ABSTRACT
Multiple sclerosis (MS) is a chronic infl ammatory demyelinating disease of the central nervous system. Interleukin-2 (IL-2) is identifi ed as the crucial and main immunoregulatory cytokines. Previously, we showed signifi cant association between -330 T/T IL-2 genotype and relapsing remitting MS among Iranian population. In this study we investigated 100 relapsing remitting, 30 secondary progressive MS and 125 healthy controls to compare the relapsing remitting and secondary progressive course MS in association to -330 IL-2 polymorphism. Our results showed that the -330 T/T IL-2 genotype was signifi cantly more frequent in relapsing remitting and secondary progressive MS than controls. The signifi cant increased frequency of -330 T/T IL-2 genotype in secondary progressive than relapsing remitting MS, imply -330 T/T IL-2 genotype can cause higher susceptibility to secondary progressive MS than relapsing remitting.
ABSTRACT
Multiple sclerosis [MS] is a chronic autoimmune disease due to demyelination of the central nervous system. It is believed that cytokines are involved in the pathogenesis of MS. The interleukin-2 [IL2] gene is powerful functional candidate that is involved in immune regulation and operation. In this study, for the first time, we investigated the effect of -475 A/T and -631 G/A IL2 polymorphisms on MS disease in Iranian patients. In this case-control study, 100 MS patients [mean age: 32.95 +/- 6.51 years, age range: 20-42 years] selected according to McDonald criteria, and 100 ethnically, sex and age matched healthy controls [mean age: 29 +/- 7.8 years, age range: 20-52 years] with no personal or family history of autoimmune diseases were studied. The restriction fragment length polymorphism-polymerase chain reaction [RFLP-PCR] method was applied to define different alleles and genotypes of IL2 promoter single nucleotide polymorphism -475 A/T as well as -631 G/A among individuals. chi [2] was calculated and Fisher's exact test was applied to analyze the obtained data. The value of p <0.05 was considered significantly. Evaluation of the -475 IL2 revealed that T allele and A/T genotype are present in 2% and 4% of MS patients, respectively, whereas T allele was absent in control samples. The comparison between alleles and genotypes in MS patients and healthy controls was not significant [p=0.1]. For the -631 position, 1% and 2% of MS patients carried A allele and A/G heterozygote genotypes, respectively. All control samples had G allele and G/G genotype. The differences between patients and controls were not significant [p=0.4]. Moreover, our results showed a very low frequency of T at -475 and A at -631 IL2 position in each of the two groups. Both -475 and -631 IL2 polymorphisms were higher in MS patients as compared to controls, but the frequency differences were not significant. Based on these data, it is suggested that the -475 and -631 IL2 polymorphisms as functional promoter position may be involved in IL2 expression and regulation. To find out the exact effect of the mentioned SNPs on susceptibility to MS, study on a larger sample size is suggested
Subject(s)
Humans , Female , Male , Interleukin-2 , Polymorphism, Genetic , Polymorphism, Single Nucleotide , Case-Control StudiesABSTRACT
Functional defects in mitochondria are involved in the induction of cell death in cancer cells. The process of programmed cell death may occur through the mechanisms of apoptosis. Several potential lead molecules such as Camptothecin [CPT] and its analogues have been isolated from plants with anticancer effect. The aim of the present study was to understand the necrotic effect versus apoptotic nature of CPT in HeLa cancer cells. The antiproliferative activity of CPT was estimated through 3-[4, 5- Dimethyl thiazol-2-yl]-2, 5-diphenyl tetrazolium bromide [MTT] assay and DNA fragmentation analysis using gel electrophoresis. Lactate Dehydrogenase [LDH] activity and cell morphology were assessed under control and CPT exposed conditions to evaluate the necrotic effect of CPT. The results showed that CPT inhibited the proliferation of HeLa cells in a dose-dependent manner with an Inhibitory Concentration 50% [IC50] of 0.08 +/- 0.012 microg/ml. However the significant [P<0.05] increase happens in LDH activity at concentrations 1x10-1microg/ml and above. Morphological changes showed that CPT in low concentrations induced an apoptotic mechanism of cell death, such as cell shrinkage and characteristic rounding of dying cells, while at high concentrations showed necrosis changes. The characteristic DNA ladder formation of CPT-treated cells in agarose gel electrophoresis confirmed the results obtained by light microscopy and LDH assay. Camptothecin as an anticancer drug may have antiproliferative effect on HeLa cancer cells in low concentrations, through its nature of induction of apoptosis. The border line between necrotic effect and apoptotic nature of CPT in HeLa cancer cells has been found to be at concentration of 1x10-1 microg/ml