Fig. 4

Midazolam negatively regulated HOOK3 via upregulating miR-194-5p. A The targeting sites between miR-194-5p and 3’UTR of HOOK3 were predicted by our bioinformatics analysis, and the bindings sites in HOOK3 were mutated. B, C The dual-luciferase reporter gene system assay was conducted to validate the targeting sites in miR-194-5p and HOOK3. Upregulation of miR-194-5p suppressed HOOK3 expressions at both D mRNA and E, F protein levels, as determined by Real-Time qPCR and Western Blot analysis. Real-Time qPCR analysis elucidated that HOOK3 mRNA tended to be enriched in CR-NSCLC G cells and H clinical tissues. I MiR-194-5p and HOOK3 mRNA were negatively correlated in the clinical NSCLC tissues, as determined by Pearson correlation analysis. J–L Knock-down of miR-194-5p rescued HOOK3 expressions in the cisplatin and midazolam co-treated CR-NSCLC cells. Individual experiment contained at least 3 repetitions, and *P < 0.05, **P < 0.01, and ***P < 0.001 was considered as statistical significance