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. 2019 Jul 9;9(18):5227–5245. doi: 10.7150/thno.34273

Figure 5.

Figure 5

PCNAP1 up-regulates PCNA through competing for miR-154. (A) The relative levels of miR-154 were examined by RT-qPCR in clinical HBV-related HCC samples. (B and C) The correlation between the PCNAP1 or PCNA mRNA levels and the miR-154 levels was examined by RT-qPCR in clinical HBV-related HCC samples. (D) Luciferase activities of PCNA mRNA 3′UTR were measured by luciferase reporter gene assays in 293T cells transfected with miRNA control, miRNA-154 or miRNA-154 inhibitors. (E) The expression of miR-154 and PCNA was detected by RT-qPCR and Western blot analysis in HepG2.2.15 cells transfected with miRNA control or miRNA-154. (F) The RNA levels of PCNAP1, miR-154 and PCNA were measured by RT-qPCR analysis in HepG2 cells with the single treatment of PCNAP1 or miR-154 and the co-transfection with PCNAP1 and miR-154. (G) The RNA levels of PCNAP1, miR-154 and PCNA were tested by RT-qPCR analysis in HepG2.2.15 cells with the single treatment of siPCNAP1 or anti-miR-154 and the co-transfection with siPCNAP1 and anti-miR-154. (H) The protein levels of PCNA were detected by Western blot analysis in the indicated cells with different treatments as above. (I) Luciferase activities of PCNA 3′UTR were examined in HepG2.2.15 cells with indicated regiments. Error bars represent means ± SD (n=3). Statistical significant differences are indicated: *P<0.05; **P<0.01; ***P<0.001; NS, no significance; Student's t test.