Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
Novel Variant Identified in the Enhancer Region of Host Transcription Factor, BRN3A, is a Significant Risk Factor for HPV-Induced Uterine Cervix Cancer
88
103
EN
Anand
Prakash
Department of Zoology, Banaras Hindu University, Varanasi, India.
Biswa Pratim
Das Purkayastha
Department of Zoology, Banaras Hindu University, Varanasi, India.
Shikha
Srivastava
Department of Zoology, Banaras Hindu University, Varanasi, India.
Sunanda
Chaturvedi
Homi Bhabha Cancer Hospital & Research Centre, Varanasi, India.
Akhtar
Ali
Centre for Genetic Disorders, Banaras Hindu University, Varanasi, India.
Dau Dayal
Aggarwal
Department of Zoology, Banaras Hindu University, Varanasi, India.
Jagat Kumar
Roy
Department of Zoology, Banaras Hindu University, Varanasi, India.
Among HPV-mediated cervical cancers, cellular factor BRN3A has gained considerable attention due to its role in promoting an anti-apoptotic cellular environment and in facilitating epitheliotropic transformations of the host. The majority of previous studies looked at BRN3A's molecular characteristics; however, the possibility of genetic variations in BRN3A's auto-regulatory region in relation to cervical cancer risk has been underestimated until now. In a retrospective study in the Eastern UP population, India, we detected genetic variations in the cis-regulatory proximal enhancer region located around 5.6 kb upstream of transcription start site of BRN3A. Our analysis of PCR and DNA sequencing confirmed this novel SNP (BRN3A g.60163379A>G) within the auto-regulatory region of BRN3A. As compared to control subjects, cancer cases exhibited a 1.32-fold higher allele frequency (χ2 = 6.315, p = 0.012). In homozygous (GG) but not in heterozygous conditions, odds ratio (OR) analysis suggests a significant association of cancer risk with the SNP (OR = 2.60, p ≤ 0.004). We further confirmed using the functional analysis that this SNP increased the luciferase gene activity in HPV-positive cervical cancer SiHa cells that were exposed to progesterone. As a result of the association of polymorphisms in a non-coding region of an oncogene with increased cancer risks, we are suggesting that this genetic variation in non-coding region can be used in prediction, diagnosis, or predicting the progression of the disease.
Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
Sulforaphane, a Chemopreventive Compound Induces Necrotic Behavior and Inhibits S-phase of Cell Cycle in Human Kidney Cells in vitro
104
116
EN
Guzin
Gokay
Bezmialem Vakif University, Health Sciences Institute, Department of Biotechnology, Istanbul, Turkey.
Beyza
Goncu
Bezmialem Vakif University, Vocational School of Health, Department of Medical Laboratory Techniques, Istanbul, Turkey.
Sezen
Atasoy
Bezmialem Vakif University, Faculty of Pharmacy, Biochemistry, Istanbul, Turkey.
Nur
Ozten Kandas
Bezmialem Vakif University, Faculty of Pharmacy, Pharmaceutical Toxicology, Istanbul.
Aydan
Dag
Bezmialem Vakif University, Faculty of Pharmacy, Pharmaceutical Chemistry, Istanbul.
Sulforaphane (SFN) is an organosulfur product of found isothiocyanates in vegetables. The chemopreventive effects of SFN have revealed that there is a link between excessive consumption of SFN-rich vegetables and cancer formation without possible toxicological consequences. We aimed to evaluate the cellular outcome of SFN from a toxicological perspective, particularly for renal cells including clear cell adenocarcinoma (769-P) and human embryonic renal epithelial (293T) cells. The viability/cytotoxicity experiments were performed with methyl thiazole diphenyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays. IC50-dependent, non-cytotoxic concentrations were used for the determination of cell cycle status and apoptosis by using flow cytometry and western blot. A certain concentration of SFN effectively altered apoptotic/necrotic behavior in 769-P compared to the control group 293T. Cell cycle status remained stable while showing a decreased proliferation profile for 769-P cells. The percentage of the S phase from the cell cycle in 293T cells significantly reduced without affecting proliferation status. The use of SFN as an alternative to traditional treatments might be considered for the battle against renal cell carcinoma but the current findings showed that caution should be applied particularly for renal cells. Our study will provide a basis for future in vivo studies to support traditional cancer therapies.
Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
A Decrease in CD44 on Cell Surfaces (MKN-45 Cell Line) After RELA Knockout Using CRISPR/Cas9
117
126
EN
Saeid
Karimi
Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Sima
Salmani
Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Akram
Alizadeh
Department of Tissue Engineering and Applied Cell Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran.
Leila
Rezakhani
Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Zohreh
Saltanatpour
Pediatric Cell and Gene Therapy Research Center, Gene, Cell & Tissue Research Institute, Tehran University of Medical Sciences, Tehran, Iran.
Sorayya
Ghasemi
Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
The NF-kB signaling pathway was introduced as a key pathway in carcinogenesis that is induced by inflammation in gastrointestinal malignancies. The RelA transcription factor is an important component of this signaling pathway. Furthermore, CD44 is implicated in the tumorigenesis and metastasis of gastric cancer. The aim of this study was to assay the effect of RELA knockout on CD44 expression in MKN45 cells. CRISPR/Cas9 was used to knock out RELA in MKN-45. The median fluorescence intensity (MFI) of CD44 before and after RELA knockout is analyzed in MKN45. The CRISPR/Cas9 vector pSpCas9 (BB)-2A-Puro (PX459) was used for gRNA cloning (two guides). The MKN-45 cell line was co-transfected. The purified co-transfected cells with puromycin were cultured and used for the RELA gene expression assay by real-time PCR. Flow cytometry was used for the analysis of the MFI of CD44+ in MKN45. The results showed that 180 nucleotide sequences between exon 2 and exon 3 of RELA were deleted in MKN45. RELA expression was significantly (P<0.001) decreased after CRISPR/Cas9 knockout. Compared to the control group, the MFI of CD44 in transfected cells was significantly decreased (P <0.001). Knockout of RELA significantly decreased CD44 expression in MKN45 cells. It can be concluded that the NF-kB signaling pathway via RELA is related to CD44 expression and consequently the tumorigenesis of gastric cancer. More studies about this relationship are recommended.
Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
Nrf2 rs6721961 and Oxidative Stress in Preeclampsia: Association with the Risk of Preeclampsia and Early-Onset Preeclampsia
127
136
EN
Fatemeh
Khadir
Department of Clinical Biochemistry, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Zohreh
Rahimi
Department of Clinical Biochemistry, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Azita
Ghanbarpour
Department of Obstetrics and Gynecology, Clinical Research Development Unit of Rouhani Hospital, Babol, Iran.
Asad
Vaisi-Raygani
Department of Clinical Biochemistry, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Preeclampsia as a multifactor hypertensive disorder of pregnancy is associated with enhanced placental oxidative stress. The Keap1-Nrf2 pathway protects cells against oxidative stress. We examined the possible association between the Nrf2 variants in relation to oxidative stress parameters with the risk of preeclampsia. We studied 150 preeclampsia women and 150 women with a normal pregnancy to find the frequency of Nrf2 rs6721961 genotypes using the PCR-RFLP method. Also, an association between the Nrf2 genotypes with the levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) was analyzed. Significantly lower TAC and higher MDA levels were found in preeclampsia patients compared to controls (P<0.0001). For the first time, we report an association between the Nrf2 rs6721961 polymorphism and preeclampsia risk. The present study indicated that the GT genotype and the T allele of the Nrf2 rs6721961 increased the risk of preeclampsia by 2.81 and 2.39 times, respectively. Also, the Nrf2 TT genotype was associated with a 3.9-fold increased risk of early-onset preeclampsia. We detected a positive association between the levels of body mass index, MDA, and the Nrf2 polymorphism with the risk of preeclampsia and a negative correlation between the level of TAC with the preeclampsia risk. Also, an association between the rs6721961 TT genotype with higher serum MDA levels was found. Our study suggests oxidative stress is involved in the pathogenesis of preeclampsia and the Nrf2 rs6721961 polymorphism through alteration in the levels of oxidative stress parameters might increase the risk of preeclampsia and early-onset preeclampsia.
Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
hsa-miR-508-5p as a New Potential Player in Intervertebral Disc Degeneration
137
149
EN
Akram
Gholipour
Cardiogenetic Research Center, Rajaie Cardiovascular Medical and Research Center, Iran University of Medical Sciences, Tehran, Iran.
Mahshid
Malakootian
Cardiogenetic Research Center, Rajaie Cardiovascular Medical and Research Center, Iran University of Medical Sciences, Tehran, Iran.
Maziar
Oveisee
School of Medicine, Bam University of Medical Sciences, Bam, Kerman, Iran.
Intervertebral disc degeneration (IDD) is widely known as the principal cause of low back pain, diminishing patients’ quality of life and imposing a huge economic burden on healthcare systems worldwide. However, the underlying mechanisms of IDD remain to be determined. This study aimed to scrutinize data sets via bioinformatics to identify microRNAs (miRNAs)/genes and pathways associated with IDD. The array profiling of patients with IDD and individuals without IDD was acquired from the Gene Expression Omnibus (GEO) database (viz., GSE19943, GSE63492, and GSE34095). The expression profiles of miRNAs and genes with differential patterns were analyzed using GEO2R. The target genes of the chosen miRNA were then examined, and in silico functional analyses were performed on the signaling pathways and biological processes of the differentially expressed genes. Three human miRNAs were up and downregulated in IDD patients in the examined data sets. Among them, hsa-miR-508-5p had a significant differential expression in the IDD group, and SEC11A, IPO5, FN1, and MRPS10, as the targets of hsa-miR-508-5p, were upregulated in the IDD group. Furthermore, extracellular matrix-receptor interactions, focal adhesion, and actin cytoskeleton regulation were important pathways involved in IDD. Our analysis identified hsa-miR-508-5p as a novel miRNA involved in IDD pathogenies. Our findings not only further confirm the significant role of miRNAs in IDD pathogenesis but also extend the spectrum of the miRNAs and genes involved in IDD. Still, further experimental investigations are needed to confirm our findings.
Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
miR-33-5p Regulates CREB to Induce Morphine State-dependent Memory in Rats: Interaction with ยต Opioid Receptor
150
167
EN
Sadegh
Moradi-Vastegani
Institute for Basic Sciences, Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran.
Behrang
Alani
Department of Applied Cell Sciences, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
Khatereh
Kharazmi
Institute for Basic Sciences, Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran.
Abolfazl
Ardjmand
Institute for Basic Sciences, Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran.
The aim of the present study was to examine the hypothesis that miR-33-5p attenuates morphine state-dependent (StD) memory via the µ opioid receptor by regulating cyclic AMP response element-binding protein (CREB). The effects of post-training morphine and morphine StD memory and their interaction with pre-test naloxone were evaluated using a single-trial inhibitory avoidance paradigm. Then, the hippocampal miR-33-5p gene and pCREB/CREB protein expression profiles were evaluated using quantitative real-time PCR and western blotting, respectively. We found that while post-training morphine and morphine StD memory respectively up- and down-regulate the miR-33-5p expression profile in the hippocampus, the reverse results are true for the expression of pCREB/CREB. Pre-test naloxone antagonized the response. Overall, our findings suggest that the expression levels of miR-33-5p in the hippocampus set the basis for morphine StD memory with low miR-33-5p enabling state dependency. The mechanism is mediated via miR33-5p and CREB signaling with the interaction of the µ opioid receptor. This finding may be used as a potential strategy for ameliorating morphine-induced memory-related disorders.
Babol University of Medical Sciences
International Journal of Molecular and Cellular Medicine (IJMCM)
2251-9637
11
2
2022
11
1
Special Considerations on Insulin Resistance Induced Hyperandrogenemia in Women with Polycystic Ovary Syndrome: A Prominent COVID-19 Risk Factor
168
179
EN
Jamshid
Roozbeh
Shiraz Nephro-Urology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Sahar
Janfeshan
Shiraz Nephro-Urology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Afsoon
Afshari
Shiraz Nephro-Urology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Aida
Doostkam
Shiraz Nephro-Urology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Ramin
Yaghobi
Shiraz Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) infecting mechanism depends on hosting angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2) as essential components and androgens as regulators for inducing the expression of these components. Therefore, hyperandrogenism-related disease such as polycystic ovary syndrome (PCOS) in insulin resistant women in reproductive-age is a high-risk factor for SARS-CoV-2 infection. Here, we describe the signaling pathways that might increase the susceptibility and severity of this new pandemic in PCOS women with insulin resistance (IR). Luteinizing hormone and insulin increase the risk of SARS-CoV-2 infection in these patients via induction of steroidogenic enzymes expression through cAMP-response element binding protein and Forkhead box protein O1 (FOXO1), respectively. TMPRSS2 expression is activated through phosphorylation of FOXO1 in ovaries. In other words, SARS-CoV-2 infection is associated with temporary IR by affecting ACE2 and disturbing β-pancreatic function. Therefore, PCOS, IR, and SARS-CoV-2 infection are three corners of the triangle that have complicated relations, and their association might increase the risk of SARS-CoV-2 infection and severity.