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. 2014 Oct 31;5(22):11669–11680. doi: 10.18632/oncotarget.2581

Figure 4. miR-9 down-regulates E-cadherin in ESCC cells.

Figure 4

(A) qRT-PCR and Western blotting results showed that miR-9 could effectively down-regulate E-cadherin expression in HK-miR-9 and 410-miR-9 cells compared with control cells. qRT-PCR results were expressed as mean ± S.E.M. of three independent experiments. **P < 0.001. GAPDH was set as an internal control in Western blot analysis. (B) Expressions of E-cadherin in both mRNA and protein levels were increased when miR-9 was silenced in KYSE30 and KYSE510 cells by a siRNA against miR-9, compared with scramble siRNA-treated cells. C, control scramble siRNA; si, siRNA against miR-9. **P < 0.01. (C) Luciferase assay was performed to confirm that miR-9 could target E-cadherin. miR-9 overexpressed cells and control cells were co-transfected with pMIR-REPORT-CDH1(3′-UTR)/empty vector with pRL-TK, and relative luciferase activity was detected. *P < 0.05. (D) Representative pictures of E-cadherin staining (by IHC) and miR-9 (by MISH) in a pair of ESCC tumor tissue and corresponding non-tumor tissue. Results showed that E-cadherin was down-regulated in miR-9 overexpressed tumor tissue. Original magnification: 20 × objective.