Fig 1: Rutin can regulate autophagy through the BANCR/miRNA-590-5P/OLR1 axis. (A-C) Rutin treatment inhibited the autophagic activity and OLR1 levels in HepG2/SO and HCCLM3/SO cells. Moreover, up-regulation of BANCR increased the autophagic activity in HepG2/SO and HCCLM3/SO cells. (D-E) After sh-BANCR or si-OLR1 treatment, the autophagic activity was weakened in HepG2/SO and HCCLM3/SO cells. The yellow arrow points to the autophagosome. SO: Sorafenib; BANCR: BRAF-activated non-protein coding RNA; OLR1: OLR1: oxidized low-density lipoprotein receptor 1. The scale bar is 2 um.
Fig 2: LncRNA BANCR acts as a molecular sponge of miRNA-590-5P to sequester miRNA-590-5P away from OLR1 in HCC cells. (A) The miRNA-590-5P level was markedly increased in the sh-BANCR group, but was prominently decreased in the Lv-BANCR group. * Compared with the control group, P<0.05. (B) Venn diagram was used to analyze the target proteins through PicTar, miRanda, and Targetscan online prediction systems. A total of 3 target protein was predicted, namely OLR1, GATA3, and Gab2. (C) Western blot results confirmed that miRNA-590-5P up-regulation suppressed OLR1 level, while this effect was reversed by BANCR up-regulation in HepG2 and HCCLM3 cells. (D) The effects of miRNA-590-5P overexpression on luciferase activity of WT-BANCR and Mu-BANCR reporters or OLR1 3'UTR-WT and OLR1 3'UTR-Mu reporters were determined in HepG2 cell. * Compared with the miR-control group, P<0.05. (E) RNA pull-down assay was used, and we found that BANCR and OLR1 could specifically interact with miR-590-5p (qRT-PCR was used). NS was the biotin-labeled non-sense RNA with a similar length to miR-590-5p, NC was the miR-590-5p without biotin label. One-way ANOVA was used. * Compared with the biotin-miR-590-5p group, P<0.05. (F) Compared with chemosensitive tissues and adjacent tissues, the levels of OLR1 protein in chemoresistance tissues were significantly higher. One-way ANOVA was used. (G-H) miRNA-590-5P level dramatically decreased, while the OLR1 level prominently increased in HCC tissues (qRT-PCR was used). * Compared with adjacent tissues, P<0.05. (I-K) BANCR was positively correlated with OLR1, and negatively correlated with miRNA-590-5P in HCC tissues (Pearson correlation analysis was used). SO: Sorafenib; lncRNA: long non-coding RNA; BANCR: BRAF-activated non-protein coding RNA; OLR1: OLR1: oxidized low-density lipoprotein receptor 1. The scale bar is 100 um. Data were expressed as mean ± SEM.
Fig 3: Rutin can enhance the efficacy of SO in a xenograft model of HCC in nude mice. (A-B) Intraperitoneal injection of rutin and SO alone for 14 days inhibited tumor growth. One-way ANOVA was used. * Compared with the control group, P<0.05; # Compared with the Rutin group, P<0.05; $ Compared with the Sorafenib group, P<0.05. (C) Intraperitoneal injection of rutin and SO alone for 14 days inhibited BANCR expression (qRT-PCR was used). One-way ANOVA was used. * Compared with the control group, P<0.05; # Compared with the Rutin group, P<0.05; $ Compared with the Sorafenib group, P<0.05. (D) Intraperitoneal injection of rutin and SO alone for 14 days promoted miRNA-590-5P expression (qRT-PCR was used). One-way ANOVA was used. * Compared with the control group, P<0.05; # Compared with the Rutin group, P<0.05; $ Compared with the Sorafenib group, P<0.05. (E) Intraperitoneal injection of rutin and SO alone for 14 days inhibited OLR1 expression. SO: Sorafenib; BANCR: BRAF-activated non-protein coding RNA; OLR1: OLR1: oxidized low-density lipoprotein receptor 1.
Supplier Page from Abcam for Mouse LOX1 ELISA Kit (OLR1)