Comprehensive Analysis of MicroRNA (miRNA) Targets in Breast Cancer Cells

Fan, Meiyun; Krutilina, Raisa I.; Sun, Jing; Sethuraman, Aarti; Yang, Chuan; Wu, Zhao-Hui; Yue, Junming; Pfeffer, Lawrence M.

doi:10.1074/jbc.m113.491803

Cited by 40 publications

(41 citation statements)

References 66 publications

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“…miRNAs have been increasingly recognized as key regulators of metastasis [27]. We previously showed that DROSHA knockdown promoted lung metastasis of basal-like breast cancer cells (MDA-MB-231) in an orthotopic xenograft model, suggesting a role of DROSHA-dependent miRNAs in repressing metastasis [28]. Therefore, we examined whether specific miRNAs play a role in regulating the expression of pro-metastatic genes.…”

Section: Resultsmentioning

confidence: 99%

“…To examine metastases in lungs, the dorsal surface of the left lung lobe was imaged using a fluorescent microscope at 10× magnification (NIKON, CFI60), and metastatic foci of mCherry expressing cells were quantified using ImageJ program [53]. The presence of tumor cells in the left lobes was further confirmed by Hematoxylin and Eosin (H&E) staining of formalin-fixed lung sections (10 μM thick) as described previously [28]. All animal studies adhered to protocols approved by the Institutional Animal Care and Use Committee of University of Tennessee Health Science Center.…”

Section: Methodsmentioning

confidence: 99%

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Systematic analysis of metastasis-associated genes identifies miR-17-5p as a metastatic suppressor of basal-like breast cancer

Fan

Sethuraman

Brown

et al. 2014

Breast Cancer Res Treat

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The purpose of this study is to identify metastasis- associated genes/signaling pathways in basal-like breast tumors. Kaplan–Meier analysis of two public meta-datasets and functional classification was used to identify genes/signaling pathways significantly associated with distant metastasis free survival. Integrated analysis of expression correlation and interaction between mRNAs and miRNAs was used to identify miRNAs that potentially regulate the expression of metastasis-associated genes. The novel metastatic suppressive role of miR-17-5p was examined by in vitro and in vivo experiments. Over 4,000 genes previously linked to breast tumor progression were examined, leading to identification of 61 and 69 genes significantly associated with shorter and longer DMFS intervals of patients with basal-like tumors, respectively. Functional annotation linked most of the pro-metastatic genes to epithelial mesenchymal transition (EMT) process and three intertwining EMT-driving pathways (hypoxia, TGFB and Wnt), whereas most of the anti-metastatic genes to interferon signaling pathway. Members of three miRNA families (i.e., miR-17, miR-200 and miR-96) were identified as potential regulators of the pro-metastatic genes. The novel anti-metastatic function of miR-17-5p was confirmed by in vitro and in vivo experiments. We demonstrated that miR-17-5p inhibition in breast cancer cells enhanced expression of multiple pro-metastatic genes, rendered cells metastatic properties, and accelerated lung metastasis from orthotopic xenografts. In contrast, intratumoral administration of miR-17-5p mimic significantly reduced lung metastasis. These results provide evidence supporting that EMT activation and IFN pathway inactivation are markers of metastatic progression of basal-like tumors, and members of miR-17, miR-200, and miR-96 families play a role in suppressing EMT and metastasis. The metastasis-associated genes identified in this study have potential prognostic values and functional implications, thus, can be exploited as therapeutic targets to prevent metastasis of basal-like breast tumors.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Systematic analysis of metastasis-associated genes identifies miR-17-5p as a metastatic suppressor of basal-like breast cancer

Fan

Sethuraman

Brown

et al. 2014

Breast Cancer Res Treat

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“…Some studies estimate miRNAs regulate ≥60% of the human protein-coding transcriptome(11). Importantly, single miRNAs can regulate entire cell signaling networks in a cell-context dependent manner(12). Since miRNAs can function as oncogenes or tumor suppressors, and can control the robustness of multiple cell signaling pathways(13,14), miRNAs that become dysregulated could promote a disease state, such as cancer.…”

Section: Introductionmentioning

confidence: 99%

miR-34a Silences c-SRC to Attenuate Tumor Growth in Triple-Negative Breast Cancer

et al. 2016

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Triple-negative breast cancer (TNBC) is an aggressive subtype with no clinically proven biologically targeted treatment options. The molecular heterogeneity of TNBC and lack of high frequency driver mutations other than TP53 have hindered the development of new and effective therapies that significantly improve patient outcomes. MicroRNAs (miRNAs), global regulators of survival and proliferation pathways important in tumor development and maintenance, are becoming promising therapeutic agents. We performed miRNA-profiling studies in different TNBC subtypes to identify miRNAs that significantly contribute to disease progression. We found that miR-34a was lost in TNBC, specifically within mesenchymal and mesenchymal-stem cell like subtypes, whereas expression of miR-34a targets were significantly enriched. Furthermore, restoration of miR-34a in cell lines representing these subtypes inhibited proliferation and invasion, activated senescence, and promoted sensitivity to dasatinib by targeting the proto-oncogene c-SRC. Notably, SRC-depletion in TNBC cell lines phenocopied the effects of miR-34a re-introduction, while SRC overexpression rescued the anti-tumorigenic properties mediated by miR-34a. miR-34a levels also increased when cells were treated with c-SRC inhibitors, suggesting a negative-feedback exists between miR-34a and c-SRC. Moreover, miR-34a administration significantly delayed tumor growth of subcutaneously and orthotopically implanted tumors in nude mice, and was accompanied by c-SRC downregulation. Finally, we found that miR-34a and SRC levels were inversely correlated in human tumor specimens. Together, our results demonstrate that miR-34a exerts potent anti-tumorigenic effects in vitro and in vivo, and suggests that miR-34a replacement therapy, which is currently being tested in human clinical trials, represents a promising therapeutic strategy for TNBC.

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“…For JNK knockdown, cells were transfected with ON-TARGETplus siRNA (20 nmol/L; Dharmacon), targeting JNK1 or JNK2 isoforms individually or in combination, and in order to activate JNK signaling, cells were transfected with constitutively active MKK7 and MEKK1 constructs. Immunoprecipitation (IP) of the AGO2 protein was carried out as previously described (36).…”

Section: Transient Transfections and Immunoprecipitationmentioning

confidence: 99%

Loss of miR-223 and JNK Signaling Contribute to Elevated Stathmin in Malignant Pleural Mesothelioma

Birnie

Yip

et al. 2015

Molecular Cancer Research

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Malignant pleural mesothelioma (MPM) is often fatal, and studies have revealed that aberrant miRNAs contribute to MPM development and aggressiveness. Here, a screen of miRNAs identified reduced levels of miR-223 in MPM patient specimens. Interestingly, miR-223 targets Stathmin (STMN1), a microtubule regulator that has been associated with MPM. However, whether miR-223 regulates STMN1 in MPM and the functions of miR-223 and STMN1 in this disease are yet to be determined. STMN1 is also regulated by c-Jun N-terminal kinase (JNK) signaling, but whether this occurs in MPM and whether miR-223 plays a role are unknown. The relationship between STMN1, miR-223, and JNK was assessed using MPM cell lines, cells from pleural effusions, and MPM tissue. Evidence indicates that miR-223 is decreased in all MPM tissue compared with normal/healthy tissue. Conversely, STMN1 expression was higher in MPM cell lines when compared with primary mesothelial cell controls. Following overexpression of miR-223 in MPM cell lines, STMN1 levels were reduced, cell motility was inhibited, and tubulin acetylation induced. Knockdown of STMN1 using siRNAs led to inhibition of MPM cell proliferation and motility. Finally, miR-223 levels increased while STMN1 was reduced following the re-expression of the JNK isoforms in JNK-null murine embryonic fibroblasts, and STMN1 was reduced in MPM cell lines following the activation of JNK signaling.Implications: miR-223 regulates STMN1 in MPM, and both are in turn regulated by the JNK signaling pathway. As such, miR-223 and STMN1 play an important role in regulating MPM cell motility and may be therapeutic targets.

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Comprehensive Analysis of MicroRNA (miRNA) Targets in Breast Cancer Cells

Cited by 40 publications

References 66 publications

Systematic analysis of metastasis-associated genes identifies miR-17-5p as a metastatic suppressor of basal-like breast cancer

Systematic analysis of metastasis-associated genes identifies miR-17-5p as a metastatic suppressor of basal-like breast cancer

miR-34a Silences c-SRC to Attenuate Tumor Growth in Triple-Negative Breast Cancer

Loss of miR-223 and JNK Signaling Contribute to Elevated Stathmin in Malignant Pleural Mesothelioma

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