RESUMEN
Background: Cervical cancer is the second most common female cancer worldwide. Inhibitors of apoptosis proteins [IAPs] block apoptosis; therefore, therapeutic strategies targeting IAPs have attracted the interest of researchers in recent years. Apollon, a member of IAPs, inhibits apoptosis and cell death. RNA interference is a pathway in which small interfering RNA [siRNA] or shRNA [short hairpin RNA] inactivates the expression of target genes. The purpose of this study was to determine the effect of constructed shRNAs on apoptosis and growth inhibition through the suppression of apollon mRNA in HeLa cell line
Methods: Three shRNAs with binding ability to three different target sites of the first region of apollon gene were designed and cloned in pRNAin-H1.2/Neo vector. shRNA plasmids were then transfected in HeLa cells using electroporation. Down-regulation effects of apollon and the viability of HeLa cells were analyzed by RT-PCR, lactate dehydrogenase assay, and MTT assay, respectively. Also, the induction and morphological markers of apoptosis were evaluated by caspase assay and immunocytochemistry method
Results: The expression of shRNA in HeLa cells caused a significant decrease in the level of apollon mRNA1. In addition, shRNA1 effectively increased the mRNA level of Smac [as the antagonist of apollon], reduced the viability of HeLa cells and exhibited immunocytochemical apoptotic markers in this cell line
Conclusion: Apollon gene silencing can induce apoptosis and growth impairment in HeLa cells. In this regard, apollon can be considered a candidate therapeutic target in HeLa cells as a positive human papillomavirus cancer cell line
RESUMEN
In thalassemic children, HBV infection is common, thus immunization against will reduce and prevent the rate of infection. The aim of this study was to evaluate the efficacy of HBV immunization and the prevalence of HBV infection in beta-thalassemic children in Tehran. To assess the efficacy of immunization and determine the immune response of children with beta-thalassemia, sera of 99 children who had received three doses [10/20 micro g] of recombinant HBV vaccine in months 0, 1, 6, were selected and tested for HBsAg, HBsAb and anti-HBc by ELISA method. Also, these sera were tested for HBV DNA using nested-PCR method. In 99 beta-thalassemic children, 89 [89.9%] were anti-HBs positive [responders] and 10 [10.1%] anti-HBs negative [non-responders]. 3 [3.03%] were anti-HBc positive and 1[1.01%] was HBsAg positive. HBV DNA was not detected in any of them. Our results have revealed that hepatitis B vaccine is highly immunogenic for thalassemic children and particularly well tolerated