ABSTRACT
Atherosclerosis is a fatal disorder that is fundamental to various cardiovascular diseases and severely threatens people's health worldwide. Several studies have demonstrated the role of circular RNAs (circRNAs) in the pathogenesis of atherosclerosis. circUSP36 acts as a key modulator in the progression of atherosclerosis, but the molecular mechanism underlying this role is as yet unclear. This study aimed to elucidate the mechanism by which circUSP36 exerts its function in an in vitro cell model of endothelial cell dysfunction, which is one of pathological features of atherosclerosis. The circRNA traits of circUSP36 were confirmed, and we observed high expression of circUSP36 in endothelial cells exposed to oxidized low-density lipoprotein (ox-LDL). Functional assays revealed that overexpression of circUSP36 suppressed proliferation and migration of ox-LDL-treated endothelial cells. In terms of its mechanism, circUSP36 adsorbed miR-637 by acting as an miRNA sponge. Moreover, enhanced expression of miR-637 abated the impact of circUSP36 on ox-LDL-treated endothelial cell dysregulation. Subsequently, the targeting relationship between miR-637 and WNT4 was predicted using bioinformatics tools and was confirmed via luciferase reporter and RNA pull-down assays. Notably, depletion of WNT4 rescued circUSP36-mediated inhibition of endothelial cell proliferation and migration. In conclusion, circUSP36 regulated WNT4 to aggravate endothelial cell injury caused by ox-LDL by competitively binding to miR-637; this finding indicates circUSP36 to be a promising biomarker for the diagnosis and therapy of atherosclerosis.
Subject(s)
Atherosclerosis , Endothelial Cells/metabolism , MicroRNAs/genetics , RNA, Circular/genetics , Wnt4 Protein/genetics , Atherosclerosis/genetics , Atherosclerosis/metabolism , Atherosclerosis/physiopathology , Cells, Cultured , Humans , MicroRNAs/metabolism , RNA, Circular/metabolism , Wnt4 Protein/metabolismABSTRACT
Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher's attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with Kd 56±7.3nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.
Subject(s)
Aptamers, Nucleotide/genetics , Aptamers, Nucleotide/metabolism , ErbB Receptors/genetics , ErbB Receptors/metabolism , Neoplasms, Experimental/physiopathology , Aptamers, Nucleotide/chemistry , Base Sequence , Binding Sites , Cell Line, Tumor , Humans , Jurkat Cells , Molecular Imaging/methods , Molecular Sequence Data , Molecular Targeted Therapy , Neoplasms, Experimental/pathology , Protein Binding , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
OBJECTIVE: To evaluate the molluscicidal effect of a novel nano-silver biological molluscicide in the field and its toxicity to other aquatic organisms. METHODS: A marshland of the Qiupu River was selected as the study site. Four 1 m3 pools were excavated. Four groups, including 300 g/m3 and 600 g/m3 nano-silver biological molluscicide groups, a 2 g/m3 50% niclosamide group, and a natural water (control) group were assigned to the four pools which had 100 Oncomelania hupensis snails and 100 crucian seedlings. The molluscicidal effects and toxicity to the aquatic organisms were observed. RESULTS: The snail death rates were 100% and crucian seedling death rates were 6% in the 600 g/m3 nano-silver biological molluscicide group and 300 g/m3 nano-silver biological molluscicide group in 7 days and 14 days. The snail death rate was 100% in the niclosamide group in 7 days, and the crucian seedling death rate was 100% in the niclosamide group in 3 days. The snail death rates were 7% and 14% in the control group in 7 and 14 days respectively, and the crucian seedling death rate was 22% in the control group. CONCLUSIONS: The novel nano-silver biological molluscicide possesses an excellent molluscicidal capability and non-toxicity to the other aquatic organisms. Therefore, it may become a new, efficient and environment-friendly bio-molluscicide for extensive application.
