https://orcid.org/0000-0003-2847-4040
Figures & data
Figure 1. LINC01094 expression is up-regulated in BC tissues
(a) With the GEPIA database, box plots were adopted for analyzing LINC01094 expression difference in BC and para-tumorous tissues. (b) Detection via qRT-PCR of LINC01094 expression in 54 cases of BC and para-cancerous tissues. (c) According to GEPIA database, the relationship between LINC01094 expression and the BC patient’s survival was analyzed. *P < 0.05, **P < 0.01 and ***P < 0.001.
Figure 2. LINC01094 has effects on cell proliferation, cell cycle progression and apoptosis
(a) Detection by qRT-PCR of LINC01094 expression in BC cell lines (MCF-7, L6, MDA-MB-231, MCF10CA1a and DU4475) and MCF-10A cells. (b) Detection via qRT-PCR of the transfection efficiency of LINC01094, si-LINC01094-1 and si-LINC01094-2 in MCF-7 and MDA-MB-231 cells. (c,d) BrdU and CCK-8 assays were performed to detect the effects of LINC01094 on MDA-MB-231 and MCF-7 cells’ proliferation. (e,f) The effects of LINC01094 on MDA-MB-231 and MCF-7 cell apoptosis and cell cycle were detected by flow cytometry assay. *P < 0.05, **P < 0.01 and ***P < 0.001
Figure 3. LINC01094 directly targets miR-340-5p in BC cells
(a) The subcellular location of LINC01094 was assessed via qRT-PCR after nucleocytoplasmic separation. (b) StarBase database predicted the binding site between LIN01094 and miR-340-5p. (c) MCF-7 and MDA-MB-231 cells were co-transfected with MUT LINC01094 or WT LINC01094 reporter with miR-340-5p inhibitors or mimics, respectively; the dual-luciferase reporter gene system was utilized to detect the luciferase activity. (d) RIP assay was employed to verify the binding relationship between LINC01094 and miR-340-5p. E. The effects of LINC01094 overexpression and knockdown on miR-340-5p expression in BC cells were detected via qRT-PCR. F. MiR-340-5p expression in 54 cases of BC and para-cancerous tissues was detected via qRT-PCR. G. Pearson correlation analysis of the correlation between LINC01094 and miR-340-5p expressions in BC tissues. *P < 0.05, **P < 0.01 and ***P < 0.001.
Figure 4. LINC01094 up-regulates E2F3 expression via sponging miR-340-5p
(a) StarBase database and TargetScan database were employed for predicting downstream miR-340-5p targets. (b, C) KEGG and GO databases were used to perform a pathway enrichment analysis of the target genes of miR-340-5p. (d) The online database StarBase was applied to predict the binding site of miR-340-5p to E2F3 mRNA 3ʹUTR. E. MUT E2F3 or WT E2F3 luciferase reporter vectors were co-transfected with miR-340-5p inhibitors or mimics into MCF-7 and MDA-MB-231 cells, and then the luciferase activity was detected. (f) Western blot was conducted to detect the regulatory effects of LINC01094 and miR-340-5p on E2F3 expression. (g) E2F3 mRNA expression in 54 pairs of BC tissues and para-cancerous tissues was examined through qRT-PCR. H-I. Pearson correlation analysis of the correlation among LINC01094, E2F3 mRNA and miR-340-5p expressions in BC tissues. *P < 0.05, **P < 0.01 and ***P < 0.001.
Figure 5. The impacts of LINC01094 and miR-340-5p of BC cell proliferation, cycle progression and apoptosis
A. LINC01094 overexpression plasmids, LINC01094 overexpression plasmids + miR-340-5p mimics and LINC01094 overexpressoin plasmids + miR-340-5p mimics + E2F3 overexpression plasmids were into MDA-MB-231 cells, respectively, and si-LINC01094-1, si-LINC01094-1 + miR-340-5p inhibitors and si-LINC01094-1 + miR-340-5p inhibitors + si-E2F3 were transfected into MCF-7 cells, respectively. The transfection efficiency was measured via qRT-PCR. B-C. BrdU and CCK-8 assays were utilized to detect the impacts of LINC01094, miR-340-5p, E2F3 on BC cell proliferation. D-E. Flow cytometry was utilized for detecting the impacts of LINC01094, miR-340-5p and E2F3 on BC cell apoptosis and cycle progression. *P < 0.05, **P < 0.01 and ***P < 0.001.
Figure 6. LINC01094 promoted the lung metastasis of MDA-MB-231 cells in vivo.
H&E staining was used to detect the metastatic nodule of the mice, which were injected with MDA-MB-231 cells transfected with LINC01094 overexpression plasmids or control plasmids, and the representative images were shown
Data availability statement
The data used to support the findings of this study are available from the corresponding author upon request.