR-31 construct was decreased 4, 70 and 100 times in MCF-7, MDA-MB468 and MDA-MB231 cell lines, respectively, in comparison to normal breast cells; but after the transfection of miR-31 construct, the expression of miR-31 increased 80 times. Additionally, invasion and migration decreased by 15 and 10 times in MDAMB-468. All of the modifications in miR-143 were low in comparison to miR-31. The results of the in vivo experiments were approximately the same as in the in vitro experiments. CONCLUSIONS: It appears that the use of miR-31 is highly efficient than miR-143 in the inhibition of invasion and metastasis in breast cancer. Our study improved our conception about miR-31 and miR-143 and their roles in the identification and therapy of breast cancer.

Common rs5918 (PlA1/A2) polymorphism in the ITGB3 gene and risk of coronary artery disease.

INTRODUCTION: The T to C transition at nucleotide 1565 of the human glycoprotein IIIa (ITGB3) gene represents a genetic polymorphism (PlA1/A2) that can influence both platelet activation and aggregation and that has been associated with many types of disease. Here, we present a newly designed multiplex tetra-primer amplification refractory mutation system - polymerase chain reaction (T-ARMS-PCR) for genotyping a single nucleotide polymorphism (SNP) (dbSNP ID: rs5918) in the human ITGB3 gene. MATERIAL AND METHODS: We set up T-ARMS-PCR for the rs5918 SNP in a single-step PCR and the results were validated by the PCR-RFLP method in 132 coronary artery disease (CAD) patients and 122 unrelated healthy individuals. RESULTS: Full accordance was found for genotype determination by the PCR-RFLP method. The multiple logistic regression analysis showed a significant association of the rs5918 polymorphism and CAD according to dominant and recessive models (dominant model OR: 2.40, 95% CI: 1.33-4.35; p = 0.003, recessive model OR: 4.71, 95% CI: 1.32-16.80; p = 0.0067). CONCLUSIONS: Our T-ARMS-PCR in comparison with RFLP and allele-specific PCR is more advantageous because this PCR method allows the evaluation of both the wild type and the mutant allele in the same tube. Our results suggest that the rs5918 (PlA1/A2) polymorphism in the ITGB3 gene may contribute to the susceptibility of sporadic Iranian coronary artery disease (CAD) patients.