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. 2020 Jan 10;11(3):549–560. doi: 10.1111/1759-7714.13280

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© 2019 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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LncRNA NNT‐AS1 directly binds to miR‐22‐3p in NSCLC cells. (a) The putative binding sites of NNT‐AS1 and miR‐22‐3p were shown. (b and c) Dual‐luciferase reporter assay was conducted to detect the luciferase activity in H1299 and A549 cells cotransfected with WT‐NNT‐AS1 or MUT‐NNT‐AS1 and miR‐22‐3p mimic (miR‐22‐3p) or miR‐NC. (d and e) RNA pull‐down assay was performed to validate NNT‐AS1 targeted miR‐22‐3p. (f and g) The overexpression efficiency of NNT‐AS1 was measured in H1299 and A549 cells by qRT‐PCR analysis. (h and i) The expression of miR‐22‐3p was measured in H1299 and A549 cells transfected with sh‐NC, sh‐NNT‐AS1, pcDNA or NNT‐AS1. (j and k) The expression of miR‐22‐3p was detected in 37 paired NSCLC tissues and adjacent normal tissues using qRT‐PCR. (l) The expression of miR‐22‐3p was examined in BEAS‐2B cells and NSCLC cell lines (H1650, PC‐9, A549 and H1299) by qRT‐PCR. (m) The correlation between NNT‐AS1 and miR‐22‐3p in NSCLC tissues was analyzed. *P < 0.05. miR‐NC, miR‐22‐3p.

Inline graphic — LncRNA NNT‐AS1 directly binds to miR‐22‐3p in NSCLC cells. (a) The putative binding sites of NNT‐AS1 and miR‐22‐3p were shown. (b and c) Dual‐luciferase reporter assay was conducted to detect the luciferase activity in H1299 and A549 cells cotransfected with WT‐NNT‐AS1 or MUT‐NNT‐AS1 and miR‐22‐3p mimic (miR‐22‐3p) or miR‐NC. (d and e) RNA pull‐down assay was performed to validate NNT‐AS1 targeted miR‐22‐3p. (f and g) The overexpression efficiency of NNT‐AS1 was measured in H1299 and A549 cells by qRT‐PCR analysis. (h and i) The expression of miR‐22‐3p was measured in H1299 and A549 cells transfected with sh‐NC, sh‐NNT‐AS1, pcDNA or NNT‐AS1. (j and k) The expression of miR‐22‐3p was detected in 37 paired NSCLC tissues and adjacent normal tissues using qRT‐PCR. (l) The expression of miR‐22‐3p was examined in BEAS‐2B cells and NSCLC cell lines (H1650, PC‐9, A549 and H1299) by qRT‐PCR. (m) The correlation between NNT‐AS1 and miR‐22‐3p in NSCLC tissues was analyzed. *P < 0.05. miR‐NC, miR‐22‐3p.