Fig 1: The mechanism of lncRNA 604 was detected as ceRNA. (A) The localization of lncRNA 604 was detected by FISH assay in the cells. (B) The expression of lncRNA 604 compared with the nuclear localization U1 and the cytoplasmic localization marker 18S. (C) LncRNA 604 spatial signal regulation network was generated by Targetscan software. (D) MiRNA 564 was regulated by lncRNA 604 by qRT-PCR in these bound miRNAs. (E) The binding site of lncRNA 604 and miRNA 564. (F) MiRNA 564 bounded the wild type of lncRNA 604 by the dual luciferin reporter gene. (G) The putative target genes of miRNA 564 was predicted by MiRanda (http://www.microrna.org). (H) The binding site of miRNA 564 and AEG-1. (I) miRNA 564 mimics reduced the luciferase activity of wt 3’UTR of AEG-1, while had no effect on mut 3’TUR of AEG-1. (J, K) MiRNA 564 could negatively regulate AEG-1 by qRT-PCR and WB [(J) RNA; (K) protein]. All **P < 0.05.
Fig 2: MTDH, EMT, and fibrosis markers protein expression after MTDH overexpression. (A) Representative bands from Western blot analyses of the levels of the MTDH in SV-HUC-1 cells after being transfected with MTDH overexpression NC plasmid and MTDH overexpression plasmid. (B) Representative bands from Western blot analyses of the levels of the MTDH, E-cadherin, vimentin, fibronectin, collagen ?, a-SMA protein in SV-HUC-1 cells after being treated with ketamine, ketamine and MTDH overexpression NC plasmid, ketamine and MTDH overexpression plasmid (treated for 48 h). (C) Relative levels of the MTDH, E-cadherin, vimentin, fibronectin, collagen ?, a-SMA protein compared to ß-actin. **p < 0.01; ***p < 0.0001; ****p < 0.0001. N=3.
Fig 3: LncRNA 604 promoted CRC cell chemoresistance in vivo. (A) The subcutaneous tumors were significantly smaller in low-expressed lncRNA 604 group while the high-expressed lncRNA 604 group failed to benefit, compared with their respective control groups. (B–D) AEG-1, NF-?b, and ERCC1 in subcutaneous tumor were detected by IHC, the trends were the same as WB in vitro (B) AEG-1; (C) NF-?b; (D) ERCC1). (E) The IRS staining score of AEG-1, NF-?b, and ERCC in all groups was graded. All **P < 0.05. The symbol **** represents the statistical significance of the two groups: black and red pillars.
Fig 4: Effects of overexpressing or silencing MTDH on cell invasion and migration in prostate normal and cancer cell lines. (A) Cell scratch wound assay was carried out to detect the effects of the overexpression or silencing of MTDH on the migration of the normal PWR-1E cells or 3 prostate cancer (PCa) cell lines (PC-3, LNCaP and DU145 cells). (B) Transwell assay was performed to detect the effects of the overexpression or silencing of MTDH on invasion of PWR-1E normal cells or 3 PCa cell lines (PC-3, LNCaP and DU145 cells). (C) Relative invasion rates are shown as bar diagrams. (D) Relative migration rates are shown as bar diagrams. Data are shown as the means ± SD from 3 independent experiments [**P<0.01, compared with control; ^^P<0.01, compared with mock/negative control (NC) groups]. MTDH, metadherin.
Fig 5: Expression of MTDH in prostate cancer (PCa) tissues and adjacent normal tissues, and its association with patient prognosis. (A) The TCGA database was used to analyze the association between MTDH and overall survival. (B) A total of 46 PCa and adjacent normal tissues were detected to determine the association between MTDH and overall survival by Kaplan-Meier analysis. (C) A total of 46 PCa tissues were detected for their mRNA expression of MTDH by RT-qPCR. (D) Immunohistochemistry was used to obverse the protein level of MTDH in two randomly selected patients (magnification: Upper panels, ×100; lower panels, ×200). MTDH, metadherin.
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