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. 2021 Feb 3;14:751–761. doi: 10.2147/OTT.S240010

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© 2021 Gao et al.

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MCTP1-AS1 regulated SMAD7 expression via miR-650 in EC cells. (A) The binding scheme of miR-650 and SMAD7. (B) miR-650 interacts with SMAD7 by directly targeting verified by luciferase reporter assay. Data are mean ± SD of triplicate experiment (***p<0.001). (C) An inverse correlation is observed between mRNA expressions of miR-650 and SMAD7 in human EC tissues (n=60, p<0.0001). (D) qRT-PCR shows that mRNA expression level of SMAD7 is downregulated following miR-650 overexpression. Data are mean ± SD of triplicate experiment (**p<0.01). (E) Western blot shows that protein expression level of SMAD7 is downregulated following miR-650 overexpression. (F) A positive correlation is observed between mRNA expressions of MCTP1-AS1 and SMAD7 in human EC tissues (n=60, p<0.0001). (G) Si-SMAD7 is transfected into MCTP-AS1 overexpressed EC cells. CCK8 assay was used to determine the cell proliferation. Data are mean ± SD of triplicate experiment (****p<0.0001). (H) Wound healing assay was performed in MCTP1-AS1 overexpressed Ishikawa cell lines following transfection with si-SMAD7. (I) Transwell assay was used to measure the effect of si-SMAD7 transfection on cell migration and invasion in MCTP1-AS1 overexpressed Ishikawa cells. Data are mean ± SD of triplicate experiment (**p<0.01). (J) Western blot shows si-SMAD7 transfection significantly reverses the effect of MCTP1-AS1 on E-cadherin, N-cadherin and Vimentin expression in EC cells. (K) Western blot shows activation of TGF-β/SMAD pathway after transfecting si-SMAD7 in EC cells.