Long Noncoding RNA XIST Promotes Resistance to Lenvatinib in Hepatocellular Carcinoma Cells via Epigenetic Inhibition of NOD2

Figure 3

lncXIST inhibits the expression of NOD2 by binding to EZH2. (a) Heat map of differentially expressed genes in HepG2/R cells after transfection of lncXIST or control siRNAs. (b) Top significantly affected pathways in HepG2/R cells after inhibition of lncXIST based on KEGG pathway enrichment analysis. (c) Seven representative gene mRNA levels in HepG2/R cells after inhibition of lncXIST. (d) HepG2/R, HepG2, and Huh7 cells were transfected as indicated, and the protein levels of NOD2 and GAPDH were measured by western blots. (e) Subcellular distribution of lncXIST was assayed. (f) RIP assay was conducted in HepG2/R cells to show lncXIST coimmunoprecipitation with EZH2, HuR, HMGB1, and Ago2. (g) mRNA levels of EZH2 were measured by RT-PCR. (h) Protein levels of EZH2 were measured by western blots. (i) HepG2/R and Huh7 cells were transfected siRNAs against EZH2 or control siRNA for 24 h, and protein levels of EZH2 were measured. (j) HepG2/R and Huh7 cells were transfected siRNAs against EZH2 or control siRNA for 24 h, then cells were treated with or without lenvatinib (10 μM) for another 24 h, and cell death was measured. (k) Cells were treated as described above, and cellular viabilities were measured. (l) HepG2/R and Huh7 cells were transfected with siRNAs against EZH2 or control siRNA for 24 h, and EZH2 and NOD2 mRNA levels were measured. (m) HepG2/R and Huh7 cells were transfected with siRNAs against EZH2 or control siRNA for 24 h, and indicated proteins were measured by western blots. (n) ChIP-qPCR assay showing EZH2 binding with the promoter region of NOD2 can be attenuated by knockdown of lncXIST. The data was presented as . and .

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