Identification and validation of potential key long noncoding RNAs in sorafenib-resistant hepatocellular carcinoma cells

Figures

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  Figure 1. Resistance of HCC cell lines to sorafenib. (A) The effects of Huh7 cells and sorafenib-resistant Huh7 cells to sorafenib were assessed by CCK8 assay. (B) The effects of HepG2 cells and sorafenib-resistant HepG2 cells to sorafenib were assessed by CCK8 assay. Data are shown as mean ± SEM of three independent experiments.

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  Figure 2. Screening of differentially expressed lncRNAs. (A) The number of differentially expressed lncRNAs in sorafenib-resistant Huh7 and HepG2 cells compared to the parental cells (blue, down-regulated; red, up-regulated). (B) Venn diagrams of differentially expressed lncRNAs in sorafenib-resistant Huh7 and HepG2 cells compared to the parental cells. (C) Volcano plots of differentially expressed lncRNAs between Huh7 and Huh7-S cells. (D) Volcano plots of differentially expressed lncRNAs between HepG2 and HepG2-S cells.

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  Figure 3. The expression profiles of lncRNAs. Heat map was performed to demonstrate the hierarchical clustering of lncRNA expression in sorafenib-resistant Huh7 and HepG2 cells compared to the parental cells. The color depth indicated the elevated (Red) and decreased (Blue) expression level across all samples.

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  Figure 4. Prediction and functional classification of target genes. (A) GO enrichment analysis of significantly differential expressed lncRNAs between Huh7 and Huh7-S cells. (B) GO enrichment analysis of significantly differential expressed lncRNAs between HepG2 and HepG2-S cells. (C) KEGG pathway analysis of significantly differential expressed lncRNAs between Huh7 and Huh7-S cells. (D) KEGG pathway analysis of significantly differential expressed lncRNAs between HepG2 and HepG2-S cells.

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  Figure 5. Validation for lncRNAs expression. (A) The expression level of TCONS_00284048 was validated using RT-qPCR in sorafenib-resistant Huh7 cells. (B) The expression level of TCONS_00284048 was validated in sorafenib-resistant HepG2 cells. (C) The expression level of TCONS_00006019 was validated in sorafenib-resistant Huh7 cells. (D) The expression level of TCONS_00006019 was validated in sorafenib-resistant HepG2 cells. p < 0.0001 compared to parental controls. Data are shown as mean ± SEM of three independent experiments.

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  Figure 6. Function of differentially expressed lncRNAs. (A) The expression levels of TCONS_00284048 as silenced using siRNA in both sorafenib-resistant Huh7 and HepG2 cells. (p = 0.0003 and p < 0.0001 respectively). (B) The expression levels of TCONS_00006019 as silenced using siRNA in both sorafenib-resistant Huh7 and HepG2 cells (p < 0.0001). (C) IC50 values were significantly lower in sorafenib resistant Huh7 cells after knockdown TCONS_00284048 or TCONS_00006019, compared to the siNC Huh7-S cells (p = 0.0002 and p = 0.0001 respectively). (D) IC50 values were significantly lower in sorafenib resistant HepG2 cells after knockdown TCONS_00284048 or TCONS_00006019, compared to the siNC HepG2-S cells (p = 0.0004 and p = 0.0009 respectively). Data are shown as mean ± SEM of three independent experiments.