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. 2019 Aug 15;11:7739–7749. doi: 10.2147/CMAR.S207801

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© 2019 Huang et al.

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NNT-AS1 acted as a ceRNA by sponging miR-485 and regulated BCL9 expression indirectly.

Notes: (A) The predicted complementary binding sequences of NNT-AS1 3ʹ-UTR and miR-485. (B) The differential expression of miR-485 in CCA tissues and adjacent normal bile duct tissues was analyzed by qRT-PCR. (C) The miR-485 expression levels were investigated in CCA cells (CCLP1, QBC-939) and HIBEC by qRT-PCR. (D) Pearson’s correlation curve revealed the negative relevance between NNT-AS1 and miR-485 expression. (E) The expression levels of miR-485 in both CCLP1 and QBC-939 transfected with si-NNT-AS1 were evaluated by qRT-PCR. (F) Luciferase activity of 293T cells cotransfected with miR-485 mimic and luciferase reporters containing NNT-AS1-WT or NNT-AS1-MUT transcript were analyzed. (G) The predicted binding sites of miR-485 to the BCL9 sequence were shown. (H) Luciferase activity of 293T cells cotransfected with miR-485 mimic and luciferase reporters containing BCL9-WT or BCL9-MUT transcript were performed. (I) The BCL9 expression transfected with si-NNT-AS1 in CCLP1 and QBC-939 were analyzed by Western blot. The error bars in all graphs represented SD. *p<0.05, **p<0.01, ***p<0.001.

Abbreviations: NC, negative control; NNT-AS1, nicotinamide nucleotide transhydrogenase antisense RNA 1; MUT, mutation; WT, wild type.