فهرست مطالب

Human Genetics and Genomics - Volume:7 Issue: 1, Jun 2023

Journal of Human Genetics and Genomics
Volume:7 Issue: 1, Jun 2023

  • تاریخ انتشار: 1402/12/26
  • تعداد عناوین: 6
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  • Siroos Tarighi, Maryam Montazeri*, Fatemeh Rohollah Page 1
    Background

    Breast cancer is one of the common malignancies in women, for which doxorubicin (DOX) is widely used in its chemotherapy. Recently, it has been found that DOX affects the expression profile of oncogene genes and miRNAs. In this study, the impacts of DOX on the expressions of the STAT3 gene, miR-874-3p, and miR-337-3p were studied in the MCF-7 breast cancer cell line.

    Methods

    After exposure of MCF-7 cells with DOX, the MTT method was applied for evaluating the cell viability. Apoptosis and necrosis percentages were measured using flow cytometry. Also, the levels of ROS and NF-κB were measured in DOX-treated cells. Then, exosomes secreted from these cells were prepared. The shape of exosomes was studied by SEM. Finally, the expression of bax, bcl-2, p53, casp3, STAT3 gene, and miR-874-3p and miR-337-3p in MCF-7 cells as well as exosomes were evaluated using the RT-PCR technique. Data analysis was done by T-test in GraphPad Prism8 software.

    Results

    The exposure of MCF-7 cells to doxorubicin led to a concentration-dependent decrease in cell viability and increases in apoptosis and necrosis. ROS and NF-κB activity were increased in DOX-treated cells. In DOX-treated cells, decreased expressions of bcl-2 and STAT3 genes and overexpression of bax, p53, casp3, miR-874-3p, and miR-337-3p were observed compared to untreated control cells.

    Conclusion

    One of the mechanisms of the anti-breast cancer effects of DOX is the induction of changes in the expression of oncogenic genes, mediating by downregulating of STAT3 gene and overexpressing miR-874-3p and miR-337-3p. More studies are needed in this field.

    Keywords: Breast cancer, Gene, miRNA, Doxorubicin
  • Tina Tavakoli, Zainab Kasraee, Nooshin Barikrow* Page 2
    Background

    Alzheimer's disease (AD) is a neurodegenerative disease with insidious onset and progressive destruction of behavioral and cognitive functions. Various therapies have been tested to improve or at least effectively change the course of AD. In recent years; stem cell therapy has emerged as a hopeful potential treatment for Alzheimer's. Stem cells can differentiate into various types of cells, including brain cells, potentially replacing damaged cells and improving cognitive function.

    Objectives

    In the present study we investigated the inhibitory effect of mesenchymal stem cells isolated from amniotic membrane (AM-MSCs) on neural progenitor cells (NPC) treated with Scopolamine.

    Methods

    NPC cell was provided by the Iranian Biological Resource Center. To expose these cells to Alzheimer's situations, scopolamine (0.05 mg/ml) treatment has been used. The inhibitory effect of mesenchymal stem cells isolated from the amniotic membrane was evaluated by using the co-cultivation method. The expression of amyloid beta (Aβ), TERM2, Tau, and ABCA7 genes, was assessed in NPC cells co-cultivated with AM-MSC by Real-time PCR. After the Co-culture of AM-MSC and NPC cells for 72 hours, we evaluated the expression of BDNF and CHAT protein in Co-cultured NPC cells by immunocytochemical test.

    Results

    Results of Real-time PCR and Immunocytochemistry showed that in Co-culture of AM-MSC with NPC decreased Aß, TREM2, and Tau gene expression and increased ABCA7 expression. As well, the expression of BDNF and CHAT protein enhanced.

    Conclusions

    AM-MSCs have attracted much consideration. MSCs have the capability of immune regulation, regeneration, and neuroprotection These cells are a potential candidate for cell therapy due to their easy accessibility and compliance with ethical issues.

    Keywords: Mesenchymal stem cells, neural progenitor cell, TERM2, Tau, Aβ, ABCA7
  • Mansoureh Ajami, Omid Moeini, Amir Atashi, Masoud Soleimani, Hossein Dehghani, Monireh Ajami* Page 3
    Background

    Beta-thalassemia is a group of hereditary blood disorders caused by mutations in the β-globin gene cluster resulting in variable phenotypes ranging from severe anemia to clinically asymptomatic individuals. This study aimed to produce an in vitro model of β-thalassemia using CRISPR/Cas9 as an easily programmable, fast, more powerful, and efficient technique.

    Materials and Methods

    Guide RNA (gRNA)-Cas9 co-expression vectors were used for embryonic stem (ES) cell nucleofection. PCR, T7EI, and Hbb-b1 gene sequencing tests were done on extracted DNA to evaluate gene mutation. Following erythroid differentiation of ES cells, analysis of hemoglobin genes and erythroid transcription factors were assessed using a quantitative reverse transcription-polymerase chain reaction.

    Results

    Sequencing data associated with clone 31 confirmed the deletion of 851 nucleotides between exon 2 and 3 in an Hbb-b1 allele in this clone and Indel mutation in exon 2 (-40bp/+38bp) from another allele of Hbb-b1. Significant expression of erythroid transcription factors was observed in wild type, Hbb-b1+/- and Hbb-b1-/- groups. The hbb-b1 gene expression in the Hbb-b1+/- group significantly decreased, although the Hbb-b1-/- group had zero expression.

    Conclusion

    Utilizing an efficient erythroid differentiation method on the CRISPR/Cas9-mediated Hbb-b1 knock-out in ES cells provides accessibility to the laboratory thalassemia model. This method could be used to produce a mouse model of β-thalassemia intermedia (Hbbth1/th1 mice), which are required for the identification of the molecular basis of β-thalassemia and enable testing of the therapeutic approaches such as the recovery of functional β or γ hemoglobin chain.

    Keywords: Beta‐thalassemia, CRISPR‐Cas9 system, Hbb‐b1, Mouse embryonic stem cell
  • Rana Khosravi, Behdokht Jamali*, Morvarid Heidari Page 4
    Background

    Breast cancer is one of the most common malignancies in women for which no suitable treatment has been found yet. Therefore, the present study studied the cytotoxicity effects of tyrosol (TRY) on the Michigan Cancer Foundation-7 (MCF7) breast cancer cell line and L929 normal cells.

    Methods

    MCF7 and L929 cells were cultured red in DMEM-F12 culture medium after preparation and then exposed to 0, 100, 200, and 300 μM of TRY for 72 hours. Cell viability was evaluated using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay, and apoptotic and necrotic cell percentages were determined by flow cytometry. After designing specific primers, the expression levels of bax, p53, and bcl-2 genes were evaluated by RT-PCR. GraphPad Prism software was used for analyzing the data.

    Results

    TRY-treated MCF7 cells showed significantly decreased cell viability in a dose-depended manner. Also, the cell treated with high concentrations of TRY (200 and 300 µM) had a high rate of apoptosis and necrosis (P<0.0001). Reactive oxygen species (ROS) content increased in TRY-treated MCF7 cells. Moreover, the overexpression of bax and p53 and downregulation of bcl-2 was seen in TRY-treated MCF7 cells.

    Conclusion

    TRY has anticancer effects on breast cancer cells by the induction of oxidative stress and apoptosis, as well as the regulation of genes involved in the process of mitochondrial apoptosis.

    Keywords: Breast cancer, Gene, miRNA, Doxorubicin
  • Amir Gholamzad, Melina Moulaeian, Mahsa Khatibi, Mohammadmatin Nourikhani, Mehrdad Gholamzad *, Yalda Goudarzi Page 5

    Cardiovascular disease (CVD) is a leading cause of death worldwide, and it has been found to have a strong genetic component. In recent years, there has been much interest in the role of microRNAs (miRNAs) in CVD. miRNAs are small non-coding RNAs that regulate gene expression post-transcriptionally by binding to the 3' untranslated region (UTR) of target mRNAs. Many studies have shown that miRNAs play a crucial role in various physiological processes, including the regulation of cellular functions involved in the development of CVD. Several miRNAs have been identified that are involved in the pathogenesis of CVD, and some of them are associated with specific cardiovascular risk factors, such as hypertension or diabetes. It has been suggested that targeting specific miRNAs or combinations of miRNAs could serve as a novel therapeutic approach for CVD. Moreover, studies have also shown that certain genes are involved in CVD risk and progression leading to different clinical manifestations like coronary artery disease, heart failure, and valvular disease. Some of these genes are involved in lipid metabolism, inflammation, and cell proliferation and differentiation, and their expression is regulated by miRNAs. In conclusion, a complex interaction between genes and miRNAs contributes to CVD pathogenesis, and further research is required to fully understand the mechanisms involved. Nevertheless, the identification of specific miRNAs that are involved in CVD provides potential targets for future therapeutics.

    Keywords: MicroRNA, CVD, Genes
  • Vahid Naseh *, Jalal Rezaeidian, Maliheh Entezari, Hakimeh Ziyadi, Mehrdad Hashemi Page 6
    Background

    Breast cancer (BC) is the second leading cause of death due to cancer among women worldwide. Therefore, the present study investigates the cytotoxic effects of piperine on the breast cancer cell line (MCF7) and the genes of the apoptotic pathway. 

    Objectives

    This research was performed to assess the effect of piperine on BC cells and the change in the expression level of bax gene through the induction of apoptosis.

    Methods

    MCF-7 cells were prepared by the Pasteur Institute, Iran. Cytotoxicity of piperine at concentrations of (5, 10, 15, 20, and 25 µM) during 24, 48, and 72 hours was evaluated by MTT assay. The cell apoptosis and bax gene expression were evaluated by Flow Cytometry and qReal-time PCR, respectively. Finally, the statistical analysis of MTT and RT- PCR data was done by SPSS software version 22. 

    Results

    The piperine showed concentration-dependent cytotoxic effects on the MCF-7 cell line in MTT assay. The bax gene expression level has a significant increase in piperine-treated cells compared to the untreated ones. The MCF-7 cell apoptosis at IC50 concentration of piperine was measured at 58.3% during 48-h treatment. 

    Conclusions

    In general, it can be concluded that piperine has cytotoxic effects against breast cancer by inducing apoptosis via overexpressing of bax.

    Keywords: Piperine, Breast Cancer, Apoptosis, MTT Assay, Flow Cytometry