Objective: To determine miRNA-34a regulated cell senescence indirectly through targeting silent mating-type information regulation 2 homologue 1 (SIRT1) in vitro experiment.
Methods: A constructed pre-miRNA -34a expression vector and a miRNA-1792 expression vector (not directly against any gene) were transfected into HEK293 and HUVEC cell lines respectively. The expression levels of SIRT1 in each cell groups were detected by RT-PCR and Western blot. The HUVEC cells were divided into different group: transfected with pre-miRNA-34a expression vector (HUVEC-pre-miRNA-34a), transfected with miRNA-1792 expression vector (HUVEC-pre-miRNA-1792), treated HUVEC cell with SIRT1 activator resveratrol (final concentration 1 micromol/L, treatment for 2 h)(HUVEC-Res), and HUVEC cells without any treatment as the control. Comet assay was applied to detect the oxidative damage of above-mentioned cells after H2O2 treatment for 2 h, and beta-galactosidase (SA-beta-gal) staining was used to detect the senescence of them in different time points after doxorubicin treatment for 2 h.
Results: Pre-miRNA-34a expression vector was constructed successfully by sequencing confirmation. RT-PCR and Western blot indicated that the overexpression of miRNA-34a down regulated mRNA and protein level of SIRT1 in HEK293-miRNA-34a and HUVEC-miRNA-34a cell groups (P < 0.001). Comet assay revealed that HUVEC-miRNA-34a cell group was the most sensitive to H2O2 treatment, and the DNA damage of HUVEC-Res cell group was the most minor. HUVEC-miRNA-34a cell group displayed higher frequency of SA-beta-gal staining than that of other cell groups.
Conclusion: miRNA-34a regulated cell senescence indirectly through targeting SIRT1.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Expression and purification of recombinant tilapia lake virus segment 4 protein and its in-vitro biological activity for potential use in vaccine development.
Sci Rep
December 2024
Department of Biotechnology, Faculty of Agro-industry, Kasetsart University, Bangkok, 10900, Thailand.
Tilapia lake virus (TiLV) disease is highly contagious and causes substantial mortality in tilapia. Currently, no effective treatments or commercial vaccines are available to prevent TiLV infection. In this study, TiLV segment 4 (S4) was cloned into the pET28a(+)vector and transformed into Escherichia coli BL21(DE3).
View Article and Find Full Text PDFDecoding of pain during heel lancing in human neonates with EEG signal and machine learning approach.
Sci Rep
December 2024
Neuroscience & Neuroengineering Research Lab, Biomedical Engineering Department, School of Electrical Engineering, Iran University of Science & Technology (IUST), Narmak, Tehran, Iran.
Currently, pain assessment using electroencephalogram signals and machine learning methods in clinical studies is of great importance, especially for those who cannot express their pain. Since newborns are among the high-risk group and always experience pain at the beginning of birth, in this research, the severity of newborns has been investigated and evaluated. Other studies related to the annoyance of newborns have used the EEG signal of newborns alone; therefore, in this study, the intensity of newborn pain was measured using the electroencephalogram signal of 107 infants who were stimulated by the heel lance in three levels: no pain, low pain and moderate pain were recorded as a single trial and evaluated.
View Article and Find Full Text PDFChuanxiong Rhizoma regulates ferroptosis and the immune microenvironment in ischemic stroke through the JAK-STAT3 pathway.
Sci Rep
December 2024
Acupuncture and Moxibustion College, Liaoning University of Traditional Chinese Medicine, Shenyang, 110847, China.
Ferroptosis is linked to various pathological conditions; however, the specific targets and mechanisms through which traditional Chinese medicine influences ischemic stroke (IS)-induced ferroptosis remain poorly understood. In this study, data from the Gene Expression Omnibus and disease target databases (OMIM, GeneCards, DisGeNet, TTD, and DrugBank) were integrated with ferroptosis-related gene datasets. To identify key molecular targets of Chuanxiong Rhizoma (CX), drug ingredient databases, including PubChem and TCMBank, were employed to map CX-related targets (CX-DEGs-FRG and CX-IS-FRG).
View Article and Find Full Text PDFEvidence for the replication of a plant rhabdovirus in its arthropod mite vector.
Virus Res
December 2024
Institute of Plant Science and Resources (IPSR), Okayama University, Kurashiki 710-0046, Japan.
Transmission of plant viruses that replicate in the insect vector is known as persistent-propagative manner. However, it remains unclear whether such virus-vector relationships also occur between plant viruses and other biological vectors such as arthropod mites. In this study, we investigated the possible replication of orchid fleck virus (OFV), a segmented plant rhabdovirus, within its mite vector (Brevipalpus californicus s.
View Article and Find Full Text PDFDesign and Cytotoxicity Evaluation of a Cancer-targeting Immunotoxin Based on a Camelid Nanobody-PE Fusion Protein.
Iran J Immunol
December 2024
Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Background: Developing effective targeted treatment approaches to overcome drug resistance remains a crucial goal in cancer research. Immunotoxins have dual functionality in cancer detection and targeted therapy.
Objective: This study aimed to engineer a recombinant chimeric fusion protein by combining a nanobody-targeting domain with an exotoxin effector domain.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!