Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Aug 21:10:1397.
doi: 10.3389/fonc.2020.01397. eCollection 2020.

Exploring Specific miRNA-mRNA Axes With Relationship to Taxanes-Resistance in Breast Cancer

Danni Chen  1 , Chang Bao  2   3   4 , Feng Zhao  1 , Haogang Yu  1 , Guansheng Zhong  5 , Liang Xu  6 , Senxiang Yan  1
Affiliations

Exploring Specific miRNA-mRNA Axes With Relationship to Taxanes-Resistance in Breast Cancer

Danni Chen et al. Front Oncol. .

Abstract

Breast cancer is the most prevalent type of malignancy in women worldwide. Taxanes (paclitaxel and docetaxel) are widely applied as first-line chemotherapeutic agents, while the therapeutic effect is seriously limited by the development of drug resistance. In the present study, we screened out several miRNAs dysregulated in taxanes-resistant breast cancer samples and confirmed that two miRNAs (miR-335-5p and let-7c-5p) played a major role in cell proliferation, apoptosis, and chemo-resistance. In addition, the weighted gene co-expression network analysis (WGCNA) for potential target genes of miR-335-5p and let-7c-5p identified three hub genes (CXCL9, CCR7, and SOCS1) with a positive relationship to taxanes-sensitivity. Further, target relationships between miR-335-5p and CXCL9, let-7c-5p and CCR7/SOCS1 were confirmed by dual-luciferase reporter assays. Importantly, the regulatory functions of CXCL9, CCR7, and SOCS1 on proliferation and chemoresistance were validated. In conclusion, our study shed light on clinical theragnostic relationships between miR-335-5p/CXCL9, let-7c-5p/CCR7/SOCS1 axes, and taxanes-resistance in breast cancer.

Keywords: WCGNA; breast cancer; let-7c-5p/CCR7/SOCS1 axis; miR-335-5p/CXCL9 axis; taxanes-resistance.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Identification of dysregulated miRNAs relevant to taxanes-resistance in breast cancer. (A) The workflow of identification of miRNAs relevant to taxanes-resistance in breast cancer. (B) The heatmap showed dysregulated miRNAs in tumor tissues from comparation of taxanes-resistant and taxanes-sensitive groups. (C) Three selected miRNAs (miR-335-5p, let-7c-5p, and miR-99a-5p) were confirmed upregulation in tumor samples of taxanes-resistant breast cancer patients. (D) miR-335-5p, let-7c-5p, and miR-99a-5p were upregulated in breast cancer cell lines compared with breast cell line. (E) Three selected miRNAs were upregulated in paclitaxel-resistant breast cancer cell lines. (F) High expression of selected miRNAs predicted poorer overall survival in patients with breast cancer in the TCGA database. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 2
Figure 2
Validation biological functions of selected miRNAs in paclitaxel-resistant/sensitive cells. (A) Edu incorporation assays, (B) colony formation assays, (C) cell cycle assays were performed both in MCF-7/T and MCF-7 cells. (D) Apoptosis rate was detected in miR-335-5p/let-7c-5p-overexpressing cells and miR-335-5p/let-7c-5p-decreasing cells with or without paclitaxel. (E) The IC50 to paclitaxel and docetaxel was detected by MTT assays. All experiments were performed at least three times independently. The data were presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Scale bar: 500 μM.
Figure 3
Figure 3
The WGCNA for candidate target genes of miR-335-5p and let-7c-5p. (A) Clustering dendrogram of target genes of miR-335-5p and let-7c-5p, with dissimilarity based on Topological Overlap Measure (TOM). Genes were divided into five assigned merged modules, including 350 genes in gray module represented not co-expressed. (B) Interactive relationship analysis of co-expression genes in different modules. The brightness of yellow in the middle suggested a high-scale independence among the modules. (C) Eigengene adjacency heatmap showed the calculated eigengene of each module yielded in the above clustering analysis. (D) Module-trait associations between eigengenes in the module and taxanes-sensitivity of based on TCGA database. The yellow module showed highest correlation. (E) A scatterplot of Gene Significance (GS) and module membership (MM) in yellow module. There is a significantly positive correlation between GS and MM in the module. (F) Weight coefficient was calculated between pairwise genes in the yellow module and the hub genes were presented as different sizes of circles according to weight coefficient.
Figure 4
Figure 4
Identification of potential biological roles of the hub genes in the yellow module. (A,B) KEGG pathway enrichment and GO ontology analysis of all genes in the yellow module. (C) Overall survival curves based on hub genes expression of breast cancer patients in TCGA database. (D) Sub-network of candidate target genes and the corresponding upstream miRNAs.
Figure 5
Figure 5
CXCL9, CCR7/SOCS1 were validated as the direct target genes of miR-335-5p and let-7c-5p, respectively. (A) The dual-luciferase assays indicated miR-335-5p could directly target CXCL9, and let-7c-5p could directly bind to CCR7 and SOCS1. (B) The results of qRT-PCR and Western Blot indicated CXCL9 and CCR7 could be inhibited by miR-335-5p, CCR7, and SOCS1 could be inhibited by let-7c-5p both in mRNA and protein levels. (C) These three target genes were downregulated in paclitaxel-resistant cell lines (MCF-7/T and Bads-200). (D) CXCL9, CCR7, and SOCS1 were downregulated tumor samples of taxanes-resistant group in clinic. **P < 0.01, ***P < 0.001.
Figure 6
Figure 6
Identification of biological functions of CXCL9, CCR7, and SOCS1. (A) Edu incorporation assays, (B) colony formation assays, (C) Apoptosis rate was detected in CXCL9/CCR7/SOCS1-overexpressing MCF-7/T cells. (D) Cell cycle assays were performed in CXCL9/CCR7/SOCS1-overexpressing MCF-7/T cells. (E) The IC50 to paclitaxel and docetaxel was detected by MTT assays. All experiments were performed at least three times independently. The data were presented as the mean ± SD. **P < 0.01, ***P < 0.001. Scale bar: 500 μM.
Figure 7
Figure 7
Schematic model of miR-335-5p/CXCL9, let-7c-5p/CCR7/SOCS1 axes. Schematic model of miR-335-5p and let-7c-5p may regulate proliferation, apoptosis, and taxanes-resistance in breast cancer, by mediating CXCL9, CCR7, and SOCS1.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. (2020) 70:7–30. 10.3322/caac.21590 - DOI - PubMed
    1. Denny L, de Sanjose S, Mutebi M, Anderson BO, Kim J, Jeronimo J, et al. . Interventions to close the divide for women with breast and cervical cancer between low-income and middle-income countries and high-income countries. Lancet. (2017) 389:861–70. 10.1016/S0140-6736(16)31795-0 - DOI - PubMed
    1. Kesson EM, Allardice GM, George WD, Burns HJ, Morrison DS. Effects of multidisciplinary team working on breast cancer survival: retrospective, comparative, interventional cohort study of 13 722 women. BMJ. (2012) 344:e2718. 10.1136/bmj.e2718 - DOI - PMC - PubMed
    1. Goetz MP, Gradishar WJ, Anderson BO, Abraham J, Aft R, Allison KH, et al. . NCCN guidelines insights: breast cancer, version 3.2018. J Natl Compr Canc Netw. (2019) 17:118–26. 10.6004/jnccn.2019.0009 - DOI - PubMed
    1. Turner NC, Neven P, Loibl S, Andre F. Advances in the treatment of advanced oestrogen-receptor-positive breast cancer. Lancet. (2017) 389:2403–14. 10.1016/S0140-6736(16)32419-9 - DOI - PubMed