Research Paper Volume 9, Issue 2 pp 475—486

Long intergenic non-protein coding RNA 00858 functions as a competing endogenous RNA for miR-422a to facilitate the cell growth in non-small cell lung cancer

Figure 4. LINC00858's pro-proliferative activity is in part through negative regulation of miRNA-422a, and then activation of KLK4 in NSCLC cells. (A) The 3'-UTR of KLK4 harbors one miR-422a cognate site. (B) Relative miR-422a expression after transfection with miR-NC and miR-422a. (C) Relative luciferase activity of reporter plasmids carrying wild-type or mutant KLK4 3'-UTR in A549 and SPC-A-1 cells co-transfected with miR-NC or miR-422a. (D) Relative KLK4 protein expression after transfection with miR-NC and miR-422a. (E) Relative KLK4 mRNA expression after transfection with miR-NC and miR-422a. (F) Statistical analysis of trypan blue staining. (G) Protein expression of KLK4 in Vector, miR-422a, pcDNA3.1-CT-GFP-KLK4, miR-422a plus pcDNA3.1-CT-GFP-KLK4, pcDNA3.1-CT-GFP-LINC00858, sh-KLK4, or pcDNA3.1-CT-GFP-LINC00858 +sh-KLK4 treated A549 and SPC-A-1 cells. Assays were performed in triplicate. *P < 0.05. Means ± SEM are shown. Statistical analysis was conducted using student One-Way ANOVA test.