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INTRODUCTION: The present study was to investigate the clinical role of miR-330-3p in acute cerebral infarction (ACI), including its diagnostic and prognostic potential. Preliminary exploration of its target genes was archived by bioinformatics analysis. METHODS: miR-330-3p in plasma of the patients with ACI and controls were quantified by real-time quantitative PCR. The one-month prognosis of the ACI patients was evaluated by the Glasgow outcome scale (GOS). The correlation between the plasma levels of miR-330-3p and the GOS scores was tested by Pearson correlation analysis. The receiver operating characteristic (ROC) curves were established based on the expression level of miR-330-3p in different groups. The miR-330-3p-targeting genes were analyzed using Venn diagram, protein-protein interaction network, and Gene Ontology enrichment analysis. RESULTS: MiR-330-3p was significantly increased in the plasma of ACI patients compared with that in healthy controls, and ROC curve revealed its diagnostic value for ACI. miR-330-3p was significantly increased in the plasma of patients with poor one-month prognosis compared with those with good one-month prognosis. MiR-330-3p expression was negatively correlated with GOS score, suggesting its potential to predict the one-month prognosis for ACI. One-year survival analysis revealed surviving patients had lower levels of miR-330-3p than the deceased. miR-330-3p was proven to predict the death of patients with ACI. The miR-330-3p-targeting genes were associated with synapse-related Gene Ontology terms. CONCLUSION: MiR-330-3p was upregulated in the plasma of patients with ACI, making it a promising diagnostic and prognostic marker for patients with ACI. MiR-330-3p could facilitate synaptic plasticity following cerebral infarction.
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OBJECTIVE: Fibroblast activation protein (FAP) has been widely studied and exploited for its clinical applications. One of the difficulties in interpreting reports of FAP-targeted theranostics is due to the lack of accurate controls, making the results less specific and less confirmative. This study aimed to establish a pair of cell lines, in which one highly expresses FAP (HT1080-hFAP) and the other has no detectable FAP (HT1080-vec) as control, to accurately evaluate the specificity of the FAP-targeted theranostics in vitro and in vivo. METHODS: The cell lines of the experimental group (HT1080-hFAP) and no-load group (HT1080-vec) were obtained by molecular construction of the recombinant plasmid pIRES-hFAP. The expression of hFAP in HT1080 cells was detected by PCR, Western blotting and flow cytometry. CCK-8, Matrigel transwell invasion assay, scratch test, flow cytometry and immunofluorescence were used to verify the physiological function of FAP. The activities of human dipeptidyl peptidase (DPP) and human endopeptidase (EP) were detected by ELISA in HT1080-hFAP cells. PET imaging was performed in bilateral tumor-bearing nude mice models to evaluate the specificity of FAP. RESULTS: RT-PCR and Western blotting demonstrated the mRNA and protein expression of hFAP in HT1080-hFAP cells but not in HT1080-vec cells. Flow cytometry confirmed that nearly 95% of the HT1080-hFAP cells were FAP positive. The engineered hFAP on HT1080 cells had its ability to retain enzymatic activities and a variety of biological functions, including internalization, proliferation-, migration-, and invasion-promoting activities. The HT1080-hFAP xenografted tumors in nude mice bound and took up 68GA-FAPI-04 with superior selectivity. High image contrast and tumor-organ ratio were obtained by PET imaging. The HT1080-hFAP tumor retained the radiotracer for at least 60 min. CONCLUSION: This pair of HT1080 cell lines was successfully established, making it feasible for accurate evaluation and visualization of therapeutic and diagnostic agents targeting the hFAP.