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pmid:29531296

miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma.

Ailiang Zeng · Jianxing Yin · Yan Li · Rui Li · Zheng Wang · Xu Zhou · Xin Jin · Feng Shen · Wei Yan · Yongping You

Cell death & disease · 2018 doi:10.1038/s41419-018-0343-1

Paper

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Experiment

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Data

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Analysis

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title
miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma.
doi
10.1038/s41419-018-0343-1
pmid
29531296
authors array · 10 items
[0]
Ailiang Zeng
[1]
Jianxing Yin
[2]
Yan Li
[3]
Rui Li
[4]
Zheng Wang
[5]
Xu Zhou
[6]
Xin Jin
[7]
Feng Shen
[8]
Wei Yan
[9]
Yongping You
journal
Cell death & disease
year
2018
abstract
Therapeutic application of microRNAs (miRNAs) in Wnt-driven glioma has been valuable; however, their specific roles and mechanisms have not been completely investigated. Real-time quantitative PCR (RT-qPCR) was used to analyse the expression of microRNA-129-5p (miR-129-5p) in human glioma samples. Cell-Counting Kit 8 (CCK-8), flow cytometry, EdU, angiogenesis, Transwell invasion, wound healing, in vitro 3D migration and neurosphere formation assays were employed to assess the role of miR-129-5p in glioblastoma multiforme (GBM) cells. Moreover, we performed the luciferase reporter assay and the RNA-ChIP (chromatin immunoprecipitation) assay to confirm whether Wnt5a was a direct target of miR-129-5p. We also confirmed the correlation between the expression profile of miR-129-5p and Wnt5a in glioma patients from the Chinese Glioma Genome Atlas (CGGA) and investigated the overall survival of GBM patients using two data sets, namely, TCGA and GSE16011, according to their Wnt5a expression status. MiR-129-5p expression levels were downregulated and inversely correlated with Wnt5a expression levels in CGGA glioma patients. Restored expression of miR-129-5p blocked GBM cell proliferation, invasion, migration, angiogenesis, neurosphere formation and resistance to temozolomide. We reported that miR-129-5p directly targeted Wnt5a in glioma. Furthermore, we observed that overexpression of miR-129-5p inhibited the expression of Wnt5a, thus blocking the protein kinase C(PKC)/ERK/NF-κB and JNK pathways. Inhibiting Wnt5a rescued the effects of miR-129-5p loss and increased Wnt5a expression was associated with reduced overall survival of GBM patients. We also demonstrated the inhibitory effect of miR-129-5p on tumour growth in GBM using an in vivo model. The miR-129-5p/Wnt5a-axis-mediated PKC/ERK/NF-κB and JNK pathways have therapeutic potential in GBM treatment.
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To fully explore the association of Wnt5a expression with survival in all malignant gliomas, we conducted univariate and multivariate Cox regression analysis to assess the prognostic independence of Wnt5a expression among other factors. Increasing Wnt5a expression was correlated with a worse outcome in malignant glioma patients in all included data sets (Fig. 7l and Supplementary Table 1 and 2). Increasing Wnt5a was independently correlated with a worse outcome in malignant glioma patients in the GSE16011 data set (Fig. 7l).
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sections[9] "miR-129-5p inhibits tumour growth in vivo and elevated Wnt5a is associated with decreased survival" paragraph 5
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To fully explore the association of Wnt5a expression with survival in all malignant gliomas, we conducted univariate and multivariate Cox regression analysis to assess the prognostic independence of Wnt5a expression among other factors. Increasing Wnt5a expression was correlated with a worse outcome in malignant glioma patients in all included data sets (Fig. 7l and Supplementary Table 1 and 2). Increasing Wnt5a was independently correlated with a worse outcome in malignant glioma patients in the GSE16011 data set (Fig. 7l).
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We first analysed miR-129-5p levels in data from 491 GBM patients using the TCGA database. The Spearman test of correlation of miR-129-5p expression with Wnt5a transcript expression from both CGGA mRNA sequences and array data sets also revealed a statistically significant inverse correlation between miR-129-5p and Wnt5a expression levels (Fig. 2h, i).
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sections[3] "miR-129-5p expression is downregulated in glioma" paragraph 2
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We first analysed miR-129-5p levels in data from 491 GBM patients using the TCGA database.
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sections[4] "Wnt5a is a direct target of miR-129-5p" paragraph 2
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The Spearman test of correlation of miR-129-5p expression with Wnt5a transcript expression from both CGGA mRNA sequences and array data sets also revealed a statistically significant inverse correlation between miR-129-5p and Wnt5a expression levels (Fig.
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The paper used univariate and multivariate Cox regression analysis to assess the prognostic association of Wnt5a expression, with survival data obtained from the GSE16011 and other glioma datasets. The headline result was that miR-129-5p directly targets Wnt5a and suppresses malignant GBM phenotypes.
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To fully explore the association of Wnt5a expression with survival in all malignant gliomas, we conducted univariate and multivariate Cox regression analysis to assess the prognostic independence of Wnt5a expression among other factors.
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sections[9] "miR-129-5p inhibits tumour growth in vivo and elevated Wnt5a is associated with decreased survival" paragraph 5
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Increasing Wnt5a was independently correlated with a worse outcome in malignant glioma patients in the GSE16011 data set
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We reported that miR-129-5p directly targeted Wnt5a in glioma.
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The paper used univariate and multivariate Cox regression analysis to assess the prognostic association of Wnt5a expression, with survival data obtained from the GSE16011 and other glioma datasets. The headline result was that miR-129-5p directly targets Wnt5a and suppresses malignant GBM phenotypes.
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miR-129-5p on tumour growth in GBM using an in vivo model. The miR-129-5p/Wnt5a-axis-mediated PK
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To fully explore the association of Wnt5a expression with survival in all malignant gliomas, we conducted univariate and multivariate Cox regression analysis to assess the prognostic independence of Wnt5a expression among other factors. Increasing Wnt5a expression was correlated with a worse outcome in malignant glioma patients in all included data sets (Fig. 7l and Supplementary Table 1 and 2). Increasing Wnt5a was independently correlated with a worse outcome in malignant glioma patients in the GSE16011 data set (Fig. 7l).
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Complete file
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Real-time quantitative PCR (RT-qPCR) was used to analyse the expression of microRNA-129-5p (miR-129-5p) in human glioma samples. Cell-Counting Kit 8 (CCK-8), flow cytometry, EdU, angiogenesis, Transwell invasion, wound healing, in vitro 3D migration and neurosphere formation assays were employed to assess the role of miR-129-5p in glioblastoma multiforme (GBM) cells. Moreover, we performed the luciferase reporter assay and the RNA-ChIP (chromatin immunoprecipitation) assay to confirm whether Wnt5a was a direct target of miR-129-5p. We also confirmed the correlation between the expression profile of miR-129-5p and Wnt5a in glioma patients from the Chinese Glioma Genome Atlas (CGGA) and investigated the overall survival of GBM patients using two data sets, namely, TCGA and GSE16011, according to their Wnt5a expression status. MiR-129-5p expression levels were downregulated and inversely correlated with Wnt5a expression levels in CGGA glioma patients. Restored expression of miR-129-5p blocked GBM cell proliferation, invasion, migration, angiogenesis, neurosphere formation and resistance to temozolomide. We reported that miR-129-5p directly targeted Wnt5a in glioma. Furthermore, we observed that overexpression of miR-129-5p inhibited the expression of Wnt5a, thus blocking the protein kinase C(PKC)/ERK/NF-κB and JNK pathways. Inhibiting Wnt5a rescued the effects of miR-129-5p loss and increased Wnt5a expression was associated with reduced overall survival of GBM patients. We also demonstrated the inhibitory effect of miR-129-5p on tumour growth in GBM using an in vivo model. 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Cell-Counting Kit 8 (CCK-8), flow cytometry, EdU, angiogenesis, Transwe"], "order_basis": "paper_text_order:357", "id": "assay:2"}, {"role": "data_generation", "definition_id": "flow_cytometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["glioma samples. Cell-Counting Kit 8 (CCK-8), flow cytometry, EdU, angiogenesis, Transwell invasion, woun"], "order_basis": "paper_text_order:365", "id": "assay:3"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.77, "evidence": ["miR-129-5p on tumour growth in GBM using an in vivo model. The miR-129-5p/Wnt5a-axis-mediated PK"], "order_basis": "paper_text_order:1790", "id": "assay:4"}, {"role": "analysis", "definition_id": "survival_analysis", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["cox_ph"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["To fully explore the association of Wnt5a expression with survival in all malignant gliomas, we conducted univariate and multivariate Cox regression analysis to assess the prognostic independence of Wnt5a expression among other factors. Increasing Wnt5a expression was correlated with a worse outcome in malignant glioma patients in all included data sets (Fig. 7l and Supplementary Table 1 and 2). Increasing Wnt5a was independently correlated with a worse outcome in malignant glioma patients in the GSE16011 data set (Fig. 7l)."], "order_basis": "consumes_earlier_output", "id": "assay:5"}]}