Cell Journal (Yakhteh)

285-293

Authors: Shimin Wu; Juan Chen; Ying Liang; Qian Luo; Yaoyao Tong; Ling Xie

Objectives: 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. Materials and Methods: 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. Results: 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. Conclusion: 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.

302-308

Authors: Mohammad Hossein Derakhshan Nazari; Rana Askari Dastjerdi; Parnian Ghaedi Talkhouncheh; Ahmad Bereimipour; Hamidreza Mollasalehi; Amir Ali Mahshad; Ali Razi; Mohammad Hossein Nazari; Amin Ebrahimi Sadrabadi; Sara Taleahmad

Objectives: 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. Materials and Methods: In this bioinformatics study, we compared the expression profile of 7 minor stages of NSCLC adenocarcinoma, 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. Results: Primary analysis showed that BMP4 was downregulated through the tumor progression to the stage IB and GPX2 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-� 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. Conclusion: 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.

316-322

Authors: Hadis Abdolahzadeh; Parvaneh Mohammadi; Mahshid Ghasemi; Seyed Ahmad Mousavi; Amir Bajouri; Leila Ataei-Fashtami; Mehdi Totonchi; Mohammad Rezvani; Nasser Aghdami; Saeed Shafieyan

Objective: 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. Materials and Methods: 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. Results: 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-regulated genes. Upregulated genes were involved in processes, such as Fatty Acid Omega Oxidation. Down-regulated genes were 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. Conclusion: 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).

330-336

Authors: Jafar Mirzaei; Mansoureh Movahedin; Iman Halvaei

Objectives: Sperm cryopreservation results in damage to membrane integrity, sperm viability, sperm motility, and DNA structure. We aimed to evaluate the effect of plasma rich in growth factors (PRGF) on sperm parameters during the freeze-thaw process. Materials and Methods: 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. Results: 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. Conclusion: It appears that the supplementation of freezing medium with 1% PRGF could protect human sperm parameters during cryopreservation.

346-352

Authors: Fereshteh Shanei; Ahad Khoshzaban; Ferial Taleghani; Maryam Tehranchi; Mohammad Hossein Ta

Objectives: 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. Materials and Methods: 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. Results: 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 of bone 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). Conclusion: The combined L-PRF and LLLT was more likely to have a positive effect on accelerating bone regeneration and reducing fibrosis.