Cell Journal (Yakhteh)
Volume:24 Issue: 6, Jun 2022

Long non-coding RNAs (lncRNAs) feature prominently in tumors. Reportedly, lncRNA zinc finger E-box-binding homeobox 2 antisense RNA 1 (ZEB2-AS1) is aberrantly expressed in a variety of tumors. The present study was aimed to explore ZEB2-AS1 functions and determine mechanism in hepatocellular carcinoma(HCC) progression. In this experimental study, expressions of ZEB2-AS1, microRNA (miR)-582-5p and forkhead box C1 (FOXC1) mRNA in HCC tissues and cell lines were detected via quantitative reveres transcription polymerase chain reaction (qRT-PCR). After establishing gain- and loss-of-functions models, cell counting kit-8, 5-bromo-2’-deoxyuridine (BrdU), Transwell assays and flow cytometry analysis were conducted to examine HCC cell multiplication, migration, invasion and apoptosis, respectively. The targeted relationship between miR-582-5p and ZEB2-AS1 was verified via dual-luciferase reporter gene assay. Western blot was utilized for detecting FOXC1 expression in HCC cells after selectively regulating ZEB2-AS1 and miR-582-5p. In HCC tissues and cells, ZEB2-AS1 expression was increased. High ZEB2-AS1 expression was related to relatively large tumor volume, increased tumor-node-metastasis (TNM) stage and positive lymph node metastasis of the patients. ZEB2-AS1 overexpression facilitated HCC cell multiplication, migration, invasion and suppressed apoptosis, while ZEB2-AS1 knock-down caused the opposite effects. It was also confirmed that ZEB2-AS1 could competitively bind with miR-582-5p to repress its expression, and indirectly up-regulate FOXC1 expression level in HCC cells. The current study revealed that ZEB2-AS1 was over-expressed in HCC tissues and cells. It also upregulated FOXC1, through sponging miR-582-5p, to promote HCC progression. This provides new perspectives for elucidating the pathogenesis of HCC.

This study is aimed to explore the biological function of long intergenic non-protein coding RNA 265 (LINC00265) in hepatocellular carcinoma (HCC) cells, and evaluate its potential as a biomarker. GEPIA database and Kaplan-Meier Plotter database were employed to analyze LINC00265 expression in HCC tissue samples and its predicting value for prognosis. LINC00265 expression in HCC tissues and cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). After LINC00265 was overexpressed and knocked down in HCC cells, cell counting kit-8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (EdU) assays were adopted to detect the proliferation of HCC cells. Transwell assay was used to detect the migration and invasion of HCC cells. The interaction between LINC00265 and E2F transcription factor 1 (E2F1) was verified by the catRAPID online analysis tool, RNA pull-down experiment, and RNA binding protein immunoprecipitation (RIP) assay. The binding of E2F1 to the promoter region of cyclin-dependent kinases 2 (CDK2) was detected by dual-luciferase reporter assay and chromatin immunoprecipitation. The regulatory effects of LINC00265 and E2F1 on CDK2 expression were probed by Western blot. LINC00265 expression was increased in HCC tissues and cells. LINC00265 overexpression promoted the proliferation, migration, and invasion of HCC cells, and knocking down LINC00265 worked oppositely. LINC00265 could bind to E2F1, and LINC00265 could enhance the combination of E2F1 and CDK2 promoter regions, thus promoting CDK2 transcription. LINC00265 overexpression promoted the expression of CDK2 in HCC cells. Our data suggest that LINC00265 can promote the malignant behaviors of HCC cells by recruiting E2F1 to the promoter region of CDK2.

Non-small cell lung adenocarcinoma (NSCLC) is the most common type of lung cancer, which is considered as the most lethal and prevalent cancer worldwide. Recently, molecular changes have been implicated to play a significant role in the cancer progression. Despite of numerous studies, the molecular mechanism of NSCLC pathogenesis in each sub-stage remains unclear. Studying these molecular alterations gives us a chance to design successful therapeutic plans which is aimed in this research.

In this bioinformatics study, we compared the expression profile of 7 minor stages of NSCLCadenocarcinoma, including GSE41271, GSE42127, and GSE75037, to clarify the relation of molecular alterations and tumorigenesis. At first, 99 common differentially expressed genes (DEG) were obtained. Then, functional enrichment analysis and protein-protein interaction (PPI) network construction were performed to uncover the association of significant cellular and molecular changes. Finally, gene expression profile interactive analysis (GEPIA) was employed to validate the results by RNA-seq expression data.

Primary analysis showed that BMP4 was downregulated through the tumor progression to the stage IB andGPX2 was upregulated in the course of final tumor development to the stage IV and distant metastasis. Functional enrichment analysis indicated that BMP4 in the TGF-β signaling pathway and GPX2 in the glutathione metabolism pathway may be the key genes for NSCLC adenocarcinoma progression. GEPIA analysis revealed a correlation between BMP4 downregulation and GPX2 upregulation and lung adenocarcinoma (LUAD) progression and lower survival chances in LUAD patients which confirm microarray data.

Taken together, we suggested GPX2 as an oncogene by inhibiting apoptosis, promoting EMT and increasing glucose uptake in the final stages and BMP4 as a tumor suppressor via inducing apoptosis and arresting cell cycle in the early stages through lung adenocarcinoma (ADC) development to make them candidate genes to further cancer therapy investigations.

Osteoporosis is regarded as a silent disorder affecting bone slowly, leading to an increased risk of fractures. Lately, selenium has been found to be associated with the acquisition and maintenance of bone health by affecting the bone remodeling process. However, the mechanism of action of selenium on bone is poorly understood. Here, the protective effects of sodium selenite on the differentiation process of osteoblasts as well as under oxidative stress-induced conditions were evaluated. MC3T3-E1 cells, were treated with a various doses (0, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2, 6.4 ug/ml) of sodium selenite. Cell viability and cytotoxicity were observed by MTT and LDH assay. The osteogenic activity and expression level of osteogenic markers were confirmed through ALP activity, real-time RT-PCR and sirius red staining. Role of sodium selenite and involvement of WNT signaling was assessed by Axin-2 reporter assay and western blotting. It was observed that the sodium selenite could promote the ALP activity and collagen synthesis in pre-osteoblasts. Moreover, sodium selenite increased the mRNA expression levels of osteogenic transcriptional factors, such as runt-related transcription factor 2 (Runx2) and osterix (OSX). In addition, the terminal differentiation markers, such as osteocalcin (OCN), and collagen 1α (Col1α), were also increased. Treatment of sodium selenite recused the H2O2-induced inhibition of osteoblastic differentiation of pre-osteoblasts cells. Furthermore, sodium selenite restored the H2O2 repressed β-catenin stability and axin-2 reporter activity in MC3T3-E1 cells implicating involvement of WNT signaling pathway. It may be concluded that selenite can stimulate bone formation and rescue the oxidative repression of osteogenesis by activating WNT signaling pathway and may act as a potential therapeutic intervention for osteoporosis.

Autologous transplantation of epidermal cells has been used increasingly to treat vitiligo patients and is a simple, safe, and relatively efficient method. However, the outcome is not always satisfactory, and some patients show less or no response to this treatment. This study was evaluated to identify genes expressed differently among responders and non-responders to cell transplantation to find potential markers that could predict 'patients' responses to this type of cell therapy. Eleven stable vitiligo patients who received autologous epidermal cell transplantation were included in this clinical trial study. Before cell transplantation, skin samples were obtained from the recipient’s vitiligo lesions. After epidermal cell transplantation, patients were followed for at least six months to assess the response to epidermal cell injection. RNA sequencing was used to determine potential gene expression profile differences between responder and non-responder vitiligo patients. The RNA sequencing results showed differences in expression levels of 470 genes between the skin specimens of responder versus non-responder patients. There were 269 up-regulated genes and 201 down-regulatedgenes. Upregulated genes were involved in processes, such as Fatty Acid Omega Oxidation. Down-regulated geneswere related to PPAR signaling pathway, and estrogen signaling pathway. Among the most differentially expressed genes (DEGs) with the most altered RNA expression levels in responders versus non-responder patients, we selected three genes (up-regulated genes KRTAP10-11 and down-regulated genes IP6K2 and C9) as potential biomarkers, which are involved in associated pathways. Based on our findings, it is estimated that proposed genes might predict the response of vitiligo patients to cell therapy. However, further studies are required to clarify the role of these genes in pathogenesis and to characterize gene expression in a larger number of vitiligo patients in the context of epidermal cell transplantation therapy (registration number: IRCT201508201031N16).

Transient receptor potential vanilloid 1 (TRPV1) is a heat-activated nonselective cation channel that playsimportant role in the spermatogenesis, capacitation, acrosome reaction and sperm/oocyte fusion. Considering the hightesticular temperature and oxidative stress in varicocele condition, we aimed to assess expression of TRPV1 in sperm of infertile men.

In this case-control study, twenty-five men with varicocele (grade II and III) as well as twentyfivefertile were recruited. Sperm parameters, protamine deficiency (Chromomycin A3), DNA damage (TUNEL), lipid peroxidation (BODIPY), TRPV1 gene expression (real time polymerase chain reaction), TRPV1 protein (flowcytometry and immunocytochemical techniques), and acrosome reaction were assessed between fertile and varicocele groups.

We observed a significant decrease in the sperm parameters, and also, an increased DNA damage, lipid peroxidation, and protamine deficiency in varicocele group. Although, the mRNA expression of TRPV1 was similar between varicocele and fertile groups, its expression at the protein level was significantly decreased in the varicocele group in comparison with fertile group. Additionally, the TRPV1 localization was changed from the equatorial region to the acrosomal region of the head, especially in the acrosomal region, which was more significant in the fertile group than the varicocele group after inducing acrosome reaction.

In addition to the quality of sperm parameters, and chromatin integrity that were lower significantly in varicocele group, the expression of TRPV1 protein was also lower in varicocele condition that could be associated with reduced capacitation, acrosome reaction and sperm/oocyte fusion and thereby infertility.

Sperm cryopreservation results in damage to membrane integrity, sperm viability, sperm motility, and DNAstructure. We aimed to evaluate the effect of plasma rich in growth factors (PRGF) on sperm parameters during the freeze-thaw process. In the first phase of this prospective study, after sperm preparation, 10 normozoospermic specimens were cryopreserved by rapid freezing with different concentrations of PRGF including 0, 1, 5, and 10% to find the optimum dose. Sperm motility and viability were assessed in this phase. In the second phase of the study, based on the results of first phase, 25 normal sperm samples were frozen with 1% PRGF. All sperm parameters including motility, viability, acrosome reaction, and DNA integrity were assessed before freezing and after thawing. The rates of progressive and total sperm motility and viability were significantly higher in 1% PRGF compared to control, 5%, and 10% PRGF in the first phase (P<0.05). Supplementation of freezing medium with 1% PRGF could significantly improve all sperm parameters including sperm motility, viability, normal morphology, acrosome integrity, chromatin structure, chromatin integrity, DNA denaturation, and DNA fragmentation in comparison with the control group. It appears that the supplementation of freezing medium with 1% PRGF could protect human sperm parameters during cryopreservation.

This study was designed to determine the effects of pre-ischemic administration of oxytocin (OXT) on neuronal injury and possible molecular mechanisms in a mice model of stroke. In this experimental study, stroke was induced in the mice by middle cerebral artery occlusion (MCAO) for 60 minutes and 24 hours of reperfusion. OXT was given as intranasal daily for 7 consecutive days before ischemic stroke. Neuronal damage, spatial memory, and the expression levels of nuclear factor-kappa B (NF-κB), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9), brain-derived neurotrophic factor (BDNF) and apoptosis were assessed 24 hours after stroke. Pre-ischemic treatment with OXT significantly reduced the infarct size (P<0.01); but did not recover the neurological and spatial memory dysfunction (P>0.05). Moreover, OXT treatment considerably decreased the expressions of NF-κB, TNF-α, IL-1β, and MMP-9 (P<0.001) and enhanced the level of BDNF protein. OXT treatment also significantly downregulated Bax expression and overexpressed Bcl-2 proteins. The finding of this study indicated that administration of OXT before ischemia could limit brain injury by inhibiting MMP-9 expression, apoptosis, inflammatory signaling pathways, and an increase in the BDNF protein level. We suggested that OXT may be potentially useful in the prevention and/or reducing the risk of the cerebral stroke attack, and could be offered as a new prevention option in the clinics.

Bone regeneration is a desired treatment outcome in implant dentistry. The primary goal of the current investigation was to assess the joint effect of low-level laser therapy (LLLT) and leukocyte- and platelet-rich fibrin (PRF) on new bone formation.

During this experiment study, forty bone defects (8 mm in diameter) were generated in the calvaria of ten New-Zealand white rabbits. defects were filled with autogenous bone defined as the control group, autogenous bone with leukocyte- and PRF (PRF group), autogenous bone and low-level diode laser radiation (LLLT group), and autogenous bone with leukocyte- and PRF and low-level laser radiation (LP group). Laser irradiation was done every second day for 2 weeks after surgery. Five rabbits were randomly selected to be sacrificed on postoperative weeks 4 and 8. On one and two-month post-surgery, histological and histomorphometric parameters including bone formation, fibroblast, and osteoblast were assessed.

The histological panel depicted that the ratio of fresh bone formation increased at one-and two-month postsurgery in all treatment groups compared to the control group. The most favorable results were seen in the LP group, followed by the PRF group. Based on the ANOVA test, bone neoformation was statistically significant in the LP group in comparison with the control group (P<0.001). One-month post-surgery, a higher degree of fibroblast was seen in the control group, while the last place was for LP group (118.6 ± 6.9 vs. 24.0 ± 3.2). In the PRF group, the percentage ofbone formation was higher than that in the control group (13.2 ± 2.8 vs. 2.0 ± 1.2), but no significant difference when compared to the LP group (13.2 ± 2.8 vs. 19.0 ±.3.8).

The combined L-PRF and LLLT was more likely to have a positive effect on accelerating bone regeneration and reducing fibrosis.