Silibinin suppresses EMT-driven erlotinib resistance by reversing the high miR-21/low miR-200c signature in vivo

Cufí, Sílvia; Bonavia, Rosa; Vázquez-Martín, Alejandro; Oliveras‐Ferraros, Cristina; Corominas-Faja, Bruna; Cuyàs, Elisabet; Martín-Castillo, Begoña; Barrajón‐Catalán, Enrique; Visa, Joana; Segura‐Carretero, Antonio; Joven, Jorge; Bosch-Barrera, Joaquim; Micol, Vicente; Menendez, Javier A.

doi:10.1038/srep02459

Cited by 63 publications

(58 citation statements)

References 69 publications

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“…Using this model, which was developed in our laboratory, 46 we found that in the absence of second-site EGFR mutations, alternative activation of MET, AXL, or HER2, gain of secondary mutations in the KRAS, NRAS, or BRAF genes, or loss of the mutant delE746-A750 EGFR gene itself, the sole mechanism that accounted for the acquired resistance to erlotinib was a significant enrichment in EMT feature. 46,47 Here, we report for the first time an erlotinib-resistance transcriptomic signature that strongly suggests that erlotinib resistance can be explained by the acquisition of enhanced stem cell-like properties in EGFR-mutant NSCLC cell populations. Our study also demonstrates that erlotinibrefractory CSC cellular states, defined by the presence of very high levels of aldehyde dehydrogenase (ALDH) activity (i.e., ALDH bright cells), are exquisitely sensitive to the natural polyphenolic flavonoid silibinin, the active ingredient in milk thistle extracts that also exhibits anti-lung cancer activity.…”

Section: Introductionmentioning

confidence: 92%

“…We explored whether the flavonolignan silibinin, the only available experimental therapeutic that can circumvent erlotinib resistance in vivo, 47,48 can sensitize erlotinib-refractory ALDH bright cells to erlotinib treatment. To evaluate the effects of silibinin on the proportion of ALDH bright cells compared with vehicle treatment, erlotinib-refractory PC-9/Erl-R cells were treated (48 h) with increasing concentrations (50 and 100 μg/mL) of a milk thistle extract formulation that was enriched (30% w/w) with a water-soluble form of silibinin in complex with the amino-sugar Figure 2.…”

Section: The Natural Agent Silibinin Eliminates Erlotinib-resistant Amentioning

confidence: 99%

“…Our study also demonstrates that erlotinibrefractory CSC cellular states, defined by the presence of very high levels of aldehyde dehydrogenase (ALDH) activity (i.e., ALDH bright cells), are exquisitely sensitive to the natural polyphenolic flavonoid silibinin, the active ingredient in milk thistle extracts that also exhibits anti-lung cancer activity. [47][48][49][50][51] …”

Section: Introductionmentioning

confidence: 99%

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Stem cell-like ALDH^brightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

et al. 2013

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The enrichment of cancer stem cell (CSC)-like cellular states has not previously been considered to be a causative mechanism in the generalized progression of EGFR-mutant non-small cell lung carcinomas (NSCLC) after an initial response to the EGFR tyrosine kinase inhibitor erlotinib. To explore this possibility, we utilized a pre-clinical model of acquired erlotinib resistance established by growing NSCLC cells containing a TKI-sensitizing EGFR exon 19 deletion (ΔE746-A750) in the continuous presence of high doses of erlotinib. Genome-wide analyses using Agilent 44K Whole Human Genome Arrays were evaluated via bioinformatics analyses through GSEA-based screening of the KEGG pathway database to identify the molecular circuitries that were over-represented in the transcriptomic signatures of erlotinib-refractory cells. The genomic spaces related to erlotinib resistance included a preponderance of cell cycle genes (E2F1, - 2, CDC2, -6) and DNA replication-related genes (MCM4, - 5, - 6, - 7), most of which are associated with early lung development and poor prognosis. In addition, metabolic genes such as ALDH1A3 (a candidate marker for lung cancer cells with CSC-like properties) were identified. Thus, we measured the proportion of erlotinib-resistant cells expressing very high levels of aldehyde dehydrogenase (ALDH) activity attributed to ALDH1/3 isoforms. Using flow cytometry and the ALDEFLUOR® reagent, we confirmed that erlotinib-refractory cell populations contained drastically higher percentages (> 4500%) of ALDH(bright) cells than the parental erlotinib-responsive cells. Notably, strong decreases in the percentages of ALDH(bright) cells were observed following incubation with silibinin, a bioactive flavonolignan that can circumvent erlotinib resistance in vivo. The number of lung cancer spheres was drastically suppressed by silibinin in a dose-dependent manner, thus confirming the ability of this agent to inhibit the self-renewal of erlotinib-refractory CSC-like cells. This report is the first to show that: (1) loss of responsiveness to erlotinib in EGFR-mutant NSCLC can be explained in terms of erlotinib-refractory ALDH(bright) cells, which have been shown to exhibit stem cell-like properties; and (2) erlotinib-refractory ALDH(bright) cells are sensitive to the natural agent silibinin. Our findings highlight the benefit of administration of silibinin in combination with EGFR TKIs to target CSCs and minimize the ability of tumor cells to escape cell death in EGFR-mutant NSCLC patients.

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Section: Introductionmentioning

confidence: 92%

Section: The Natural Agent Silibinin Eliminates Erlotinib-resistant Amentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Stem cell-like ALDH^brightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

et al. 2013

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“…Likewise, reversal of cetuximab resistance by miR-200c is accompanied by the loss of EMT features in highly invasive and aggressive NSCLC cells [75]. Similarly, reversal of erlotinib resistance in EGFR-mutant NSCLC xenografts treated with a potent natural agent, silibilin, is associated with the loss of EMT-related features including low miR-200c and E-cadherin and high ZEB1, Snail, and N-cadherin expression [85]. However, in a similar study with erlotinib-resistant lung adenocarcinoma cells, overexpressed miR-200c regulates expression of EMT network proteins resulting in a slight increase in erlotinib sensitivity.…”

Section: Mir-200c Emt and Targeted Therapy Resistancementioning

confidence: 99%

miR-200c: a versatile watchdog in cancer progression, EMT, and drug resistance

et al. 2016

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MicroRNAs (miRNAs) are 20-22-nucleotide small endogenous non-coding RNAs which regulate gene expression at post-transcriptional level. In the last two decades, identification of almost 2600 miRNAs in human and their potential to be modulated opened a new avenue to target almost all hallmarks of cancer. miRNAs have been classified as tumor suppressors or oncogenes depending on the phenotype they induce, the targets they modulate, and the tissue where they function. miR-200c, an illustrious tumor suppressor, is one of the highly studied miRNAs in terms of development, stemness, proliferation, epithelial-mesenchymal transition (EMT), therapy resistance, and metastasis. In this review, we first focus on the regulation of miR-200c expression and its role in regulating EMT in a ZEB1/E-cadherin axis-dependent and ZEB1/E-cadherin axis-independent manner. We then describe the role of miR-200c in therapy resistance in terms of multidrug resistance, chemoresistance, targeted therapy resistance, and radiotherapy resistance in various cancer types. We highlight the importance of miR-200c at the intersection of EMT and chemoresistance. Furthermore, we show how miR-200c coordinates several important signaling cascades such as TGF-β signaling, PI3K/Akt signaling, Notch signaling, VEGF signaling, and NF-κB signaling. Finally, we discuss miR-200c as a potential prognostic/diagnostic biomarker in several diseases, but mainly focusing on cancer and its potential application in future therapeutics.

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“…Matrine, an alkaloid isolated from Sophora flavescens, downregulates miR-21 to induce overexpression of PTEN and inactivate Akt, leading to cell cycle arrest and apoptosis in MCF-7 breast cancer cells [136] . More recently, a study by Cufı΄ et al, reported that a water-soluble formulation of the flavolignan silibinin, the bioactive constituent of silymarin isolated from the dried fruits of the milk thistle, Silybum marianum plant [137] , was able to reverse the epithelial-to-mesenchymal transition (EMT)-related high miR-21/low miR-200c microRNA signature and repressed the mesenchymal markers SNAIL, ZEB, and N-cadherin observed in erlotinib-refractory non-small cell lung cancer (NSCLC) tumors [138] . Isothiocyanates, such as phenethyl isothiocyanate (PEITC) has been proposed to be a useful chemopreventive agent that can inhibit carcinogenic process.…”

Section: Other Natural Agentsmentioning

confidence: 99%

Modulation of miRNAs by Natural Agents: Nature’s way of dealing with cancer

Masika

Zhao

Hescheler

et al. 2015

RNA Dis

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Accumulating lines of evidence have revealed that microRNAs (miRNAs) play critical roles in many biological processes, such as carcinogenesis, angiogenesis, programmed cell death, cell proliferation, invasion, migration, and differentiation. They act either as tumor suppressors or oncogenes, and alteration in their expression patterns has been linked to onset, progression and chemoresistance of various cancers. Moreover, miRNAs are also crucial for the regulation of cancer stem cells (CSCs) self-renewal and proliferation as well as control of Epithelial-to-Mesenchymal Transition (EMT) of cancer cells. Therefore, exploitation of miRNAs as targets for cancer prevention and therapy could be a promising approach. Several experimental and epidemiologic studies have shown that dietary intake of natural agents such as baicalin, ginsenoside, curcumin, resveratrol, genistein, epigallocatechin-3-gallate (EGCG), indole-3-carbinol, 3,3΄-diindolylmethane (DIM) including antioxidants among others is inversely associated with the risk for cancer, demonstrating the inhibitory effects of natural agents on carcinogenesis. Additionally, the anticancer agents from natural agents have been found to inhibit the development and progression of cancer through the regulation of various cancer processes such as apoptosis, cell cycle regulation, differentiation, inflammation, angiogenesis, and metastasis. Importantly, natural agents could significantly impact the expression of tumor-suppressor and oncogenic miRNAs, regulate cellular signaling networks, inhibit cancer cell growth and cancer stem cell self-renewal. This is important for the normalization of altered cellular signaling mechanisms in cancer cells. This postulates a much broader use of natural agents in the prevention and/or treatment of various cancers in combination with conventional chemotherapeutics. However, more in vitro mechanistic experiments, in vivo animal studies, and clinical trials are warranted to realize the true value of natural agents in the prevention and/or treatment of cancer. Herein, we provide an overview of natural agents' modulation of miRNA expression as well as highlight the significance of these observations as potential new strategies in cancer therapies. This review will help us to understand how miRNAs are regulated by natural agents and also help in the development of effective and secure natural agents for therapeutic purposes.

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Silibinin suppresses EMT-driven erlotinib resistance by reversing the high miR-21/low miR-200c signature in vivo

Cited by 63 publications

References 69 publications

Stem cell-like ALDH^brightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

Stem cell-like ALDH^brightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

miR-200c: a versatile watchdog in cancer progression, EMT, and drug resistance

Modulation of miRNAs by Natural Agents: Nature’s way of dealing with cancer

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Silibinin suppresses EMT-driven erlotinib resistance by reversing the high miR-21/low miR-200c signature in vivo

Cited by 63 publications

References 69 publications

Stem cell-like ALDHbrightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

Stem cell-like ALDHbrightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

miR-200c: a versatile watchdog in cancer progression, EMT, and drug resistance

Modulation of miRNAs by Natural Agents: Nature’s way of dealing with cancer

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Product

Resources

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Stem cell-like ALDH^brightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin

Stem cell-like ALDH^brightcellular states in EGFR-mutant non-small cell lung cancer: A novel mechanism of acquired resistance to erlotinib targetable with the natural polyphenol silibinin