فهرست مطالب

Reports of Biochemistry & Molecular Biology - Volume:8 Issue:3, 2020
  • Volume:8 Issue:3, 2020
  • تاریخ انتشار: 1398/10/19
  • تعداد عناوین: 11
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  • Zahra Taheri, Hamid Asadzadeh Aghdaei, Shiva Irani*, Mohammad Hossein Modarressi, Zahra Noormohammadi Pages 208-215
    Background

    Abnormal DNA methylation leading to altered transcription of certain genes occurs frequently in colorectal cancer (CRC). As with protein-coding genes, microRNAs (miRNAs) may be targeted for methylation in CRC; however, the methylation state of miRNA genes in CRC, especially in primary lesions, has not yet been completely elucidated. To understand the impact of DNA methylation on the miR-200c/141 cluster promoter, we investigated the methylation and expression of miR-141 in precancerous lesions and colorectal cancer.

    Methods

    In this cross-sectional study, 208 colorectal tissue samples, including 34 tumor tissue samples, 60 precancerous lesions with matched normal adjacent tissues, and 20 normal tissue samples, were collected. Promoter methylation of the miR-200c/141 cluster was studied using methylation-specific PCR. MiR-141 expression was examined using quantitative real-time PCR.

    Results

    Our findings showed that the miR-200c/141 cluster promoter region was most frequently hypermethylated in colorectal tumors and adenomatous polyps, but unmethylated in hyperplastic polyp tissues (P < 0.001). DNA methylation of the miR-200c/141 cluster and the tumor stage were significantly correlated (P = 0.002); however, miR-141 expression difference between the tumor and polyp samples was not significant (p = 0.6).

    Conclusions

    The DNA methylation status of the miR-200c/141 cluster could serve as a progression marker from benign polyps to colorectal cancer.

    Keywords: Colonic Polyps, Colorectal Cancer, DNA methylation, MiR-141
  • Mohammed Saeed Ali, Rasha Mohamed Hussein*, Mohamed Ahmed Kandeil Pages 216-226
    Background

    Selenium is a mineral that showed both pro- and anti-oxidant activities in various disease models. In this study, we evaluated the anti-tumor effect of selenium against 1,2-dimethylhydrazine (DMH)-induced colorectal cancer in BALB/C mice and its effect on apoptosis and angiogenesis.

    Methods

    Colorectal cancer was induced by subcutaneous injection of DMH (20 mg/kg body weight) in BALB/C mice once weekly for 20 weeks. Selenium (200 mg/L) was given to DMH plus selenium-treated group in the drinking water for the next 3 months.

    Results

    The DMH plus selenium-treated group exhibited significantly lower expression of cloned caudal-type homebox gene -2 (CDX-2) and vascular endothelial growth factor (VEGF) but higher caspase-3 expression level at p<0.001 compared to the DMH-treated group. Moreover, a decrease in the reduced glutathione content and glutathione peroxidase activity but an increase in the malondialdehyde content were observed at p<0.001. Both macroscopic and microscopic examination of the colorectal tissues confirmed the results.

    Conclusions

    The anti-tumor effect of selenium against an induced colorectal cancer in mice is attributed to its pro-oxidant, anti-angiogenic and apoptotic effects.

    Keywords: Angiogenesis, Apoptosis, Cancer, Mineral, Oxidative Stress
  • Bijan Soleymani, Ali Mostafaie* Pages 227-235
    Background

    Inclusion body formation in E. coli is a significant problem in recombinant protein production. The aim of this study was to improve the solubility of recombinant bovine sex determining region Y protein (SRY) in BL21 (DE3) E. coli cells.

    Methods

    In this research two recombinant bovine SRY (rbSRY) sequences were analyzed; these were wild-type SRY (wtbSRY) and codon-optimized SRY (cobSRY). Their expression in various culture conditions was examined; these differences included IPTG concentrations, temperatures, and media stabilizers.

    Results

    IPTG and temperature significantly affected rbSRY solubility (P < 0.001). The optimum IPTG concentration and temperatures for wtbSRY and cobSRY induction were 0.3 mM at 27 and 32 °C, respectively. In addition, arginine and sorbitol concentrations significantly affected rbSRY solubility (P < 0.01). Solubility of rbSRY protein was highest from the cobSRY construct in the presence 0.2 M arginine and 0.3 M sorbitol. The highest inclusion body production occurred with high glucose concentrations.

    Conclusions

    We found that modifications in temperature and IPTG and stabilizer concentrations affected rbSRY solubility.

    Keywords: Cobsry, Inclusion Bodies, Recombinant Bovine SRY Protein, Solubility, Wtbsry
  • Hoda Nadimi, Abolghassem Djazayery, Mohammad Hassan Javanbakht, Ahmadreza Dehpour, Ehsan Ghaedi, Hoda Derakhshanian, Hamed Mohammadi, Mahnaz Zarei, Mahmoud Djalali* Pages 236-243
    Background

    Diabetes mellitus and metabolic disorders are a major burden on the healthcare system. Irisin is a novel myokine reported to have beneficial effects on glucose and lipid metabolism. Vitamin D deficiency has been implicated in the development of diabetes and hold a critical role in diabetes-related complications. In the present study, we examined the efficacy of vitamin D supplementation on serum irisin levels, skeletal muscle irisin levels, and the expression of the irisin precursor, FNDC5 (fibronectin-type III domain-containing 5) in type I diabetes mellitus rats.

    Methods

    Thirty-six adult male Sprague-Dawley rats (150 – 250 g) were randomly divided into four groups: group I: healthy control rats with no treatment (n=8), group II: healthy control rats receiving sesame oil as a placebo (n=8), group III: diabetic rats receiving sesame oil as placebo (n=10), group IV: diabetic rats treated with 4300 IU/kg/week vitamin D (n=10). Diabetes was induced by intraperitoneal (IP) injection of streptozotocin. At the end of the vitamin D intervention blood and triceps muscle samples were collected. RNA was extracted from muscle and real-time PCR was performed to examine FNDC5 gene expression.

    Results

    Our study showed that the administration of vitamin D (4300 IU/kg/week) in a streptozotocin-diabetic rat model resulted in increased serum vitamin D levels, FNDC5 gene expression and muscle irisin levels. However, the levels of serum irisin were not significantly changed by the administration of vitamin D.

    Conclusions

    In conclusion, we show that vitamin D supplementation enhances serum vitamin D levels, FDNC5 gene expression and muscle irisin levels in the streptozotocin-diabetic rat model. Our study highlights the potential therapeutic effect of vitamin D supplementation for diabetes mellitus.

    Keywords: Diabetes, FNDC5, Irisin, Vitamin D
  • Fatemeh Mohammad Rezaei, Shahryar Hashemzadeh, Reyhaneh Ravanbakhsh Gavgani, Mohammadali Hosseinpour Feizi, Nasser Pouladi, Hossein Samadi Kafil, Leila Rostamizadeh, Vahid Kholghi Oskooei, Mohammad Taheri, Ebrahim Sakhinia* Pages 251-259
    Background

    Colorectal cancer (CRC) is one of the most commonly-diagnosed malignancies throughout the world and the fourth-leading cause of cancer deaths globally. Angiogenesis and the resultant tumor neovascularization is a well-known cancer hallmark. Here we investigated the expression of FLT1 and KDR, the influential genes in angiogenesis regulation, in CRC patients.

    Methods

    We assessed FLT1 and KDR mRNA expression in 47 CRC samples and matched adjacent non-cancerous tissues (ANCT) by quantitative real-time PCR. The Spearmen correlation coefficient and receiver operating characteristic (ROC) curves were also examined.

    Results

    Both genes were expressed at significantly greater levels in CRC tissues than in ANCT (p < 0.05). A significant association was found between KDR expression and disease stage and lymph status in CRC patients. Furthermore, the Spearman correlation demonstrated a moderate correlation between FLT1 and KDR expression in CRC samples. Finally, ROC curve analysis demonstrated that FLT1 had the greatest sensitivity (85.1%), while the greatest specificity was achieved by a combination of the two genes.

    Conclusions

    The dysregulated FLT1 and KDR expression, in addition to the observed correlation and ROC curve results, indicate the critical importance of angiogenesis among the cancer pathways in CRC. These data can broaden our current knowledge of angiogenesis in CRC to improve disease diagnosis and patient treatment.

    Keywords: Colorectal cancer, FLT1, KDR, Gene expression
  • Revathi Panduranga Shenoy, Gagana Hanumaiah, Varashree Bolar Suryakanth*, Priyanka Shridharan Pages 260-266
    Background

    The study aims at the comparison and correlation of serum levels of fructosamine and erythrocyte Na+/ K+ ATPase in Gestational diabetes mellitus (GDM) and Non Gestational Diabetes Mellitus (Non GDM).

    Methods

    A total of 326 samples were divided into 4 groups. Pregnant women between the age group of 20-40 years who gave samples for Oral Glucose Tolerance Test (OGTT) were included as the subjects. Anonymized and left over fasting and 2 hours’ samples were collected from biochemistry laboratory, Kasturba Hospital, Manipal.

    Results

    In the comparison of fructosamine levels in GDM and Non GDM, fructosamine was found to be significant (p value<0.001) in both fasting and 2 hours (G2) blood glucose condition. Na+/ K+ ATPase did not show any significant variation between the groups. Correlation was not significant between the parameters.

    Conclusions

    Fructosamine showed significant increase when compared between the groups, whereas significant correlation is not obtained between the parameters. Thus, the use fructosamine as a diagnosis tool becomes inconclusive. Further studies must be carried out to identify a marker which reduces the interferences observed in fructosamine and to find out the exact relationship between hyperglycaemia and Na+/ K+ ATPase activity.

    Keywords: Fructosamine, Gestational Diabetes Mellitus, Na+, K+ Atpase Enzyme Activity, Screening Test
  • Motahare Sadat Hosseini, Fatemehsadat Hosseini, Abdolreza Ahmadi, Masoud Mozafari, Issa Amjadi* Pages 260-268
    Background

    In recent years, prostate cancer prevails as one of the lead cancers affecting men. Currently, prostate cancer research involves the phytochemical study of plants with anti-tumour effects. This study compares the anti-tumour effects of three plant species indigenous to Iran and their interaction with cluster of differentiation (CD)-82 protein, a therapeutic target found in prostate cancer cells.

    Methods

    The extracts of Hypericum perforatum, Achillea millefolium, and Aloe vera were prepared and their toxicological, cellular and gene expression responses were evaluated in PC-3 human prostate cancer cells and normal human chondrocyte cell line C28/I2. They were exposed to different concentrations of the plants (10 mg/mL, 5 mg/mL, 1 mg/mL, 100 µg/mL, 10 µg/mL, and 1 µg/mL) at three exposure time points (24, 48, 72 hours) to determine cancer cell cytotoxicity and gene expression profiles.

    Results

    Half-maximal inhibitory concentration (IC50) in PC-3 cells ranged from 0.6 to 8.5 mg/mL for H. perforatum extract, from 0.4 to 7.5 mg/mL for A. Millefolium extract, and from 0.2 to 8.0 mg/mL for A. vera extract in a time-dependent manner. A. vera extract caused the highest cell death levels in PC-3 cells (94%) and C28/I2 cells (57%) after 48 hours. A 1.97-, 3.00-, and 3.48-fold increase in relative gene expression of CD82 was observed for H. perforatum, A. millefolium, and A. vera extracts, respectively

    Conclusions

    A. vera and A. millefolium extracts are a selective inhibitor of prostate cancer cells and a potent activator of CD82 expression.

    Keywords: CD82, Gene expression, Herbal medicine, Prostate cancer
  • Mustafa Kahtan Al Bayaty*, Salma Abdul, Rudha Abass, Mohammed Faraj Al Marjani Pages 269-277
    Background

    Gastric cancer is still the main health threat being the third leading cause of deaths from cancers in the world. The major risk behind the gastric cancer is that it remains asymptomatic in the early stages and in (97%) cases it metastasizes to other organs. Gastric cancer is a multifactorial disease in which Helicobacter pylori (H. pylori) has been known as a risk factor. However, patients with gastritis, especially atrophic gastritis and gastric ulcer have been shown to be at an increased risk for developing gastric cancer.

    Methods

    This study included measuring the serum levels of E-Cadherin protein, carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) in 30 patients diagnosed with gastritis, 20 gastric ulcer patients, 20 gastric cancer patients and in 20 healthy volunteers serving as the control group.

    Results

    Infection with H. pylori was diagnosed by serology (IgA and IgG antibodies) as well as by rapid urease test (RUT) and histology. The results showed that 50 (71.4%) of the patients were positive for H. pylori. Levels of E-Cadherin were increased significantly in all patients in comparison to the control group with a large significant increase in the gastric cancer group. The levels of E-Cadherin were also significantly increased in H. pylori infected patients compared to H. pylori negative patients. A non-significant difference in the levels of CA19-9 and CEA was observed in all patients in comparison to healthy controls.

    Conclusions

    This study concluded that serum E-Cadherin could be considered as a potential marker in diagnosis of gastric cancer.

    Keywords: E-Cadherin, Gastric Cancer, Gastric Ulcer, Gastritis, Helicobacter Pylori
  • Ali Jumma Kareem, Jehan Abdul Sattar Salman* Pages 278-286
    Background

    Dextran is a commercially available bacterial exopolysaccharide (EPS) with several industrial applications in the food industry and in the biomedical industry as an adjuvant, emulsifier, carrier, and stabilizer. The production of dextran at the industrial level occurs through the fermentation of a sucrose-rich medium. Research to optimize dextran production has found that the yield of dextran varies depending the specific conditions for production. The aim of this study was to produce dextran and establish the optimal conditions for dextran biosynthesis from different Lactobacillus species isolated from healthy vaginal and infant stool samples.

    Methods

    Lactobacillus spp. were isolated and identified from vaginal and infant stool samples via the VITEK 2 system. The presence of dextran biosynthesis from the different Lactobacillus spp. isolates was determined by a screening test for mucoid colonies and confirmed via theethanol precipitation method. To optimize for the maximum yield of dextran, the effects of various parameters such as temperature, incubation time, pH, inoculum size, aeration, and sucrose concentration were examined.

    Results

    All Lactobacillus spp. isolates were able to produce dextran. The optimal conditions for dextran production was at 24 hours of incubation at 30 °Cwith 15% sucrose, 4% inoculation size at pH 7.0 in aerobic conditions. This yielded a dextran dry weight of 580 mg/100 mL.

    Conclusions

    Dextran production from Lactobacillus species isolates vagina and infant stool had the ability to produce dextran.

    Conclusions

    The DNA methylation status of the miR-200c/141 cluster could serve as a progressionmarker frombenignpolypstocolorectalcancer.

    Keywords: Dextran, Lactobacillusspp, Optimum Conditions, Precipitation
  • Ola Mostafa Tork, Laila Ahmed Rashed, Nermeen Bakr Sadek, Marwa Sayed Abdel Tawab* Pages 287-300
    Background

    Neuroprotective mechanisms triggered by peroxisome proliferator-activated receptor-gamma agonist: pioglitazone (PIO) and glucagon-like peptide 1 analog: exendin-4 (Ex-4) in neurological diseases were reported, but whether mitochondrial biogenesis is involved or not in their neuro-protective mechanisms in type 1 Diabetes Mellitus (T1DM); has not been studied before. To bridge this gap, we investigated the effect of PIO and Ex-4 on brain mitochondrial biogenesis in streptozotocin-induced diabetes in rats.

    Methods

    Seven weeks after induction of diabetes in rats, serum fasting glucose and insulin were measured in studied groups. The brain was removed for histological analysis and assessment of: mitochondrial complexes I and II, ATP, H2O2, brain derived neurotrophic factor (BDNF), cytochrome c and hemeoxygenase (HO)-1 activity, and relative gene expression of the nuclear factor; Nrf2and the apoptotic markers: bax& bcl2and mitochondrial biogenesis markers; peroxisome proliferator–activated receptor γ coactivator (PGC) 1-α and sirtuin 1 (SIRT-1) and AMP-activated protein kinase (AMPK) and c-Jun-N-terminal kinase (JNK) proteins.

    Results

    Brain in untreated rats showed neurodegeneration area and significantly rising H2O2and JNK, up-regulation of bax, down-regulation of bcl2. These changes were paralleled with significant reduction in Nrf2, HO-1, BDNF, complex I, II and ATP and SIRT-1/ PGC1-αexpression. PIO and Ex-4 significantly improved the reported changes. Combined modality showed better improvement relative to each drug alone.

    Conclusions

    PIO and Ex-4 may have neuroprotective effects in T1DM, via targeting altered mitochondrial biogenesis probably due to modulation of brain SIRT-1signaling, improvement of oxidative stress and equilibrating the balance between pro-apoptotic and anti-apoptotic mediators.

    Keywords: Brain derived neurotrophic factor, Diabetic neurodegeneration, Exendin-4, oxidative stress, PGC1-α
  • Mahboubeh Poursiyami, Foroogh Nejatollahi*, Afagh Moatari Pages 301-309
    Background

    Immunotherapies using monoclonal antibodies against influenza A hemagglutinin (HA) has been an effective means for controlling Influenza spread. An alternative method for viral prophylaxis and treatment is the development of human single-chain variable fragment (scFv) antibodies with no human anti-mouse antibody (HAMA) response and high specificity. In the present study, two highly conserved sequences of HA were used to select specific neutralizing scFvs againstH3N2 strain of influenza A virus.

    Methods

    Biopanning process was performed to isolate specific scFv antibodies against highly conserved HA sequences, aa173-181 and 227-239, of the influenza A H3N2 strain from a scFvlibrary. The peptide-binding specificity of the selected clones was examined via phage ELISA. The soluble forms of the clones were prepared and assessed using western blot analysis and neutralization efficiency of the selected clones were examined by TCID50 neutralizing assay and real-time PCR.

    Results

    scFv 1 and scFv 2 were selected against HA of H3N2 influenza A virus with frequencies of 95% and 30% in the panning process, respectively. Western blot analysis confirmed the scFv band size. Significant neutralization in the presence of scFv 1 and scFv 2 were obtained. Real time PCR revealed significant decrease in viral copy number.

    Conclusions

    Two specific neutralizing scFvsagainst two highly conserved neutralizing epitopes of the influenza A virus HA glycoprotein were selected. A strong neutralization effect of scFv1, showed the potential of this antibody for H3N2 influenza A controlling in the viral spread

    Keywords: Hemagglutinin, H3N2 Influenza A, Neutralization, scFv