Cell Journal (Yakhteh)

583-592

Authors: Najmaldin Saki, Saeid Abroun, Masoud Soleimani, Maria Kavianpour, Mohammad Shahjahani, Javad Mohammadi-Asl, Saeideh Hajizamani

Today the regulatory role of microRNAs (miRs) is well characterized in many diverse cellular processes. MiR-based regulation is categorized under epigenetic regulatory mechanisms. These small non-coding RNAs participate in producing and maturing erythrocytes, expressing hematopoietic factors and regulating expression of globin genes by post-transcriptional gene silencing. The changes in expression of miRs (miR-144/-320/-451/-503) in thalassemic/sickle cells compared with normal erythrocytes may cause clinical severity. According to the suppressive effects of certain miRs (miR-15a/-16-1/-23a/-26b/-27a/-451) on a number of transcription factors [myeloblastosis oncogene (MYB), B-cell lymphoma 11A (BCL11A), GATA1, Krüppel-like factor 3 (KLF3) and specificity protein 1 (Sp1)] during β globin gene expression, It has been possible to increasing γ globin gene expression and fetal hemoglobin (HbF) production. Therefore, this strategy can be used as a novel therapy in infusing HbF and improving clinical complications of patients with hemoglobinopathies.

601-607

Authors: Elahe Motevaseli, Rosa Azam, Seyed Mohammad Akrami, Mohammadali Mazlomy, Mojtaba Saffari,Mohammad Hossein Modarressi, Maryam Daneshvar, Soudeh Ghafouri-Fard,

Objective: The aim of this study was to clarify the mechanism by which lactobacilli exert their cytotoxic effects on cervical cancer cells. In addition, we aimed to evaluate the effect of lactobacilli on the expression of human papilloma virus (HPV) oncogenes. Materials and Methods: In this experimental study, using quantitative real-time polymerase chain reaction (PCR), we analyzed the expression of CASP3 and three autophagy genes [ATG14, BECN1 and alpha 2 catalytic subunit of AMPK (PRKAA2)] along with HPV18 E6 and E7 genes in HeLa cells before and after treatment with Lactobacillus crispatus and Lactobacillus rhamnosus culture supernatants. Results: The expression of CASP3 and autophagy genes in HeLa cells was decreased after treatment with lactobacilli culture supernatants. However, this decrease was not significant for PRKAA2 when compared with controls. In addition, expression of HPV E6 was significantly decreased after treatment with lactobacilli culture supernatants. Conclusion: Lactobacilli culture supernatants can decrease expression of ATG14 and BECN1 as well as the HPV E6 oncogene. It has been demonstrated that the main changes occurring during cervical carcinogenesis in cell machinery can be reversed by suppression of HPV oncogenes. Therefore, downregulation of HPV E6 by lactobacilli may have therapeutic potential for cervical cancer. As the role of autophagy in cancer is complicated, further work is required to clarify the link between downregulation of autophagy genes and antiproliferative effects exerted by lactobacilli. Keywords: HPV, Lactobacillus, Autophagy

617-628

Authors: Monireh Hajimoradi, Zuhair Mohammad Hassan, Marzieh Ebrahimi, Masoud Soleimani,Mahdieh Bakhshi, Javad Firouzi, Fazel Sahraneshin Samani

Objective: Gastric cancer (GC) is widely associated with chronic inflammation. The pro inflammatory microenvironment provides conditions that disrupt stem/progenitor cell proliferation and differentiation. The signal transducer and activator of transcription- 3 (STAT3) signaling pathway is involved in inflammation and also contributes to the maintenance of embryonic stem cell (ESCs) pluripotency. Here, we have investigated the activation status of STAT3 in GC stem-like cells (GCSLCs). Materials and Methods: In this experimental research, CSLCs derived from the human GC cell line MKN-45 and patient specimens, through spheroid body formation, characterized and then assayed for the STAT3 transcription factor expression in mRNA and protein level further to its activation. Results: Spheroid cells showed higher potential for spheroid formation than the parental cells. Furthemore, stemness genes NANOG, c-MYC and SOX-2 were over expressed in spheroids of MKN-45 and in patient samples. In MKN-45 spheroid cells, epithelial mesenchymal transition (EMT) related markers CDH2, SNAIL2, TWIST and VIMENTIN were upregulated (P<0.05), but we observed no change in expression of the E-cadherin epithelial marker. These cells exhibited more resistance to docetaxel (DTX) when compared with parental cells (P<0.05) according to the MTS assay. Although immunostaining and Western blotting showed expression of the STAT3 protein in both spheroids and parents, the mRNA level of STAT3 in spheroids was higher than the parents. Nuclear translocation of STAT3 was accompanied by more intensive phospho-STAT3 (p-STAT3) in spheroid structures relative to the parent cells according to flow cytometry analysis (P<0.05). Conclusion: The present findings point to STAT3 over activation in GCSLCs. Complementary experiments are required to extend the role of STAT3 in stemness features and invasion properties of GCSCs and to consider the STAT3 pathway for CSC targeted therapy. Keywords: Gastric Cancer, Cancer Stem Cells, Spheroid, STAT3, EMT

639-647

Authors: Zoleikha Golipoor, Fereshteh Mehraein, Fariba Zafari,Akram Alizadeh,Shima Ababzadeh,Maryam Baazm

Objective: Bone marrow (BM) is one of the major hematopoietic organs in postnatal life that consists of a heterogeneous population of stem cells which have been previously described. Recently, a rare population of stem cells that are called very small embryonic-like (VSEL) stem cells has been found in the BM. These cells express several developmental markers of pluripotent stem cells and can be mobilized into peripheral blood (PB) in response to tissue injury. In this study we have attempted to investigate the ability of these cells to migrate toward an injured spinal cord after transplantation through the tail vein in a rat model. Materials and Methods: In this experimental study, VSELs were isolated from total BM cells using a fluorescent activated cell sorting (FACS) system and sca1 and stage specific embryonic antigen (SSEA-1) antibodies. After isolation, VSELs were cultured for 7 days on C2C12 as the feeder layer. Then, VSELs were labeled with 1,1´-dioctadecyl-3,3,3´,3´- tetramethylindocarbocyanine perchlorate (DiI) and transplanted into the rat spinal cord injury (SCI) model via the tail vein. Finally, we sought to determine the presence of VSELs in the lesion site. Results: We isolated a high number of VSELs from the BM. After cultivation, the VSELs colonies were positive for SSEA-1, Oct4 and Sca1. At one month after transplantation, real-time polymerase chain reaction analysis confirmed a significantly increased expression level of Oct4 and SSEA-1 positive cells at the injury site. Conclusion: VSELs have the capability to migrate and localize in an injured spinal cord after transplantation. Keywords: Homing, Spinal Cord Injury, Migration

659-667

Authors: Marzieh Moghadas,Mohammad Amin Edalatmanesh,Reza Robati

Objective: The present study investigated the effects of gallic acid (GA) administration on trimethyltin chloride (TMT) induced anxiety, depression, and hippocampal neurodegeneration in rats. Materials and Methods: In this experimental study, the rats received intraperitoneal (i.p.) injections of TMT (8 mg/kg). The animals received either GA (50, 100 and 150 mg/kg) or saline as the vehicle for 14 consecutive days. We measured depression and anxiety levels of the rats by conducting the behavioral tail suspension (TST), elevatedplusmaze (EPM), and novelty suppressed feeding (NSF) tests. Histological analyses were then used to determine the cell densities of different hippocampal subdivisions. The data were analyzed with ANOVA and Tukey’s post hoc test. Results: GA administration ameliorated anxiety and depression in the behavioral tests. The cell densities in the CA1, CA2, CA3 and DG hippocampal subdivisionsfrom GA-treated rats were higher than saline treated rats. Conclusion: GA treatment against TMT-induced hippocampal degeneration altered cellular loss in the hippocampus and ameliorated the depression-anxiety state in rats. Keywords: Trimethyltin, Gallic Acid, Hippocampus, Cell Density

678-691

Authors: Zahra Zandieh,Fatemehsadat Amjadi, Mahnaz Ashrafi, Abbas Aflatoonian, Alireza Fazeli, Reza Aflatoonian

Objective: Toll like receptors (TLRs) are one of the main components of the innate immune system. It has been reported that expression of these receptors are altered in the female reproductive tract (FRT) during menstrual cycle. Here we used a fallopian tube epithelial cell line (OE-E6/E7) to evaluate the effect of two sex hormones in modulating TLR expression. Materials and Methods: In this experimental study, initially TLR gene expression in OEE6/ E7 cells was evaluated and compared with that of fallopian tube tissue using quantitative real time-polymerase chain reaction (qRT-PCR) and immunostaining. Thereafter, OE-E6/E7 cells were cultured with different concentrations of estradiol and progesterone, and combination of both. qRT-PCR was performed to reveal any changes in expression of TLR genes as a result of hormonal treatment. Results: TLR1-10 genes were expressed in human fallopian tube tissue. TLR1-6 genes and their respective proteins were expressed in the OE-E6/E7 cell line. Although estradiol and progesterone separately had no significant effect on TLR expression, their combined treatment altered the expression of TLRs in this cell line. Also, the pattern of TLR expression in preovulation (P), mensturation (M) and window of implantation (W) were the same for all TLRs with no significant differences between P, M and W groups. Conclusion: These data show the significant involvement of the combination of estradiol and progesterone in modulation of TLR gene expression in this human fallopian tube cell line. Further experiments may reveal the regulatory mechanism and signalling pathway behind the effect of sex hormones in modulating TLRs in the human FRT. Keywords: Estradiol, Progesterone, Fallopian Tube, Toll Like Receptors

701-710

Authors: Morteza Abouzaripour,Parichehr Pasbakhsh,Nader Atlasi,Abdol Hossein Shahverdi,Reza Mahmoudi,Iraj Ragerdi Kashani

Objective: Bone marrow has recently been recognized as a novel source of stem cells for the treatment of wide range of diseases. A number of studies on murine bone marrow have shown a homogenous population of rare stage-specific embryonic antigen 1 (SSEA-1) positive cells that express markers of pluripotent stem cells. This study focuses on SSEA-1 positive cells isolated from murine bone marrow in an attempt to differentiate them into insulin-secreting cells (ISCs) in order to investigate their differentiation potential for future use in cell therapy. Materials and Methods: This study is an experimental research. Mouse SSEA-1 positive cells were isolated by Magnetic-activated cell sorting (MACS) followed by characterization with flow cytometry. Induced SSEA-1 positive cells were differentiated into ISCs with specific differentiation media. In order to evaluate differentiation quality and analysis, dithizone (DTZ) staining was use, followed by reverse transcription polymerase chain reaction (RT-PCR), immunocytochemistry and insulin secretion assay. Statistical results were analyzed by one-way ANOVA. Results: The results achieved in this study reveal that mouse bone marrow contains a population of SSEA-1 positive cells that expresses pluripotent stem cells markers such as SSEA-1, octamer-binding transcription factor 4 (OCT-4) detected by immunocytochemistry and C-X-C chemokine receptor type 4 (CXCR4) and stem cell antigen-1 (SCA-1) detected by flow cytometric analysis. SSEA-1 positive cells can differentiate into ISCs cell clusters as evidenced by their DTZ positive staining and expression of genes such as Pdx1 (pancreatic transcription factors), Ngn3 (endocrine progenitor marker), Insulin1 and Insulin2 (pancreaticβ-cell markers). Additionally, our results demonstrate expression of PDX1 and GLUT2 protein and insulin secretion in response to a glucose challenge in the differentiated cells. Conclusion: Our study clearly demonstrates the potential of SSEA-1 positive cells to differentiate into insulin secreting cells in defined culture conditions for clinical applications. Keywords: Stage-Specific Embryonic Antigen, Insulin-Secreting Cells, Cell Differentiation, Diabetes Mellitus

720-729

Authors: Layasadat Khorsandi, Sadegh Saremy, Ali Khodadadi, Fereshteh Dehbashi

Objective: To evaluate the effect of Exendine-4 (EX-4), a Glucagon-like peptide 1 (GLP-1) receptor agonist, on the differentiation of insulin-secreting cells (IPCs) from rat adipose-derived mesenchymal stem cells(ADMSCs). Materials and Methods: In this experimental study, ADMSCs were isolated from rat adipose tissue and exposed to induction media with or without EX-4. After induction, the existence of IPCs was confirmed by morphology analysis, expression pattern analysis of islet-specific genes (Pdx-1, Glut-2 and Insulin) and insulin synthesis and secretion. Results: IPCs induced in presence of EX-4 were morphologically similar to pancreatic islet-like cells. Expression of Pdx-1, Glut-2 and Insulin genes in EX-4 treated cells was significantly higher than the cells exposed to differentiation media without EX-4. Compared to EX-4 untreated ADMSCs, insulin release from EX-4 treated ADMSCs showed a nearly 2.5 fold (P<0.05) increase when exposed to a high glucose (25 mM) medium. The percentage of insulin positive cells in the EX-4 treated group was approximately 4-fold higher than in the EX-4 untreated ADMSCs. Conclusion: The present study has demonstrated that EX-4 enhances the differentiation of ADMSCs into IPCs. Improvement of this method may help the formation of an unlimited source of cells for transplantation. Keywords: Exendin-4, Differentiation, Insulin-Secreting Cells, Adipose Tissue, Mesenchymal Stem Cells

740-747

Authors: Maryam Sadat Fakhri-Bafghi, Seyedeh Farnaz Ghasemi-Niri, Sara Mostafalou, Mona Navaei-Nigjeh,Maryam Baeeri, Azadeh Mohammadirad, Mohammad Abdollahi

Objective: Organophosphorus (OP) compounds are used to control pests, however they can reach the food chain and enter the human body causing serious health problems by means of acetylcholinesterase (AChE) inhibition and oxidative stress (OS). Among the OPs, chlorpyrifos (CHP), malathion (MAL), and diazinon (DIA) are commonly used for commercial extermination purposes, in addition to veterinary practices, domestic, agriculture and public health applications. Two new recently registered medicines that contain selenium and other antioxidants, IMOD and angipars (ANG), have shown beneficial effects for OS related disorders. This study examines the effect of selenium-based medicines on toxicity of three common OP compounds in erythrocytes. Materials and Methods: In the present experimental study, we determined the efficacy of IMOD and ANG on OS induced by three mentioned OP pesticides in human erythrocytes in vitro. After dose-response studies, AChE, lipid peroxidation (LPO), total antioxidant power (TAP) and total thiol molecules (TTM) were measured in erythrocytes after exposure to OPs alone and in combined treatment with IMOD or ANG. Results: AChE activity, TAP and TTM reduced in erythrocytes exposed to CHP, MAL and DIA while they were restored in the presence of ANG and IMOD. ANG and IMOD reduced the OPs-induced elevation of LPO. Conclusion: The present study shows the positive effects of IMOD and ANG in reduction of OS and restoration of AChE inhibition induced by CHP, MAL and DIA in erythrocytes in vitro. Keywords: Human Erythrocyte, Organophosphorus, Oxidative Stress

755-755

Authors: Seyed Mohammad Javad Mortazavi, Hossein Mozdarani

We read with great interest an article by Shokri et al. entitled "Effects of Wi-Fi (2.45 GH z) exposure on apoptosis, sperm parameters and testicular histomorphometry in rats: a time course study" that is published in the latest issue of the Cell Journal (Vol.17, 2015: 322-331). In this article, Shokri et al. have presented their findings obtained in an experiment on an animal model. These researchers exposed rats to the 2.45 GHz radiation in a chamber with two Wi-Fi antennas on opposite walls of a box. The exposed animals in this study showed a decrease in sperm parameters. We have previously shown that exposure to electromagnetic fields generated by Wi-Fi routers or mobile phone jammers can adversely affect the sperm quality (1-3). The paper published by Shokri et al. is seriously flawed. The first major shortcoming of this paper is its exposure geometry. The authors stated that their exposure system was "a chamber (180 cm×80 cm×70 cm), designed for whole-body exposure of free-moving rats to a Wi-Fi signal. Two Wi-Fi antennas (NanoStation Loco M2, 2.45 GHz, 8.5 dBi, Ubiquiti Networks, Inc. USA) were placed at the center of two sides of the chamber". It should be noted that in this case, the power density can be calculated using the below equation: S=P•G/4πR2 Where S=Power density P=Power input to antenna G=Antenna gain In this light, the geometry used in the study of Shokri et al. makes a very inhomogeneous distribution of power densities. The second shortcoming comes from this point that the authors claimed that their study was performed on a basis that could not affect the hormonal balance "A previous study applied a restrainer to fix space between antenna and rat. Since it was a stressful condition that could probably affect hormonal balance of animals, we tried to assess the effect of radiation on the free moving animals". However, these authors only had a control group and did not use a sham-exposed group to control the animals’s stress and its subsequent hormonal changes. Furthermore, another shortcoming comes from this point that "NanoStation Loco M2" is not a standard Wi-Fi router. As manufacturer reports this device is a compact outdoor communication unit that can be used for devices such as cameras "NanoStation Loco M2 is a compact outdoor unit which includes 2×8 dBi antenna (MIMO) for the 2.4 GHz band”. Therefore, it is misleading to claim that in this study the effects of Wi-Fi exposure on apoptosis are investigated and the title of this paper is indeed incorrect "Effects of Wi-Fi (2.45 GH z) exposure on apoptosis, sperm parameters and testicular histomorphometry in rats”. We hope that these comments are helpful to make more reliable results in the future.