Résumé
Background: Cancer/Testis Antigens [CTAs] are a subgroup of tumor-associated antigens which are expressed normally in germ line cells and trophoblast, and aberrantly in a variety of malignancies. One of the most important CTAs is Developmental Pluripotency Associated-2[DPPA2] with unknown biological function. Considering the importance of DPPA2 in developmental events and cancer, preparing a suitable platform to analyze DPPA2 roles in the cells seems to be necessary
Methods: In this study, the coding sequence of DPPA2 gene was amplified and cloned into the retroviral expression vector to produce recombinant retrovirus. The viral particles were transducted to Esophageal Squamous Cell Carcinoma [ESCC] cell line [KYSE-30 cells] and the stable transducted cells were confirmed for ectopic expression of DPPA2 gene by real-time PCR
Results: According to the critical characteristics of retroviral expression system such as stable and long time expression of interested gene and also being safe due to deletion of retroviral pathogenic genes, this system was used to induce expression of DPPA2 gene and a valuable platform to analyze its biological function was prepared. Transduction results clearly showed efficient overexpression of the gene in target cells in protein level due to high level of GFP expression
Conclusion: Such strategies can be used to produce high levels of desired protein in target cells as a therapeutic target. The produced recombinant cells may present a valuable platform to analyze the effect of DPPA2 ectopic expression in target cells. Moreover, the introduction of its potential capacity into the mouse model to evaluate the tumorigenesis of these cancer cells in vivo leads to an understanding of the biological importance of DPPA2 in tumorigenesis. In addition, our purified protein can be used in a mouse model to produce specific antibody developing a reliable detection of DPPA2 existence in any biological fluid through ELISA system
Résumé
Background: Recently, it has been revealed that tyrosine kinase inhibitors [TKIs] act through inducing both oxidative and endoplasmic reticulum [ER] stress in chronic myeloid leukemia cells. However, ER stress signaling triggers both apoptotic and survival processes within cells. Nevertheless, mechanisms by which TKIs avoid the pro-survival effects are not clear. The aim of this study was to evaluate the potential role of oxidative stress in activity of unfolded protein response [UPR] survival pathway within K562 cell line
Methods: The expression of UPR survival target genes, Xbpl, and Grp94 [glucose requiring protein 94] was studied in single and combined exposure to oxidative and ER stress in K562 cell line by quantitative and qualitative PCR
Results: The expression of UPR-related survival gene Grp94 was hampered by exposing to oxidative stress in cell induced with ER stress
Conclusion: Interaction of oxidative and ER stress may role as a mediator influencing UPR signaling activity
Résumé
CatSper genes are a novel family of four sperm-specific calcium channels, which indicate testis-specific expression patterns. Despite the crucial role of CatSper genes in the male reproduction, very little is known about the factors that regulate their expression. The objective of this study was to investigate the effects of vitamin E treatment on the expression of CatSper 1 and CatSper 2 genes as well as sperm quality in the aged male mice. Twenty four 11-12 months old aged male mice and twenty four 2-3-months old young male mice were randomly divided into four groups. Control groups received no injection. The experimental groups of male mice were received intraperitoneal injection of 106 mg/kg vitamin E daily for 35 days. Left testis and cauda epididymides from each mouse were collected on the days 21, 28 and 35 following vitamin E treatment and were used for Real-Time PCR and immunohistochemistry. Also, sperm analysis was performed according to the WHO guidelines given for human sperm examination. Data were analyzed using SPSS software Administration of vitamin E improved sperm parameters in the aged as well as young adult male mice. In addition, the expression of CatSper genes increased following vitamin E treatment. Also, intensity of signal for CatSper1 and CatSper2 increased in the head and middle piece of sperm in experimental group as compared to those of control ones. The vitamin E treatment significantly improved the sperm quality, especially in terms of sperm motility, count and morphology rate. Furthermore, CatSper genes expression could be up-regulated by the vitamin E treatment