Fig 1: lncRNA F11-AS1 positively regulates NR1I3 expression via competitively binding to miR-211-5p. A, The complementary binding sequences between miR-211-5p and NR1I3 3’UTR predicted by bioinformatics analysis. B, The NR1I3 expression in HBV+/HBV- HCC tissues and corresponding adjacent tissues measured by RT-qPCR and immunohistochemistry; * P < .05 vs adjacent tissues or HBV- HCC tissues; HBV+ HCC tissues = 45, HBV- HCC tissues = 28. C, The mRNA and protein expression of NR1I3 in HepG2 and HepG2.2.15 cells evaluated by RT-qPCR and Western blot analysis; * P < .05 vs the HepG2 cells. D, The mRNA and protein expression of NR1I3 in Huh7 and HepG2 cells following transfection with pHBV1.3 or pUC18 assessed by RT-qPCR and Western blot analysis; * P < .05 vs the Huh7 or HepG2 cells transfected with pUC18. E, The relationship between miR-211-5p and NR1I3 confirmed by dual-luciferase reporter assay; * P < .05 vs the HEK-293T transfected with NC-mimic. F, The binding ability of miR-211-5p to NR1I3 detected by RNA pull-down; *P < .05 vs the HepG2.2.15 cells incubated with Bio-NC. G, the mRNA expression of NR1I3 in the transfected HepG2.2.15 cells measured by RT-qPCR;* P < .05 vs the HepG2.2.15 cells transfected with NC-mimic; # P < .05 vs the HepG2.2.15 cells delivered with miR-211-5p inhibitor. H, the protein expression of NR1I3 in the transfected HepG2.2.15 cell determined by Western blot analysis; # P < .05 vs the HepG2.2.15 cells transfected with miR-211-5p inhibitor. I, the protein expression of NR1I3 detected by immunocytochemistry. J, the binding status of miR-211-5p and NR1I3 detected by Northern blot analysis. K, Pearson's correlation analysis of lncRNA F11-AS1, miR-211-5p and NR1I3 in patients with HBV+-related HCC. Data were measurement data and expressed by mean ± standard deviation. Comparison of data between the two groups was analysed by unpaired t test, and that among multiple groups was examined by one-way analysis of variance followed by Tukey's post hoc test. Pearson's correlation coefficient was used for correlation analysis
Fig 2: lncRNA F11-AS1 up-regulates the expression of NR1I3 via miR-211-5p to hinder the progression of HBV-related HCC. In HBV-related HCC, HBV-encoded HBx protein inhibited the expression of lncRNA F11-AS1. lncRNA F11-AS1 could up-regulate NR1I3 via binding to miR-211-5p, whereas down-regulation of lncRNA F11-AS1 caused by HBx protein weakened its ability to bind to miR-211-5p, which could target to decrease NR1I3 expression. As a result, the proliferation, migration and invasion of HBV-related HCC cells were enhanced, yet cell apoptosis was attenuated. Importantly, overexpression of lncRNA F11-AS1 could enhance the NR1I3 expression by acting as a ceRNA of miR-211-5p, ultimately impeding the progression of HBV+ HCC
Fig 3: lncRNA F11‐AS1/miR‐211‐5p/NR1I3 axis regulates the proliferation, apoptosis, migration and invasion of HepG2.2.15 cells. A, The mRNA expression of Bax, Bcl‐2 and PCNA in HepG2.2.15 cells after transduction determined by RT‐qPCR. B, The protein expression of Bax, Bcl‐2 and PCNA in HepG2.2.15 cells after transduction measured by Western blot analysis. C, The proliferative ability of HepG2.2.15 cells after transduction assessed by CCK‐8 assay. D, the apoptosis rate of HepG2.2.15 cells after transduction evaluated by TUNEL staining. E, The migration ability of HepG2.2.15 cells after transduction assessed by scratch test (×40). F, The invasion ability of HepG2.2.15 cells after transduction determined by Transwell assay (×200). Data were measurement data and expressed by mean ± standard deviation. * P < .05 vs the HepG2.2.15 cells delivered with oe‐NC. Comparison of data among multiple groups was examined by one‐way or two‐way analysis of variance followed by Tukey's post hoc test
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