Figures & data
Table 1 Correlation Between GATA6-AS1 Expression and Clinical Pathological Factors of Patients
Figure 1 LncRNA GATA6-AS1 was decreased in lung cancer. (A) The expression of GATA6-AS1 in multiple cancers and normal tissues were investigated in TCGA datasets and GTEx project using GEPIA software. The expression of GATA6-AS1 in tumors were presented as red dots, while green dots represented GATA6-AS1 expression in normal tissues. (B) GATA6-AS1 levels were detected in 100 pairs of lung cancer tumor tissues and matched normal tissues from patients. (C) Comparison of GATA6-AS1 levels in lung tumors of non-metastasis and those of metastasis. (D) Kaplan–Meier Plotter analysis was performed to study the association between GATA6-AS1 expression and OS of patients with lung cancer (n=1144). ***p<0.001.
Figure 2 Ectopic expression of GATA6-AS1 repressed lung cancer cell invasion and proliferation. (A) Detection of GATA6-AS1 in a panel of lung cancer cell lines (NCI-H1975, A549, NCI-H460, and NCI-H1299) and normal human bronchial epithelial cell line 16-HBE by RT-qPCR. (B) RT-qPCR was used to detect GATA6-AS1 level in NCI-H1975 and A549 cells transfected with pcDNA3 or pcDNA3-GATA6-AS1. (C) Transwell assay was used to study the cell number invaded through the membrane in NCI-H1975 and A549 cells transfected with pcDNA3 or pcDNA3-GATA6-AS1. D-E. CCK-8 assay was used to study the cell proliferation ability in NCI-H1975 (D) and A549 (E) cells transfected with pcDNA3 or pcDNA3-GATA6-AS1. *p<0.05; **p<0.01; ***p<0.001.
Figure 3 GATA6-AS1 downregulated miR-324-5p in lung cancer cells. (A) Analysis by miRDB software showed a binding site for miR-324-5p in GATA6-AS1. (B) Overexpression of GATA6-AS1 decreased miR-324-5p expression in NCI-H1975 and A549 cells. (C) miR-324-5p inhibitor decreased miR-324-5p level in NCI-H1975 and A549 cells as measured in RT-qPCR. (D) miR-324-5p inhibitor increased GATA6-AS1 level in NCI-H1975 and A549 cells as measured in RT-qPCR. ***p<0.001.
Figure 4 GATA6-AS1 sponged miR-324-5p. (A) Construction of luciferase plasmid containing GATA6-AS1-wild type or GATA6-AS1-Mutant. (B) Transfection of miR-324-5p mimic increased miR-324-5p level in NCI-H1975 and A549 cells. (C and D) miR-324-5p mimic decreased luciferase activity of GATA6-AS1-wild type but not in GATA6-AS1 in NCI-H1975 (C) and A549 (D) cells. **p<0.01; ***p<0.001.
Figure 5 GATA6-AS1 promoted FBXO11 expression. (A and B) Western blotting showed that GATA6-AS1 increased FBXO11 and decreased SNAIL protein expression in NCI-H1975 (A) and A549 (B) cells. (C) GATA6-AS1 increased FBXO11 mRNA expression in NCI-H1975 and A549 cells. (D and E) GATA6-AS1 increased E-cadherin and decreased N-cadherin mRNA expression in NCI-H1975 (D) and A549 (E) cells. **p<0.01; ***p<0.001.
Figure 6 GATA6-AS1 regulated FBXO11 via sponging miR-324-5p. (A and B) Western blotting showed that miR-324-5p mimic could reverse the upregulation of FBXO11 and downregulation of SNAIL protein expression in NCI-H1975 (A) and A549 (B) cells transfected with GATA6-AS1. (C) In NCI-H1975 and A549 cells, miR-324-5p could reverse the upregulation of FBXO11 mRNA expression by GATA6-AS1. (D) Sequences of FBXO11 3ʹUTR-wild type and 3ʹUTR-Mutant. E-F. In the luciferase reporter assay, GATA6-AS1 could attenuate the reduction of FBXO11 3ʹUTR-wild type luciferase activity in NCI-H1975 (E) and A549 (F) cells transfected with miR-324-5p mimic. ***vs pcDNA3 + miR-NC, p<0.001; ###vs GATA6-AS1 + miR-NC, p<0.001.
Figure 7 GATA6-AS1 regulated lung cancer cell invasion and proliferation via sponging miR-324-5p (A). Transwell assay showed that miR-324-5p mimic could reverse the inhibitory effect of GATA6-AS1 on cell invasion in NCI-H1975 and A549 cells. (B and C) CCK-8 assay showed that miR-324-5p mimic could reverse the inhibitory effect of GATA6-AS1 on cell proliferation in NCI-H1975 (B) and A549 (C) cells. **vs pcDNA3 + miR-NC, p<0.01; ***vs pcDNA3 + miR-NC, p<0.001; #vs GATA6-AS1 + miR-NC, p<0.05; ##vs GATA6-AS1 + miR-NC, p<0.01; ###vs GATA6-AS1 + miR-NC, p<0.001.
