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. 2019 Nov 15;12:9795–9807. doi: 10.2147/OTT.S219440

Figure 3.

Figure 3

MTDH is a direct target gene of miR-618 in OS cells. (A) The 3′-UTR of the MTDH mRNA contains a potential miR-618–binding site. The mutant 3′-UTR region of the MTDH mRNA is also shown. (B) Plasmid pMIR-MTDH-3ʹ-UTR-wt or pMIR-MTDH-3ʹ-UTR-mut along with the miR-618 mimics or miR-NC was transfected into U2OS and HOS cells. After 48 h culture, the luciferase reporter assay was conducted to determine the luciferase activity. *P < 0.05 vs group miR-NC. (C, D) U2OS and HOS cells were transfected with the miR-618 mimics or miR-NC. The mRNA and protein levels of MTDH were measured by RT-qPCR and Western blot analysis, respectively. *P < 0.05 vs the miR-NC group. (E) Western blot analysis was conducted to detect MTDH protein expression in the human hFOB1.19 normal osteoblast cell line and four OS cell lines. (F) The mRNA expression of MTDH was detected in 41 pairs of OS tissue samples and ANT samples using RT-qPCR. *P < 0.05 vs ANTs. (G) Spearman correlation analysis was applied to assess the correlation between expression levels of miR-618 and MTDH mRNA in OS tissues. R2 = 0.3195, P = 0.0001.