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. 2020 Mar 27:12:2301-2314.
doi: 10.2147/CMAR.S238051. eCollection 2020.

Acetyl-11-keto-β-boswellic Acid Inhibits Precancerous Breast Lesion MCF-10AT Cells via Regulation of LINC00707/miR-206 that Reduces Estrogen Receptor-α

Xuefeng Jiang #  1 , Yusheng Liu #  1 , Guijuan Zhang #  2 , Shujun Lin  1 , Naijun Yuan  1 , Jieyan Wu  1 , Xianxin Yan  1 , Yi Ma  3 , Min Ma  1   2
Affiliations

Acetyl-11-keto-β-boswellic Acid Inhibits Precancerous Breast Lesion MCF-10AT Cells via Regulation of LINC00707/miR-206 that Reduces Estrogen Receptor-α

Xuefeng Jiang et al. Cancer Manag Res. .

Abstract

Purpose: Acetyl-11-keto-β-boswellic acid (AKBA) has therapeutic effects on a range of diseases, including tumours. lncRNAs, as competing endogenous RNAs (ceRNAs), can interact with miRNAs to regulate the expression of target genes, which can affect the development of tumors. Here, we examined the effects of AKBA on breast precancerous lesions MCF-10AT cells.

Methods: The expression profiles of breast cancer (BC) tissue were collated from The Cancer Genome Atlas (TCGA), and the lncRNA-miRNA-mRNA ceRNA network was constructed. AKBA targets were predicted by network pharmacology. The expression of long intergenic nonprotein-coding RNA 707 (LINC00707), miR-206 and ER-α was determined by qRT-PCR. Cell viability, apoptosis and cycle were assessed by CCK-8 and flow cytometry. Protein levels were measured by Western blotting.

Results: A total of 3205 differentially expressed mRNAs, 104 miRNAs, and 605 lncRNAs were identified. The ceRNA network consisting of 9 lncRNAs, 15 miRNAs and 82 mRNAs was constructed. We found that LINC00707 was up-regulated and miR-206 was down-regulated in MCF-10AT cells. Transfected si-LINC00707 could inhibit cell proliferation, induce cell apoptosis and cycle arrest of MCF-10AT cells. In addition, network pharmacology predicted that AKBA may regulate the ESR1 in the treatment of BC. Our research demonstrated that AKBA could induce cell apoptosis and G1-phase arrest and inhibit ER-α expression via LINC00707/miR-206 in MCF-10AT cells.

Conclusion: AKBA inhibited MCF-10AT cells via regulation of LINC00707/miR-206 that reduces ER-α.

Keywords: ESR1; LINC00707; acetyl-11-keto-β-boswellic acid; breast precancerous lesion; ceRNA; miR-206.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
(A) volcano plot of differentially expressed genes (mRNAs, miRNAs and lncRNAs). Each point represents a gene (mRNA, miRNA or LncRNA). The red point in the plot represents up-regulated genes and green point represents down-regulated genes with statistical significance. (B) Heatmap of differentially expressed mRNAs, miRNAs and lncRNAs between BC and non-tumor tissues. Rows represent differently expressed genes and columns correspond to TCGA samples.
Figure 2
Figure 2
(A) The ceRNA network of lncRNAs-miRNAs-mRNAs in BC. (B) Biological Process analysis of the DEmRNAs in the ceRNA network. (C) PPI network of the DEmRNAs in the ceRNA network. (D) The ingredient-target-disease association network of AKBA.
Figure 3
Figure 3
Knockdown of LINC00707 inhibited the cells proliferation, induced apoptosis and cell cycle arrest of MCF-10AT cells. (A) The expression of LINC00707 and miR-206 in MCF-10A, MCF-10AT, MCF-7 cells. (B and C) The expression of ESR1 mRNA and ER-α protein in MCF-10A, MCF-10AT, MCF-7 cells. (D) Knockdown of LINC00707 inhibited the proliferation of MCF-10AT cells. (E) Inhibition of LINC00707 induced MCF-10AT cells apoptosis. (F) Inhibition of LINC00707 induced MCF-10AT cells cycle arrest. (G) The expression of Bcl2, Bax, Caspase-3, Cleaved Caspase-3, p27, Cyclin D1, CDK4 and CDK6 in MCF-10AT cells transfected with si-LINC00707. *P<0.05 compared with the normal blank control group.
Figure 4
Figure 4
LINC00707 functions as a ceRNA for miR-206 and indirectly modulates the expression of ER-α. (A) The binding sites between LINC00707 and miR-206. (B) The luciferase activity of MCF-10AT cells cotransfected with WT-LINC00707 or MUT-LINC00707 and miR-206 mimics. (C) The expression of miR-206 in MCF-10AT cells transfected with si-LINC00707. (D) The expression of ER-α protein in MCF-10AT cells transfected with si-LINC00707 or miR-206 mimics. *P<0.05 compared with the normal blank control group.
Figure 5
Figure 5
AKBA inhibited ER-α expression through LINC00707/miR-206 in MCF-10AT cells. (A) AKBA inhibited MCF-10AT cells growth. (B and C) Effect of AKBA on LINC00707, miR-206 and ESR1 in MCF-10AT cells. (D) Effect of AKBA on ER-α protein in MCF-10AT cells. *P<0.05 compared with the normal blank control group.
Figure 6
Figure 6
AKBA promoted apoptosis for MCF-10AT cells. (A) Flow cytometry assays of the apoptosis rates for MCF-10AT cells. (B) Effect of AKBA on the expression of Bcl2, Bax, Caspase-3 and Cleaved Caspase-3 in MCF-10AT cells. *P<0.05 compared with the normal blank control group.
Figure 7
Figure 7
AKBA induced G1-phase cell cycle arrest in MCF-10AT cells. (A) Flow cytometry assays of the cells cycle for MCF-10AT cells. (B) Effect of AKBA on the expression of p27, cyclin D1, CDK4 and CDK6 in MCF-10AT cells. *P<0.05 compared with the normal blank control group.
Figure 8
Figure 8
Schematic of the model. AKBA inhibits breast precancerous lesions MCF-10AT cells via regulation of LINC00707/miR-206/ER-α signalling.
See this image and copyright information in PMC

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