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25. Sal-Like Protein 4 Transcription Factor: A Significant Diagnostic Biomarker Involved in Childhood ALL Resistance and Relapse

Author

Ohadi, Farzaneh, Rahgozar, Soheila, and Ghodousi, Elaheh Sadat

Subjects
relapse, SALL4, ABC transporters, multidrug resistance, Cancer Management and Research, childhood acute lymphoblastic leukemia, ABCA3, eye diseases, Original Research
Abstract

Farzaneh Ohadi, Soheila Rahgozar, Elaheh Sadat Ghodousi Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, IranCorrespondence: Soheila RahgozarDepartment of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Hezar Jarib Street, Isfahan 81746-73441, IranTel +98 313 793 2455Fax +98 313 793 2456Email rahgozar@sci.ui.ac.irPurpose: Sal‐like protein 4 transcription factor (SALL4) is a stem cell transcription factor that plays an essential role in the maintenance and self-renewal of embryonic and hematopoietic stem cells, functioning as an oncogene in several cancers. However, the role of SALL4 in the biological behavior of childhood acute lymphoblastic leukemia and its relationship with multidrug resistance and relapse has remained largely unknown.Patients and Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to characterize the expression pattern of SALL4 in the bone marrow samples of 43 patients with Philadelphia negative ALL and 18 children in the non-cancer control group. The presence of minimal residual disease was measured a year after the initial therapy using SSCP (single-strand conformation polymorphism). In addition, the correlation between the expression of SALL4 and ABCA3 in relapsed patients was analyzed statistically.Results: Results showed an overexpression of SALL4 in de novo patients compared with the control group (P=0.0001, AUC= 0.93), indicating the importance of this gene in the induction of leukemia. A significant increase in the ABCA3 expression levels was revealed in the relapsed patients, in comparison with the drug-sensitive group (P = 0.0005). The leukemogenetic effect of SALL4 can be related to the effect of this gene on the maintenance of pluripotency in cancer stem cells. Results also suggest that the expression of SALL4 can be considered as a diagnostic marker for pediatric ALL. Moreover, SALL4 expression levels in the minimal residual disease positive (mrd+) ALL group was significantly higher than those in the mrd− group (p=0.0001, AUC= 0.92).Conclusion: These data demonstrate the prognostic impact of SALL4 in childhood ALL. Our findings also indicated a direct correlation between the mRNA expression levels of SALL4 and ABCA3 transporter in the relapsed group of ALL patients (r=0.7). These results describe a possible mechanism by which SALL4 may lead to the development of multidrug resistance.Keywords: childhood acute lymphoblastic leukemia, multidrug resistance, SALL4, ABC transporters, relapse, ABCA3

Published
2020

26. Evidence of still ongoing convergence evolution of the lactase persistence [T.sub.-13910] alleles in humans

Author

Enattah, Nabil Sabri, Trudeau, Aimee, Pimenoff, Ville, Maiuri, Luigi, Auricchio, Salvatore, Greco, Luigi, Rossi, Mauro, Lentze, Michael, Seo, J.K., Rahgozar, Soheila, Khalil, Insaf, Alifrangis, Michael, Natah, Sirajedin, Groop, Leif, Shaat, Nael, Kozlov, Andrew, Verschubskaya, Galina, Comas, David, Bulayeva, Kazima, Mehdi, S.. Qasim, Terwilliger, Joseph D., Sahi, Timo, Savilahti, Erkki, Perola, Markus, Sajantila, Antti, Jarvela, Irma, and Peltonen, Leena

Subjects
Allelomorphism -- Research, DNA -- Research, Convergence (Biology) -- Research, Biological sciences
Abstract

The background of the alleles carrying the critical variant in 1611 DNA samples from 37 populations were analyzed and concluded that the [T.sub.-13910] allele was independently introduced more than once.

Published
2007

27. Detection and elimination of Proteus vulgaris and Klebsiella pneumoniae in nude mice.

Author

Dorcheh, Somayeh Najafi, Abedi, Marjan, Rahgozar, Soheila, Moshtaghian, Jamal, Talei, Daryush, and Talebi, Ardeshir

Subjects
KLEBSIELLA pneumoniae, MICROBIAL sensitivity tests, PRICE maintenance, ANIMAL rescue, MICE, ANTIBIOTIC prophylaxis
Abstract

Introduction: Nude mice are immunodeficient, commonly used model organisms for studying human diseases. Despite the high purchase price and maintenance cost of these animals, few studies have aimed to eradicate inevitable infectious complications which may arise during in-vivo studies. The current project is a practical guideline for detecting and elimination of possible infections developed in nude mice. Materials and Methods: Followed by transplantation of human leukemic cell lines into C57BL/6 nude mice, body weights were monitored daily. After a sudden death of 7 mice during 4 consecutive days, all beddings were discarded and cages, food and water bottles were disinfected immediately. Automated blood culture analyses were carried out using radiometric BACTEC systems, and specific antibiotics were administered based on applied microbial growth inhibition assays. Results: Proteus vulgaris and Klebsiella pneumoniae were identified, respectively, according to the radiometric assays. According to the antimicrobial susceptibility tests, 132mg/kg ciprofloxacin was administered orally for 2 weeks, followed by 500mg/kg cefixime for one week. Meanwhile, low-dose cefixime (132 mg/kg) was applied as a prophylactic treatment. Automated blood cultures showed elimination of both bacterial infections followed by demand treatments. Low-dose antibiotic prophylaxis saved uninfected mice from developing infections. Discussion and Conclusion: Results showed that precise monitoring of nude mice health conditions help early detection of the possibly grown infections. Ciprofloxacin and cefixime are the antibiotics of choice for eradication of Proteus vulgaris and Klebsiella pneumoniae. On the other hand, low-dose antibiotic prophylactic treatment may rescue these expensive animals from possible infections and provide suitable environment for performing in-vivo experiments. [ABSTRACT FROM AUTHOR]

Published
2022
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32. The antitoxic effects of quercetin and quercetin-conjugated iron oxide nanoparticles (QNPs) against H2O2-induced toxicity in PC12 cells

Author

Yarjanli,Zahra, Ghaedi,Kamran, Esmaeili,Abolghasem, Zarrabi,Ali, and Rahgozar,Soheila

Subjects
International Journal of Nanomedicine
Abstract

Zahra Yarjanli,1 Kamran Ghaedi,1 Abolghasem Esmaeili,1 Ali Zarrabi,2,3 Soheila Rahgozar11Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran; 2Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran; 3Sabanci University Nanotechnology Research and Application Center (SUNUM), Istanbul, TurkeyCorrespondence: Kamran GhaediDepartment of Biology, Faculty of Sciences, University of Isfahan, Hezar Jerib Ave., Azadi Square, 81746 Isfahan, IranTel +98 313 793 2479Fax +98 313 793 2456Email kamranghaedi@sci.ui.ac.irBackground: We recently showed that quercetin-conjugated iron oxide nanoparticles (QNPs) promoted the bioavailability of quercetin (Qu) in the brain of rats and improved the learning and memory of diabetic rats. In this study, we characterized the modifications in the antitoxic effects of Qu after conjugation.Materials and methods: We conjugated Qu to dextran-coated iron oxide nanoparticles (DNPs) and characterized DNPs and QNPs using FTIR, XRD, DLS, Fe-SEM, and EDX analyzes. The antiradical properties of Qu, DNPs, and QNPs were compared by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity assay. Catalase-like activities of DNPs and QNPs were estimated using catalase activity assay kit, and the antitoxic effects of Qu and QNPs were evaluated with spectrophotometry, MTT assay, flow cytometry, and real-time q-PCR.Results: Qu had a stronger anti-radical activity than DNPs and its activity decreased after being conjugated to DNPs. The catalase-like activity of DNPs remained intact after conjugation. DNPs had less toxicity on PC12 cells viabilities as compared to free Qu, and the conjugation of Qu with DNPs attenuated its cytotoxicity. Furthermore, MTT assay results indicated 24h pretreatment with Qu had more protective effects than QNPs against H2O2-induced cytotoxicity, while Qu and QNPs had the same effects for 48 and 72h incubation. Although the total antioxidant capacity of Qu was attenuated after conjugation, the results of flow cytometry and real-time q-PCR confirmed that 24h pretreatment with the low concentrations of Qu and QNPs had the similar antioxidant, anti-inflammatory, and anti-apoptotic effects against the cytotoxicity of H2O2.Conclusion: Qu and QNPs showed the similar protective activities against H2O2-induced toxicity in PC12 cells. Given the fact that QNPs have magnetic properties, they may serve as suitable carriers to be used in neural research and treatment.Keywords: antitoxic effect, iron oxide nanoparticles, quercetin, PC12 cells

Published
2019

33. The Hedgehog signal transducer Smoothened and microRNA-326: pathogenesis and regulation of drug resistance in pediatric B-cell acute lymphoblastic leukemia

Author

Sheybani,Zahra, Rahgozar,Soheila, and Ghodousi,Elaheh Sadat

Subjects
Cancer Management and Research
Abstract

Zahra Sheybani, Soheila Rahgozar, Elaheh Sadat GhodousiDepartment of Biology, Faculty of Science, University of Isfahan, Isfahan, IranPurpose: Multidrug resistance (MDR) and the subsequent disease relapse are the major causes of childhood acute lymphoblastic leukemia (ALL) related death. The Hedgehog (Hh) signaling pathway can contribute to cancer MDR. In the current study, Smoothened (Smo) was selected as the experimental target due to its importance in the Hh pathway in order to evaluate its probable role in pediatric B-ALL drug resistance.Patients and methods: The study included 27 pediatric B-ALL and 16 control bone marrow samples. Quantitative RT-PCR was used to investigate the expression levels of Smo and miR-326 as the key players of the Hh pathway. Western blot analysis was performed. The presence of minimal residual disease was studied using PCR-SSCP. The association between Smo expression and drug resistance was analyzed statistically.Results: Results showed a significant increase in the Smo expression levels in drug-resistant patients in comparison with drug-sensitive children with B-ALL (P=0.0128, AUC=0.82). A considerable negative association between miR-326 and Smo expression levels was identified (r=−0.624, P=0.002). A binomial test confirmed the regulatory role of miR-326 on the translational repression of Smo (P=0.031). Statistics showed no association between Smo and ABCA2 expression levels. However, a significant positive correlation was observed between the Smo and ABCA3 transcripts in the resistant ALL children (r=0.607, P=0.016).Conclusion: Data revealed the possible oncogenic impact of Smo on leukemogenesis and drug resistance in pediatric B-ALL. Upregulation of Smo was introduced, for the first time, as a prognostic factor for drug resistance in childhood B-ALL. To the best of our knowledge, this is the first study that shows a positive correlation between Smo and ABCA3 expression levels in pediatric B-ALL, explaining a possible mechanism for the development of drug resistance in this cancer. Moreover, the current project revealed a negative modulatory effect of miR-326 on the expression levels of Smo.Keywords: Hh pathway, Smo, miR-326, ABC transporter

Published
2019

35. 6-Shogaol induces apoptosis in acute lymphoblastic leukaemia cells by targeting p53 signalling pathway and generation of reactive oxygen species.

Author

Dorcheh, Somayeh Najafi, Rahgozar, Soheila, and Talei, Daryush

Subjects
LYMPHOBLASTIC leukemia, ACUTE leukemia, REACTIVE oxygen species, CHEMOTHERAPY complications, SURVIVAL rate, APOPTOSIS
Abstract

Combination therapies, using medicinal herbs, are broadly recommended to attenuate the chemotherapy adverse effects. Based on our previous findings considering the anti-leukaemic effects of ginger extract on acute lymphoblastic leukaemia (ALL) cells, the present study was aimed to investigate the anti-cancer role of this pharmaceutical plant on ALL mice models. Moreover, we worked towards identifying the most anti-leukaemic derivative of ginger and the mechanism through which it may exert its cytotoxic impact. In vivo experiments were performed using five groups of six C57BL/6 nude mice, and the anti-leukaemic activity of ginger extract alone or in combination with methotrexate (MTX) was examined. Results showed increased survival rate and reduced damages in mice brain and liver tissues. Subsequently, MTT assay demonstrated synergistic growth inhibitory effect of 6-shogaol (6Sh) and MTX on ALL cell lines and patients primary cells. Eventually, the molecular anti-neoplastic mechanism of 6Sh was evaluated using Bioinformatics. Flow cytometry illustrated 6Sh-mediated apoptosis in Nalm-6 cells confirmed by Western blotting and RT-PCR assays. Further analyses exhibited the generation of reactive oxygen species (ROS) through 6Sh. The current study revealed the in vivo novel anti-leukaemic role of ginger extract, promoted by MTX. Moreover, 6-shogaol was introduced as the major player of ginger cytotoxicity through inducing p53 activity and ROS generation. [ABSTRACT FROM AUTHOR]

Published
2021
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40. Quercetin-Conjugated Superparamagnetic Iron Oxide Nanoparticles Protect AlCl3-Induced Neurotoxicity in a Rat Model of Alzheimer's Disease via Antioxidant Genes, APP Gene, and miRNA-101.

Author

Amanzadeh Jajin, Elnaz, Esmaeili, Abolghasem, Rahgozar, Soheila, and Noorbakhshnia, Maryam

Subjects
IRON oxide nanoparticles, ANIMAL disease models, ALZHEIMER'S disease, QUERCETIN, ANTIOXIDANTS, LABORATORY rats, IMMOBILIZATION stress
Abstract

Alzheimer's disease (AD) is a neurodegenerative disease with cognitive impairment. Oxidative stress in neurons is considered as a reason for development of AD. Antioxidant agents such as quercetin slow down AD progression, but the usage of this flavonoid has limitations because of its low bioavailability. We hypothesized that quercetin-conjugated superparamagnetic iron oxide nanoparticles (QT-SPIONs) have a better neuroprotective effect on AD than free quercetin and regulates the antioxidant, apoptotic, and APP gene, and miRNA-101. In this study, male Wistar rats were subjected to AlCl3, AlCl3 + QT, AlCl3 + SPION, and AlCl3 + QT-SPION for 42 consecutive days. Behavioral tests and qPCR were used to evaluate the efficiency of treatments. Results of behavioral tests revealed that the intensity of cognitive impairment was decelerated at both the middle and end of the treatment period. The effect of QT-SPIONs on learning and memory deficits were closely similar to the control group. The increase in expression levels of APP gene and the decrease in mir101 led to the development of AD symptoms in rats treated with AlCl3 while these results were reversed in the AlCl3 + QT-SPIONs group. This group showed similar results with the control group. QT-SPION also decreased the expression levels of antioxidant enzymes along with increases in expression levels of anti-apoptotic genes. Accordingly, the antioxidant effect of QT-SPION inhibited progression of cognitive impairment via sustaining the balance of antioxidant enzymes in the hippocampus of AD model rats. [ABSTRACT FROM AUTHOR]

Published
2021
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42. The translational expression of ABCA2 and ABCA3 is a strong prognostic biomarker for multidrug resistance in pediatric acute lymphoblastic leukemia

Author

Aberuyi,Narges, Rahgozar,Soheila, Khosravi Dehaghi,Zohreh, Moafi,Alireza, Masotti,Andrea, Paolini,Alessandro, Aberuyi,Narges, Rahgozar,Soheila, Khosravi Dehaghi,Zohreh, Moafi,Alireza, Masotti,Andrea, and Paolini,Alessandro

Abstract

Narges Aberuyi,1 Soheila Rahgozar,1 Zohreh Khosravi Dehaghi,1 Alireza Moafi,2 Andrea Masotti,3,* Alessandro Paolini3,* 1Department of Biology, Faculty of Science, University of Isfahan, 2Department of Pediatric-Hematology-Oncology, Sayed-ol-Shohada Hospital, Isfahan University of Medical Sciences, Isfahan, Iran; 3Gene Expression – Microarrays Laboratory, Bambino Gesù Children’s Hospital-Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Rome, Italy *These authors contributed equally to the manuscript Purpose: The aim of this work was to study the correlation between the expressions of the ABCA2 and ABCA3 genes at the mRNA and protein levels in children with acute lymphoblastic leukemia (ALL) and the effects of this association on multidrug resistance (MDR).Materials and methods: Sixty-nine children with de novo ALL and 25 controls were enrolled in the study. Mononuclear cells were isolated from the bone marrow. The mRNA levels of ABCA2 and ABCA3 were measured by real-time polymerase chain reaction (PCR). Samples with high mRNA levels were assessed for respective protein levels by Western blotting. Following the first year of treatment, persistent monoclonality of T-cell gamma receptors or immunoglobulin H (IgH) gene rearrangement was assessed and considered as the MDR. The tertiary structure of ABCA2 was predicted using Phyre2 and I-TASSER web systems and compared to that of ABCA3, which has been previously reported. Molecular docking was performed using DOCK 6.7.Results: Real-time quantitative PCR (qRT-PCR) showed high levels of ABCA2 and ABCA3 mRNAs in 13 and 17 samples, respectively. Among them, five and eight individuals demonstrated high levels of ABCA2 and ABCA3, respectively. Response to chemotherapy was significantly decreased (P=0.001) when the mRNA and protein of both genes were overexpressed compared to individuals with high transcriptional levels of either ABCA2 or ABCA3 alone. Close similarity between AB

Published
2017

46. Recovery of MicroRNA from Stored Bone Marrow Aspirate Slides.

Author

Ghodousi, Elaheh Sadat and Rahgozar, Soheila

Subjects
*LEUKEMIA diagnosis, *COLLECTION & preservation of biological specimens, *BONE marrow, *CHEMICAL reagents, *CONVALESCENCE, *DNA, *FROZEN tissue sections, *GENE expression, *POLYMERASE chain reaction, *PRESERVATION of organs, tissues, etc., *REVERSE transcriptase polymerase chain reaction, *DATA analysis software, *MICRORNA, *CHILDREN
Abstract

Background: Archived bone marrow aspirate slides are almost infinite, readily available resource of biospecimens that enable retrospective molecular investigations of diseases. RNAs obtained from slides has limitations in utility because of their low quality and highly fragmented nature. MicroRNAs are small (<22 nt) noncoding RNAs with various cellular regulatory roles. Due to their small size, microRNAs are less prone to degradation and modification, therefore, can be preserved well in archived tissues. Methods: The current study investigated the efficacy of archived bone marrow aspirate slides for miRNA expression analysis in pediatric leukemia. Total RNA was isolated from air-dried unstained archived slides using High pure miRNA isolation Kit with some modifications and from fresh samples using TRizol. After cDNA synthesis, RT-qPCR was then carried out using specific hsa-miR-326 LNA primers. Finally, statistical analyses were conducted using GraphPad Prism 6 software. Results: The difference observed in miRNA expression due to disease state was far greater than the differences between archived slides and their matching fresh bone marrow specimens. In fact, the expression of archival slide smears for the miR-326 closely mimicked that of fresh-frozen tissues (0.035±0.04 vs. 0.03±0.04) (Mean±SD, p>0.05). Differential expression of hsa-miR-326 was detected between leukemic and non-leukemic samples from archived slides or fresh frozen bone marrows. Conclusion: The demonstration that archived bone marrow aspirate slides can be utilized for miRNA expression studies offers tremendous potential for future investigations into the role that miRNAs play in the development and long term outcome of hematologic, as well as non-hematologic diseases. [ABSTRACT FROM AUTHOR]

Published
2019

47. Integrative computational in-depth analysis of dysregulated miRNA-mRNA interactions in drug-resistant pediatric acute lymphoblastic leukemia cells: an attempt to obtain new potential gene-miRNA pathways involved in response to treatment

Author

Mesrian Tanha, Hamzeh, Mojtabavi Naeini, Marjan, Rahgozar, Soheila, Moafi, Alireza, Honardoost, Mohamad Amin, Mesrian Tanha, Hamzeh, Mojtabavi Naeini, Marjan, Rahgozar, Soheila, Moafi, Alireza, and Honardoost, Mohamad Amin

Abstract

Acute lymphoblastic leukemia (ALL) is the major neoplasia type among children. Despite the tremendous success of current treatment strategies, drug resistance still remains a major cause of chemotherapy failure and relapse in pediatric patients. Overwhelming evidence illustrates that microRNAs (miRNAs) act as post-transcriptional regulators of drug-resistance-related genes. The current study was aimed at how dysregulated miRNA-mRNA-signaling pathway interaction networks mediate resistance to four commonly used chemotherapy agents in pediatric ALL, including asparaginase, daunorubicin, prednisolone, and vincristine. Using public expression microarray datasets, a holistic in silico approach was utilized to investigate candidate drug resistance miRNA-mRNA-signaling pathway interaction networks in pediatric ALL. Our systems biology approach nominated significant drug resistance and cross-resistance miRNAs, mRNAs, and cell signaling pathways based on anti-correlative relationship between miRNA and mRNA expression pattern. To sum up, our systemic analysis disclosed either a new potential role of miRNAs, or a possible mechanism of cellular drug resistance, in chemotherapy resistance of pediatric ALL. The current study may shed light on predicting drug response and overcoming drug resistance in childhood ALL for subsequent generations of chemotherapies.

Published
2016

50. Modified tetra-primer ARMS PCR as a single-nucleotide polymorphism genotyping tool

Author

Mesrian Tanha, Hamzeh, Mojtabavi Naeini, Marjan, Rahgozar, Soheila, Mahdi Rasa, Seyed Mohammad, Vallian, Sadeq, Mesrian Tanha, Hamzeh, Mojtabavi Naeini, Marjan, Rahgozar, Soheila, Mahdi Rasa, Seyed Mohammad, and Vallian, Sadeq

Abstract

Objectives: Genotyping of single-nucleotide polymorphisms (SNPs) has been applied in various genetic contexts. Tetra-primer amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) is reported as a prominent assay for SNP genotyping. However, there were published data that may question the reliability of this method on some occasions, in addition to a laborious and time-consuming procedure of the optimization step. In the current study, a new SNP genotyping method named modified tetra-primer ARMS (MTPA) PCR was developed based on tetra-primer ARMS PCR. Design and Methods: The modified method has two improvements in its instruction, including equalization of outer primer and inner primer strength by additional mismatch in outer primers, and consideration of equal annealing temperature of specific fragments more than melting temperature of primers. Advantageously, a new computer software was provided for designing primers based on novel concepts. Results: The usual tetra-primer ARMS PCR has a laborious process for optimization. In nonoptimal PCR programs, identification of the accurate genotype was found to be very difficult. However, in MTPA PCR, equalization of the amplicons and primer strength leads to increasing specificity and convenience of genotyping, which was validated by sequencing. Conclusions: In the MTPA PCR technique, a new mismatch at −2 positions of outer primers and equal annealing temperature improve the genotyping procedure. Together, the introduced method could be suggested as a powerful tool for genotyping single-nucleotide mutations and polymorphisms.

Published
2015

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