Objective To investigate the effect of microRNA-584-5p (miR-584-5p) on the biological behavior (proliferation, migration and invasion) of breast cancer cells and its mechanism. Methods Human normal breast epithelial cells MCF10A and breast cancer cells MDA-MB-231, SK-BR-3 and MCF-7 were selected; take MCF-7 cells in logarithmic growth phase, transfect them with LipofectamineTM 2000 transfection kit, and divide them into seven groups: blank group (untransfected MCF-7 cells), mimic-negative control (mimic-NC) group (transfected mimic-NC), miR-584-5p mimic group (transfected miR-584-5p mimic), pcDNA group [transfected with overexpression of matrix metalloproteinase-14 (MMP-14) pcDNA3.1 plasmid negative control (pcDNA3.1)], MMP-14 group [transfected with overexpression of MMP-14 pcDNA3.1 plasmid (pcDNA3.1-MMP-14)], mimic-NC+MMP-14 group (co-transfected with mimic NC and pcDNA3.1-MMP-14), and miR-584-5p mimic+MMP-14 group (co-transfected with miR-584-5p mimic and pcDNA3.1-MMP-14). The mRNA expression levels of miR-584-5p in MCF10A, MDA-MB-231, SK-BR-3 and MCF-7 cells and the expression levels of miR-584-5p and MMP-14 mRNA of MCF-7 cell in each group were detected by fluorescence quantitative PCR. The protein expressions of MMP-14 of MCF-7 cell in each group were detected by Western blotting. The proliferation, migration and invasion of MCF-7 cell in each group were detected by cell counting kit - 8 (CCK-8), scratch test and Transwell test. The targeting relationship between miR-584-5p and MMP-14 was detected by double luciferase reporter gene assay. Results Compared with the human normal mammary epithelial cells MCF10A, the expression levels of miR-584-5p in breast cancer cells MDA-MB-231, SK-BR-3 and MCF-7 were decreased (P<0.05), and the expression level of miR-584-5p in MCF-7 cells was the lowest. Compared with the blank group and the mimic-NC group, the expression level of miR-584-5p of MCF-7 cells in the miR-584-5p mimic group was increased, and the expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number were decreased or reduced (P<0.05). Compared with the blank group and the pcDNA group, the expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number of MCF-7 cells in the MMP-14 group were increased (P<0.05). Compared with the MMP-14 group and the mimic-NC+MMP-14 group, the expression level of miR-584-5p of MCF-7 cells in the miR-584-5p mimic+MMP-14 group was increased, the expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number were decreased or reduced (P<0.05). The expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number of MCF-7 cells in the miR-584-5p mimic+MMP-14 group were higher or morer than those in the miR-584-5p mimic group (P<0.05). The results of double luciferase reporter gene test showed that miR-584-5p could targeted action on the MMP-14 promoter region. Conclusions MiR-584-5p can targetable regulate the expression of MMP-14. Overexpression of miR-584-5p inhibits the proliferation, migration and invasion of breast cancer cells by down-regulating MMP-14.
ObjectiveTo verify the reliability of Anticlot Assistant, a patient self-management system for warfarin therapy assisted by artificial intelligence.MethodsIt was a single-center, prospective cohort study. The eligible 34 participants were recruited consecutively between November 29, 2017 to September 27, 2018 and managed by warfarin therapy via Anticlot Assistant. The recommendations of Anticlot Assistant were examined and verified by the doctors to ensure the security. Medical records were exported from the the background management system. An univariate analysis compared the outcomes between accepted and overridden records and a logistic regression model was built to determine independent predictors of the outcomes. The research team analyzed 153 medical records, which were from 18 participants and were input by 19 doctors. There were 97 records with doctor accepting the suggestion and 56 records with doctor rejecting the suggestion .ResultsWhen the doctors accepted the recommendations, the percentage of the next-test international normalized ratio (INR) in the therapeutic range was higher (64.95% vs. 44.64%, RR=2.298, 95%CI 1.173 to 4.499, P=0.014). The logistic regression analysis revealed that accepting the recommendations was an independent predictor for the next-test INR being in the therapeutic range after controlling potentially confounding factors (OR=2.446, 95%CI 1.103 to 5.423, P=0.028).ConclusionThe algorithm of Anticlot Assistant is reasonable and reliable.