Jentashapir Journal of Cellular and Molecular Biology
Year:2021 | Volume:12 | Issue:3|Papers Count:8

Background: Gastric cancer is the second reason for cancer mortality worldwide, with a high capacity for metastasis. Long noncoding RNAs (lncRNAs) are recently described as lengthy transcripts with no open reading frame. The lncRNAs play an important role in critical cellular and molecular pathways, including cell cycle, growth, differentiation, and apoptosis. Therefore, it is not surprising that abnormal expression of lncRNAs may be involved in human cancers. The HOX antisense intergenic RNA (HOTAIR) is a highly cited lncRNAs whose altered expression has been reported in a variety of humancancers such as gastric cancer. Epithelial to mesenchymal transition (EMT) is a cellular route in which an epithelial phenotype of the cells can be changed into the mesenchymal state. The signaling pathways involved in EMT are related to cancer metastasis and recurrence of gastric cancer. Methods: The present study was aimed to investigate the effect of HOTAIR gene silencing on expression levels of fibronectin 1 (FN1) and claudin-4 (CLDN4) genes, two important markers of EMT, in AGS cellular model of gastric cancer. The AGS cells were exposed to the HOTAIR-specific siRNA for 48 hours. The extracted RNAs were subjected to complementary DNA synthesis and real-time PCR. Data were analyzed using 2-

Ct method. Cells with no siRNA treatment were considered the control set. The P-value < 0. 05 was considered statistically significant. Results: The observed data showed that the expression levels of two EMT markers FN1 and CLDN4, were significantly decreased after HOTAIR silencing. Conclusions: This study demonstrates that HOTAIR can regulate the EMT signaling pathway through critical EMT factors like FN1 and CLDN4 transcripts. However, a long way remains to apply this finding in therapeutic approach, and further experiments are needed.

Background: In recent years, the relationship between cancer cells and electromagnetic radiation has received much attention. Objectives: The present study aimed to evaluate the effects of different intensities of electromagnetic fields on gastric cancer cell lines (AGS). Methods: After preparing AGS and Hu02 (normal) cell lines, they were exposed to magnetic flux densities of 0. 25, 0. 5, 1, and 2 millitesla (mT) for 18 h. The cell viability was studied by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The expression levels of hes1 and hsa-circ-0068530 RNAs were studied by the quantitative Real-time-PCR technique. Results: The inhibition of gastric cancer cell line growth was observed under the influence of electromagnetic fields at different intensities. However, they did not affect the viability of normal cells. A sharp increase in the expression of hes1 and hsa-circ-0068530 genes was observed in normal cells exposed to 2 mT electromagnetic fields. Conclusions: In general, it can be concluded that the effect of electromagnetic fields on gastric cancer cells depends on their intensity. Magnetic flux densities of 0. 25 and 0. 5 mT had anti-cancer effects and magnetic flux density of 2 mT showed carcinogenic effects.

Background: Ovarian cancer is the deadliest gynecologic cancer. Studiesonthe therapeutic properties of Ginkgo bilobaand flunixin showed that these drugs, singly or in combination with other drugs, have anti-cancer activities. Different genes are involved in apoptosis regulation. The BIM gene is one of the most important regulators of this process. BIM has different roles, including cell cycle regulation, apoptosis induction, deoxyribonucleic acid recombination, chromosomal segregation, and cell aging. Methods: This study evaluated the viability percentage of the A2780s cell line with Ginkgo biloba and flunixin at different concentrations, compared to that of the control group. Then, the half-maximal inhibitory concentration (IC50) values of Ginkgo biloba and flunixin were determined within 24 h. Then, the expression of the BIM gene was evaluated using a real-time polymerase chain reaction (PCR). Results: The IC50 results showed that Ginkgo biloba and flunixin significantly reduced cell life (P < 0. 01) depending on time and concentration. The results of real-time PCR showed that cell treatment with Ginkgo biloba and flunixin significantly increased BIM expression. Conclusions: The results of this experiment indicated that BIM gene expression was increased in cancer cells treated with Ginkgo biloba and flunixin, compared to that reported for control cells. Therefore, with further research in the future, these compounds can be used for the development of ovarian anti-cancer drugs.

Objectives: Identifying the effective exercise protocol that attenuates the functional and molecular disturbances in different regions of the brain, in particular the cerebellum, can help the proper management of neuropathies in diabetic patients. Methods: Twenty rats were randomly divided into four groups: (1) Normal control group (CON), (2) normal exercise group (TH), (3) diabetes control group (DC), and (4) diabetes exercise group (TD). Diabetes was induced by i. p injection of a single dose of streptozotocin (50 mg/kg). The endurance training protocol was performed on a treadmill for five days a week for six weeks with moderate intensity. The activities of antioxidant enzymes and the expression or release of apoptotic factors were analyzed based on data from rat cerebellum tissue at the end of the experiments. Results: Six weeks of endurance training improved the oxidative defense system by increasing the activities of SOD (from 3. 70  0. 64 to 6. 55 0. 56), GPx (from 3. 42 0. 73 to 4. 84 0. 62), and catalase (from 1. 36 0. 23 to 3. 59 0. 37) and reducing the MDA concentration (from 6. 81 1. 34 to 4. 33 1. 03) in the cerebellum of diabetic rats. Increased expression or cytosolic release of apoptotic effectors such as bax, caspase 3, and cytochrome c in the cerebellum of diabetic rats were attenuated following exercise training. Conclusions: Our research results showed that six weeks of endurance training may be helpful for the attenuation of neuropathies in diabetic patients by the attenuation of apoptosis and oxidative stress in the cerebellum.

Background: In vitro biofilm formation of H. pylori is demonstrated; however, its potential role in the persistent infection of the human stomach has not yet been addressed. Objectives: The aim of this study was to assess the biofilm formation of clinical H. pylori isolates on an epithelial cell line, a line that produces mucin. Methods: H. pylori isolates consisting of an efficient (19B) and a weak (4B) biofilm formation ability, were selected from screening of the clinical isolates. Their adhesion index was determined after 2h incubation with the semi-confluent monolayers of MKN-45 cells. Their biofilm formation was evaluated after 24 and 72hincubation with MKN-45 cells using amodified adherence assay developed in this work. Production of biofilmwasquantitatively assessed byCFUenumerationandqualitatively by the immunofluorescence, and scanning-electron-microscopic (SEM) methods. Due to the importance of mucin in the binding of H. pylori and biofilm formation, the binding strength of the mucin binding protein, MUC5AC, and MUC1 with docking was investigated using cluspro webserver. Results: Using MKN-45 epithelial cell line as a model, significant differences were observed between the adhesion index of 19B and 4B isolates. After 24h, both isolates were able to form biofilms with significantly higher numbers of CFU for the 19B isolate. These results were confirmed by immunofluorescence and SEM such that after 24h, a cluster of coccoid bacteria on the MKN-45 cells in the form of microcolonies was observed. The docking results showed that MUC5AC demonstrated the most favorable interaction with H. pylori urease and BabA with docking energy scores of-931. 1 and-906. 3 kcal. mol-1, respectively. Conclusions: By developing an appropriate in situ biofilm assay, we investigated biofilm formation by clinical H. pylori isolates on the MKN-45 epithelial cell line. The establishment of such an in-situ model for studying the biofilm formation ability of clinical isolates can also be used to study cell-bacteria interactions in the context of a complex biofilm and also as a model for drug screening applications.

Background: Among the known ABL mutations in chronic myeloid leukemia (CML), T315I is of particular importance. The T315I mutation may develop resistant cells that increase disease progression. TWIST-1 expression is impaired in patients with increased drug resistance. Objectives: The current study aimed to measure the expression of TWIST-1 gene in CML patients to investigate its association with T315I mutation. Methods: Peripheral blood samples were taken from 40 CML patients. The expression of TWIST-1 and BCR-ABL1 genes was quantified by real-time polymerase chain reaction (PCR). The gene expression was evaluated by REST software. cDNA was used for amplification refractory mutation system (ARMS)-PCR reaction. Results: Of the 40 patients (age range: 19-72 years) participating in the study, 23 (57. 7%) were female, and 17 (42. 5%) were male. The expression of TWIST-1 gene was 43 184. 09-fold. The T315I mutation was detected in 3 (7. 5%) patients. Conclusions: According to our results, the TWIST-1 gene expression in patients with T315I mutation was significantly higher than patients without that mutation.

Background: Activated hepatic stellate cells (HSCs) are the primary mediators in the progression of hepatic fibrosis. The activation of toll-like receptor 4 (TLR4) signaling leads to the downregulation of the transmembrane inhibitory transforming growth factorbeta (TGF-) pseudoreceptor BMP and activin membrane-bound inhibitor (BAMBI) on HSCs. Fibroblast growth factor 21 (FGF21) is a natural secretory protein in the body with effects, such as the reduction of fat accumulation and oxidation of lipids; however; no study has investigated FGF21 ability to prevent the progression of liver fibrosis. Objectives: This study aimed to examine the beneficial effects of FGF21 to reduce cholesterol-activated human HSCs. Methods: The human HSCs were incubated in media containing different concentrations of cholesterol, including 25, 50, 75, 100, 125, and 150 M, for 24 h and then incubated with FGF21 for 24 h. Total ribonucleic acids were extracted and reversely transcribed into complementary deoxyribonucleic acid. A quantitative real-time polymerase chain reaction was performed in this study. Results: The results showed that the messenger ribonucleic acid (mRNA) expression of TGF-, collagen, type I, alpha 1 (collagen1), and TLR4 genes increased significantly in the presence of cholesterol (75 and 100 M), compared to that of the control group (* P < 0.05, ** P < 0.01, and *** P < 0.001); nevertheless, the mRNA expression of the BAMBI gene significantly reduced, compared to that of the control group (* P < 0.05). The FGF21 significantly reduced the mRNA expression of TGF-, collagen1, and TLR4 genes (# P < 0.05). The mRNA expression of the BAMBI gene significantly increased with FGF21 (# P < 0.05). Conclusions: It was concluded that the treatment with FGF21 reduces the cholesterol-activated HSCs by decreasing the mRNA expression of the TLR4, TGF-, and collagen1 genes and increasing the mRNA expression of the BAMBI gene.

Objectives: This study aimed to evaluate the impact of resistance exercise and donepezil on some neurotrophins gene expression and Trk receptors in the hippocampus of rats with Alzheimer’ s disease (AD). Methods: In this study, 32 male adult Wistar rats (mean weight: 230-280 g) were assigned into two groups of AD and control. The control and AD groups received normal saline and streptozotocin (STZ) through intraventricular injection, respectively. Then, six subgroups were considered: (1) control rest (Con); (2) control exercise (Con-Exe); (3) Alzheimer’ s rest (Alz); (4) Alzheimer’ s exercise (Alz-Exe); (5) Alzheimer’ s donepezil (Alz-Don); and (6) Alzheimer’ s donepezil-exercise (Alz-Don-Exe). Donepezil was fed daily at a dose of 1. 5 mg/kg to the treated groups. The three subgroups of exercising rats received exercises for eight weeks (three times a week). Each day, the resting groups were managed to decrease stress impacts. Twenty-four hours after the last session of exercise by the eighth week, deep anesthesia was applied, and the rats’ heads were severed. Results: Considering an error rate below 5% (P < 0. 05) and a confidence of more than 95%, a significant difference was observed in BDNF, NT3, NGF, TrkA, and TrkB values between exercising and donepezil-exercise rats compared to AD group. These values were considerably greater for donepezil-exercising Alzheimer’ s group. Besides, the donepezil group was not significantly different from the Alzheimer’ s group. Conclusions: Although the use of donepezil alone did not significantly increase the expression of the studied genes, the concomitant use of the drug and resistance training significantly increased the expression levels.