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Showing 5 results for Fibroblast
Comparison of the expression levels of Fas and Apaf-1 genes in systemic sclerosis dermal fibroblasts
Majid Abed Khojasteh , Fereshteh Alsahebfosoul , Mahdi Mahmoudi , Mohammad Bagher Mahmoudi , Shayan Mostafaei , Mazdak Ganjalikhani-Hakemi , Farhad Gharibdoost ,
Volume 74, Issue 4 (7-2016)
Abstract

Background: Systemic sclerosis (SSc) is an autoimmune rheumatic connective tissue disease. In normal wound healing process, fibroblasts are activated, proliferated and involved in tissue repair, and then removed by apoptosis. In systemic sclerosis, patient’s fibrosis occurs when fibroblasts become resistant to apoptosis and secrete a large amount of collagen and other extracellular matrixes. As the primary causes the disease are very complex and often unknown, it is necessary to consider or target the secondary causes of disease, such as the unresponsiveness of activated fibroblasts to apoptosis as the major factor in the creation and deployment of illness. In this study, we examined the expression levels of two key pro-apoptotic genes, Fas and Apaf-1, which are respectively involved in external and internal pathway of apoptosis.

Methods: In a case-control study skin biopsy samples were obtained from 19 patients with diffuse SSc, and 16 healthy controls. Dermal fibroblasts were cultured and total RNA was isolated from cell populations using High Pure RNA Isolation Kit (Roche Applied Science, Mannheim, Germany), followed by cDNA synthesis using RevertAid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific Inc., Massachusetts, USA). Real-time PCR was performed using SYBRGreen gene expression master mix (Takara Shuzo, Co., Ltd, Shiga, Japan) and specific primers for Fas and Apaf-1. Real-time data were analyzed using the (2-ΔCT)×1000 method. Statistical analysis was accomplished by using the SPSS software, v22 (IBM, Armonk, NY, USA). The P value less than 0.05 were recognized as a significant threshold. All data are represented as the mean ± SEM.

Results: Our results showed no significant difference in Fas (P=0.8) and Apaf-1 (P=0.17) mRNA expression levels between skin fibroblasts of systemic sclerosis patients and healthy controls.

Conclusion: In this study we observed no significant change in Apaf-1 and Fas mRNA levels in systemic sclerosis fibroblasts compared to control group. Hence, Apaf-1 and Fas are not transcriptionally activated in SSc fibroblasts. Further studies need to take place on protein levels and function of these proteins to confirm the mRNA transcription results.


Estimation of isoform expression of PI3K and FGFR signaling pathway components by transcriptome analysis in bladder cancer
Zahra Ousati Ashtiani , Javad Tavakkoly-Bazzaz , Seyed Alireza Salami , Mohammad Reza Pourmand , Gholamreza Pourmand ,
Volume 75, Issue 6 (9-2017)
Abstract
Background: Aberrant pre-mRNA alternative splicing is a common event in cancer cells. Many abnormally spliced RNA variants have been observed in tumor cells and they can be used as biomarkers or therapeutic targets in new drug design. Increasing our knowledge in understanding the mechanisms of alternative pre-mRNA splicing for cancer-related genes and determination of cancer specific isoforms are important for the development of new strategies in cancer therapy. The aim of this study was isoforms identification and expression of PIK3CA, FGFR3 and FGFR1 genes in bladder cancer by RNA Sequencing and Real-Time PCR.
Methods: This cross-sectional study was conducted at Urology Research Center of Sina Hospital, Tehran University of Medical Sciences, Tehran, from September 2014 to October 2016. Paired tumor and adjacent normal tissues samples were obtained from 30 bladder cancer subjects. Total RNAs were extracted from bladder tumor and normal tissues. Quantitative and qualitative examinations have been done. After quality control, fragmentation of RNAs and cDNA library construction, next-generation RNA sequencing was performed. Resulting raw data were analyzed with different bioinformatics software. Differential expression was confirmed by Real-Time PCR.
Results: RNA sequencing results showed the number of PIK3CA (1 vs 3), FGFR3 (7 vs 6) and FGFR1 (9 vs 12) isoforms and their expression were different in bladder normal tissues in comparison to tumor tissues. Overexpression of PIK3CA gene have been observed in 42% of tumor samples but statistically was not significant. Increased FGFR3 gene (P=0.01) and decreased FGFR1 (P=0.01) expression were significant. There was an association with overexpression of FGFR3 and cigarette smoking ((P=0.037) and family history (P=0.004).
Conclusion: RNA sequencing make possible to do the accurate assessment of transcript abundance and identification of different isoforms resulted from aberrant pre-mRNA alternative splicing, which is a crucial process for the maturation of transcripts of multi-exon genes. Regarding the differences in isoforms expression in tumor and normal tissues of bladder cancer, they have potential to be used as biomarkers and sensitive targets for cancer therapy.

Anti-proliferative and pro-apoptotic effects of gallic acid on the breast adenocarcinoma cell lines SKBR3 versus normal fibroblast cells (HU-02)
Roghayeh Larki, Leila Rouhi , Seyed Hossein Hejazi ,
Volume 76, Issue 3 (6-2018)
Abstract
Background: Breast cancer is a malignant proliferation of epithelial cells that lining the ducts or lobules of the breast. Breast cancer is the second common cancer (after lung cancer) in women. Gallic acid, being a polyphenols, has been reported for its antiproliferative activity against many cancer cell lines. Objective of the present study is effect of gallic acid on proliferation and apoptosis of the human breast adenocarcinoma cell lines SKBR3 and normal fibroblasts cells.
Methods: This experimental study was performed in cellular and developmental biology of Shahrekord Islamic Azad University, Iran from April to August 2015. For anti-cancer activity, in this study SKBR3 cells and normal fibroblast cells (HU-02) were cultured in Dulbecco's modified eagle's medium, DMEM (Gibco, Life Technologies, Inc., New York, USA) medium with 10% fetal bovine serum, FBS (Gibco, Life Technologies, Inc., New York, USA). The SKBR3 and normal fibroblast cells were treated in the medium of DMEM medium and gallic acid (20, 40, 80, 100 and 200 µg/ml) for 24, 48 and 72 hours. Cells viability was assessed by MTS (Methyl- Thiazol-) assay. Cells were seeded at 5×103 cells/ml in 96 well plates and incubated for 24 hours. Then metabolites of bacteria were added, after indicated times MTS (20µl) was added and the absorbance was measured at 492 nm using ELISA plate reader. The percentage of apoptosis induction was determined by flow cytometry analysis using Annexin-V fluorescein isothiocyanate (FITC) kit (BioVision Products, CA, USA) in 20, 40, 80, 100 and 200 µg/ml concentration of gallic acid at 48 hours incubation.
Results: Gallic acid decreases significantly the viability of SKBR3 cell line in a time and dose dependent manner. So that the most effective concentration of this substance was 200 µg/ml and 72 hours after treatment (P< 0.05). According to the data of Annexin-PI, the highest apoptosis induction rate was seen in 200 µg/ml (P< 0.05). While gallic acid in various concentrations had no significant effect on normal fibroblast cells.
Conclusion: Objective of the present study is effect of gallic acid on proliferation and apoptosis of the human breast adenocarcinoma cell lines SKBR3 and normal fibroblasts cells.

Establishment and the importance of chicken pluripotent stem cells and their role in vaccine production: review article
Maryam Farzaneh, Mojgan Hosseini,
Volume 78, Issue 4 (7-2020)
Abstract
Chick embryos are a great historical research model in basic and applied sciences. Along with other animal models, avian and specifically chicken embryo has been attended, as well. Avian fertilized eggs as a natural bioreactor are an efficient tool for producing recombinant proteins and vaccines manufacturing. Due to the limitations of birds' eggs for viral replication, avian stem cells culture technologies access to safe methods as well as large-scale production of a variety of human and animal vaccines. Chicken pluripotent stem cells present the unique property of self-renewal and the ability to generate differentiated progeny in all embryonic lineages such as ectoderm, mesoderm, and endoderm in vitro. For the first time, chicken embryonic stem cells (cESCs) derived from the blastodermal cells of stage X embryos in vitro. Chicken ESC provides a great model of early embryo and they are useful for gene manipulation, virus proliferation, and the generation of transgenic birds. In addition to blastodermal cells, pluripotent cell lines can be produced by reprogramming of chicken fibroblasts into induced pluripotent stem cells (iPSCs) with transcription factors such as OCT4, NANOG, SOX2, KLF4, LIN28, and C-MYC that are well known to contribute to the reprogramming of somatic cells into an iPSCs. Similar to chicken ESCs, iPSCs have properties of unlimited self-renewal in vitro and the capacity for differentiation to all three embryonic germ layers. Chicken iPSCs have been a useful tool for the production of transgenic birds and viral vaccines. Despite the benefits and multiple applications of chicken pluripotent stem cells, the propagation of these cells is limited and some important challenges should be eliminated before their use in vaccine manufacturing. It is necessary to define the appropriate culture conditions for chicken pluripotent stem cells. For example, the presence of endogenous viruses in the avian species should be evaluated for human vaccine production. Currently, primary chicken fibroblast cells are still mainly used for vaccine production. This review covers the resources to achieve chicken derived cell lines for vaccine manufacturing.
 

Effect of resistance training modalities on angiogenic indices in sedentary male students
Mohammad Ali Gharaat , Yaghoob Mehri Alvar,
Volume 81, Issue 6 (9-2023)
Abstract
Background: Angiogenesis is a physiological process leading to capillary density enhancement and better blood distribution in skeletal muscles, which triggers in response to physical training. The present study aimed to investigate the changes in physiological factors involved in angiogenesis in response to circuit or traditional resistance training.
Methods: Thirty-six healthy sedentary students who were studying at Shahid Rajaee Teacher Training University of Tehran (age: 22.1±2.3 years; height: 172.7±5.1 cm) volunteered to participate in the study (from October 2021 to February 2022). Following a pre-test to evaluate one repetition maximum (1RM) of selected movements (Leg Press, Leg Curl, Leg Extension, Bench Pull, Seated Row, Biceps Curl), subjects randomly divided into Circuit Resistance training (CRT) (training protocol included 4 circles/3 times a week/8 weeks circuit performance/50-55% 1 Repetition Maximum (1RM), n=12), Traditional resistance training (RT) (training protocol included 8 repetition/3 set/3 time per week/8 weeks of same movements with 75% 1RM followed by 2 minutes break to rest between the sets; n=12) and the control group without any regular training (n=12). We assessed the level of Vascular Endothelial Growth Factor (VEGF), plasma level of growth hormone (GH), and Basic Fibroblast Growth Factor (BFGF) to the mentioned training methods. Data were evaluated by utilizing SPSS version 14.
Results: Present findings showed that CRT and RT protocols resulted in significant increases in post-test compared to pre-test in VEGF (P=0.00), GH (P=0.04), and BFGF (P=0.00). In addition, the magnitude of changes in VEGF and GH were significantly greater than the magnitude of changes in control group in post-test (P=0.03, and 0.001, respectively). Furthermore, there was a strong correlation between absolute values of GH and VEGF (r=0.74 and r=0.71) following CRT (P=0.01) and RT (P=0.02).
Conclusion: This study demonstrated that CRT and RT might enhance angiogenesis through an increase in VEGF, bFGF and GH, leading to better blood distribution in muscles.

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