Figure 3.

Validation of candidate miRNAs targeting NOTCH3. (A) Among four putative miRNAs, cotransfection of miR-150 reduced NOTCH3 luciferase activity in HEK293T cells. (B) miR-150 levels were significantly lower in lung adenocarcinoma tissues compared with adjacent normal tissues. miR-150 levels were negatively correlated with NOTCH3 mRNA expression (C), NCID3 protein (D), and HES1 protein (E) expression, both at lung adenocarcinoma tissues and normal tissues (n = 40). (F) qRT-PCR detection of miR-150 expression in HCC827GR6 cells transfected with miR-NC (group 1) or miR-150 mimic (group 2). (G) qRT-PCR and Western blot detection of NOTCH3 expression in HCC827GR6 cells transfected with miR-NC (group 1), miR-150 mimic (group 2), or cotransfected with miR-150 mimic and pEGFP-NOTCH3 (group 3). (H) The cell proliferation and (I) IC₅₀ of gefitinib were decreased in HCC827GR6 cells transfected with miR-150 mimic, but was reversed when cotransfected with pEGFP-NOTCH3.