Figure 5

PINK1-AS targets miR-34a-5p to exert its tumor-promoting effects in HCC. (a) Analysis of the miRcode (http://www.mircode.org/) website shows that PINK1-AS could bind miR-34a-5p. (b) Relative levels of PINK1-AS mRNA in HCC tissues (n = 20). (c) The expression of PINK1-AS in hepatic epithelial cell and HCC cells was detected by RT-qPCR. (d) Kaplan–Meier survival curves showing the effect of PINK1-AS on overall survival. (e) Expression level of PINK1-AS was negative correlated with miR-34a-5p expression level (P < 0.001). (f,g) The expression level of miR-34a-5p was detected in Huh-7 cells and HepG2 cells after transfection of si-PINK1-AS by RT-qPCR. (h,i) The expression level of PINK1-AS was detected in Huh-7 cells and HepG2 cells after transfection of inhibitor-miR-34a-5p by RT-qPCR. (j) Putative binding sequence of miR-34a-5p in the PINK1-AS. (k) Luciferase reporters containing WT and MUT PINK1-AS transcript was co-transfected with miR-34a-5p mimics or miR-control in 293 cells. Luciferase activity was determined using luciferase reporter system. (l) Luciferase reporters containing PINK1-AS-Luc-WT were co-transfected with or without miR-34a-5p and PINK1-AS, Luciferase activity was determined using dual luciferase reporter system. (m) Expression level of PINK1-AS was positively correlated with ALDOA expression level (P < 0.001). (n) Relative luciferase activity of ALDOA 3′UTR was determined after transfection with miR-34a-5p mimics, miR-34a-5p inhibitor or PINK1-AS plasmid. (o,p) Down-regulation of ALDOA levels in PINK1-AS-silenced Huh-7 cells and HepG2 cells was detected by WB. Statistical data are shown. *P < 0.05; **P < 0.01; ***P < 0.001.