Figure 5.

Effect of ANRIL on cell proliferation.

Notes: (A) qRT-PCR analysis of ANRIL expression following treatment of H1975 cells with specific siRNA targeting ANRIL. (B) H1975 cells were transfected with si-ANRIL or si-NC. CCK8 assays were performed to determine the proliferation of H1975 cells. Data were expressed as the percentage of viable cells as follows: relative viability (%) = (A450_treated − A450_blank)/(A450_control − A450_blank) ×100%. (C) The ANRIL silencing inhibited DNA replication in H1975 cells compared to controls as determined by the EdU incorporation assay, Original magnifications: ×200. EdU-positive cell counts were analyzed by using software of the light microscope. (D) Changes in the proliferation marker, PCNA, were shown by Western blotting analysis and normalized to β-actin after si-ANRIL transfection. Relative protein expression was identified (n=3). (E) Knockdown of ANRIL could partly reverse c-Myc-induced growth promotion as determined with CCK-8 assays. Data represent the mean ± SD from three independent experiments. **P<0.01.

Abbreviations: CCK-8, Cell Counting Kit 8; EdU, ethynyl deoxyuridine; NC, negative control; OD, optical density; PCNA, proliferating cell nuclear antigen; qRT-PCR, quantitative real-time polymerase chain reaction; SD, standard deviation; si, small interfering.