Long non-coding RNA SOX21-AS1 modulates lung cancer progress upon microRNA miR-24-3p/PIM2 axis

Figures & data

Figure 1. High expression levels of SOX21-AS1 in lung cancer

(a) SOX21-AS1 levels in lung cancer and adjacent tissues were determined by RT-qPCR. ***P < 0.001 vs adjacent tissue. (b) The expression of SOX21-AS1 in normal lung epithelial cells (BEAS-2B) and lung cancer cells (H125, A549, NCI-H23, HCC827, and NCI-H1299) was also analyzed by RT-qPCR. **P < 0.01 vs BEAS-2B cells.

Figure 2. SOX21-AS1 targeted miR-24-3p in lung cancer

(a) The binding site between SOX21-AS1 and miR-24-3p as predicted by DIANA. (b) miR-24-3p levels in lung cancer and adjacent tissues were determined by RT-qPCR. **P < 0.01 vs adjacent tissue. (c) miR-24-3p expression levels in normal BEAS-2B cells and lung cancer cells were determined by RT-qPCR. **P < 0.01 vs BEAS-2B cells. (d) Luciferase reporter assays were used to assess the binding affinity between SOX21-AS1 and miR-24-3p. **P < 0.01 vs miR-24-3p mimic NC.

Figure 3. miR-24-3p inhibition reversed the decrease in cell proliferation induced by SOX21-AS1 downregulation

(a) The knockdown efficiency of siRNAs (siRNA-1 and siRNA-2) targeting SOX21-AS1 was examined by RT-qPCR. **P < 0.01 vs control. (b) The expression levels and inference efficacy of miR-24-3p mimics and an miR-24-3p inhibitor were measured by RT-qPCR. ***P < 0.001 vs control. (c-d) An MTT assay was used to evaluate the viability of A549 and HCC827 cells in four different groups. (e-f) Colony formation assays were used to assess the cell proliferation ability of A549 and HCC827 cells.

Figure 4. miR-24-3p inhibition decreased apoptosis induced by SOX21-AS1 downregulation

Apoptosis of A549 (a) and HCC827 (b) cells was analyzed by flow cytometry.

Figure 5. miR-24-3p depletion rescued the decrease in cell migration and invasion induced by SOX21-AS1 downregulation

(a-b) The ability of A549 and HCC827 cells to migrate and invade were assessed via transwell experiments.

Figure 6. miR-24-3p depletion reversed the decrease in tumor formation induced by SOX21-AS1 downregulation

(a) Tumor volumes of mice in the control, siRNA, and siRNA+miR-24-3p inhibitor groups. (b) Xenografted tumors from mice in the above three groups are shown.

Figure 7. miR-24-3p targeted PIM2, which was overexpressed in lung cancer

(a) The binding sequences of miR-24-3p and PIM2 were predicted using Targetscan. (b) RT-qPCR was used to analyze PIM2 expression in cancer and non-cancer tissues. ***P < 0.001 vs adjacent tissues. (c) Luciferase reporter assays were used to confirm the targeting of PIM2 by miR-24-3p. **P < 0.01 vs miR-24-3p mimic NC. (d) RT-qPCR analysis of PIM2 expression in cancer and normal cells. **P < 0.01 vs BEAS-2B cells.

Figure 8. miR-24-3p over-expression down-regulated the mRNA and protein expression levels of PIM2, inhibited proliferation, and promoted apoptosis

(a) RT-qPCR and western blotting were used to measure mRNA and protein expression levels in A549 cells. ***P < 0.001 vs control. (b) MTT experiments were used to analyze the proliferation of the two cell types. (c) Flow cytometry was used to detect the apoptotic ability of the two tumor cell types.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.