Upregulated lncRNA PRNT promotes progression and oxaliplatin resistance of colorectal cancer cells by regulating HIPK2 transcription

Figure 1 Differential expression analysis of oxaliplatin resistance-related genes in the Gene Expression Omnibus database. A: Volcano plot of the oxaliplatin resistance-related genes showing differential expression between oxaliplatin-sensitive and oxaliplatin-resistant groups; B: Heatmap of oxaliplatin resistance-related genes; C: Protein-protein interaction of oxaliplatin resistance-related genes; D: Hub genes of the oxaliplatin resistance-related genes; E: Heatmap of the correlation between oxaliplatin resistance-related genes and long noncoding RNAs; F: Scatter plot of the correlation between prion protein testis specific (PRNT) and homeodomain interacting protein kinase 2 (HIPK2); G: Venn plot of PRNT-related TFs and HIPK2-related TFs; H: Canonical ZNF184 binding motif via Joint Analysis of the Structural Parameters of Analytical Regulation (JASPAR); I: The binding sites of HIPK2 and ZNF184 predicted via JASPAR. PRNT: Prion protein testis specific; ZNF184: Zinc finger protein 184; HIPK2: Homeodomain interacting protein kinase 2.

Figure 2 Correlation analysis of the prion protein testis specific/zinc finger protein 184/homeodomain interacting protein kinase 2 regulatory axis. A: Box diagram of prion protein testis specific (PRNT)/zinc finger protein 184 (ZNF184)/homeodomain interacting protein kinase 2 (HIPK2) in Gene Expression Omnibus datasets; B: Comparative Toxicogenomics database analysis of PRNT/ZNF184/HIPK2; C: Functional enrichment analysis of PRNT via gene set enrichment analysis. PRNT: Prion protein testis specific; ZNF184: Zinc finger protein 184; HIPK2: Homeodomain interacting protein kinase 2.

Figure 3 The expression of prion protein testis specific in cell lines. A: The expression of prion protein testis specific (PRNT) in HCT116, HCT116/L, LoVo and LoVo/L cell lines; B: Nuclear cytoplasmic distribution of PRNT in the HCT116/L cell line; C: Validation of PRNT overexpression and knockdown in the HCT116/L cell line; D: Validation of PRNT overexpression and knockdown in the LoVo/L cell line. ^aP < 0.05; ^cP < 0.001. PRNT: Prion protein testis specific; NC: Negative control.

Figure 4 The results of MTT in prion protein testis specific overexpression and knockdown HCT116/L cells (A) and LoVo/L cells (B). ^cP < 0.001. PRNT: Prion protein testis specific.

Figure 5 The results of the wound healing scratch assay in prion protein testis specific overexpression and knockdown HCT116/L cells (A) and LoVo/L cells (B). ^aP < 0.05; ^bP < 0.01; ^cP < 0.001. PRNT: Prion protein testis specific; NC: Negative control.

Figure 6 The results of invasion assays in prion protein testis specific overexpression and knockdown HCT116/L cells (A) and LoVo/L cells (B). ^aP < 0.05; ^bP < 0.01; ^cP < 0.001. PRNT: Prion protein testis specific; NC: Negative control.

Figure 7 The expression of homeodomain interacting protein kinase 2 and zinc finger protein 184 in prion protein testis specific overexpression and knockdown HCT116/L cells (A) and LoVo/L cells (B) by westernblot. ^cP < 0.001. PRNT: Prion protein testis specific; NC: Negative control; ZNF184: Zinc finger protein 184; HIPK2: Homeodomain interacting protein kinase 2.

Figure 8 prion protein testis specific acted as a sponge for zinc finger protein 184 to inhibit it and activated the expression of homeodomain interacting protein kinase 2. A: Anti-zinc finger protein 184 (ZNF184) RNA immunoprecipitation assays were performed after HCT116/L cells were transfected with prion protein testis specific (PRNT)-negative control (NC) or PRNT-mimics; B: Relative luciferase activities were evaluated after HCT116/L cells were transfected with homeodomain interacting protein kinase 2 (HIPK2)- wild-type or HIPK2- mutant and ZNF184 NC or ZNF184 mimics. ^cP < 0.001. PRNT: Prion protein testis specific; NC: Negative control; ZNF184: Zinc finger protein 184; HIPK2: Homeodomain interacting protein kinase 2; RIP: RNA immunoprecipitation.

Figure 9 Effect of prion protein testis specific knockdown or overexpression on the weight of xenografts derived from HCT116/L cells. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001.