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MicroRNA-20a-5p suppresses tumor angiogenesis of non-small cell lung cancer through RRM2-mediated PI3K/Akt signaling pathway

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Abstract

The current therapeutic strategies for non-small cell lung cancer (NSCLC) are limited and unsatisfactory. MicroRNAs (miRNAs) participate in tumor angiogenesis in NSCLC. The aim of this study was to investigate the role of miR-20a-5p (miR-20a) in human NSCLC metastasis. In the current study, bioinformatics analysis and RT-PCR were performed to examine the expression level of miR-20a in tissues of NSCLC patients and NSCLC cell lines, respectively. Western blot was performed to test the protein levels. Cell proliferation, migration and angiogenesis capacity were tested by 5-ethynyl-29-deoxyuridine (EdU) assay, transwell assay and tube formation assay, respectively. Dual-luciferase reporter assay (DLR) was used to confirm the interaction between miR-20a and paired ribonucleotide reductase regulatory subunit M2 (RRM2). We found that the expression of RRM2 was upregulated, while the expression of miR-20a was downregulated in cancer tissues compared with adjacent tissues in NSCLC patients. We also detected the expression level of RRM2 and miR-20a in NSCLC cell lines, showing A549 cell line exhibited the lowest expression level of miR-20a and highest expression level of RRM2. Overexpressed miR-20a not only dramatically suppressed NSCLC cells proliferation, endothelial cells migration and tube formation in vitro, but also inhibited tumor growth and angiogenesis in vivo. It was demonstrated that miR-20a suppressed NSCLC growth by inhibiting RRM2-mediated PI3K/Akt signaling pathway. These findings indicate that the novel identified miR-20a could function as a tumor suppressor in NSCLC through modulating the RRM2-mediated PI3K/Akt axis, and it could be a valid molecular target for NSCLC treatment.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

NSCLC:

Non-small cell lung cancer

miR-20a:

MiR-20a-5p

RRM2:

Ribonucleotide reductase regulatory subunit M2

LC:

Lung cancer

3′-UTRs:

3′ Untranslated regions

GEO:

Gene expression omnibus

DEGs:

Differentially expressed genes

BEAS-2B:

Human normal bronchial epithelial cell

HUVEC:

Human umbilical vein endothelial cells

DLR:

Dual-luciferase reporter assay

FBS:

Fetal bovine serum

RT:

Room temperature

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Funding

The work was funded by Henan Medical Science and Technology Project Plan (Joint construction, No. 2018020549).

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Authors and Affiliations

  1. Respiratory and Critical Illness Ward 1, Henan Chest Hospital, No. 1, Weiwu Road, Zhengzhou, 450000, Henan, China

    Junlei Han, Jianping Hu, Fang Sun, Hongzhi Bian, Bingxiang Tang & Xiang Fang

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  1. Junlei Han
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  2. Jianping Hu
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  3. Fang Sun
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  4. Hongzhi Bian
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  5. Bingxiang Tang
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Contributions

JLH participated in the design of the study, conducted the experiments, and drafted the manuscript. JPH designed the study, revised the manuscript and was responsible for authenticity of data. FS, HB, BT and XF collected and analyzed the data. All authors read and approved the final manuscript.

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Correspondence to Jianping Hu.

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The authors declare that they have no conflict of interest.

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The study was approved by Ethics committee of Henan Chest Hospital.

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Han, J., Hu, J., Sun, F. et al. MicroRNA-20a-5p suppresses tumor angiogenesis of non-small cell lung cancer through RRM2-mediated PI3K/Akt signaling pathway. Mol Cell Biochem 476, 689–698 (2021). https://doi.org/10.1007/s11010-020-03936-y

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  • DOI: https://doi.org/10.1007/s11010-020-03936-y

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