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Mesenchymal stem cell-derived exosomes block malignant behaviors of hepatocellular carcinoma stem cells through a lncRNA C5orf66-AS1/microRNA-127-3p/DUSP1/ERK axis

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Abstract

Mesenchymal stem cell (MSCs)-derived exosomes have been frequently used as useful tools in disease control. This research aimed to study the function of MSC-derived exosomes (Exo) in the stemness of cancer stem cells (CSCs) of hepatocellular carcinoma (HCC) and the molecular mechanism. Exo from the procured human bone marrow-MSCs were extracted and identified. CSCs from HCC cell lines were collected. The CSCs were treated with Exo, and then the proliferation, migration, invasion, angiogenesis-stimulating and self-renewal abilities of the Hep3B-CSCs and HuH7-CSCs were significantly reduced. C5orf66-AS1 was found as the most upregulated long noncoding RNAs (lncRNAs) in CSCs after Exo treatment. The integrated bioinformatic analyses and luciferase assays suggested that C5orf66-AS1 upregulated DUSP1 expression through sequestering microRNA-127-3p (miR-127-3p). Either artificial overexpression of miR-127-3p or silencing of DUSP1 blocked the inhibitory functions of Exo in the CSCs. DUSP1 inhibition increased the phosphorylation of ERK. Similar results were reproduced in vivo where Exo reduced the growth of xenograft formed by CSCs in nude mice, and this reduction was blocked upon miR-127-3p overexpression or DUSP1 silencing. To conclude, this research reported that MSC-derived Exo block malignant behaviors of HCC-sourced CSCs through a C5orf66-AS1/miR-127-3p/DUSP1/ERK axis.

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Data availability

The analyzed data sets generated during the study are available from the corresponding author on reasonable request.

Abbreviations

ANOVA:

Analysis of variance

ATCC:

American type culture collection

BCA:

Bicinchoninic acid

BCLC:

Barcelona clinic liver cancer

BM-MSCs:

Bone marrow-derived mesenchymal stem cells

CCK-8:

Cell counting kit-8

CSCs:

Cancer stem cells

CM:

Conditioned media

DMEM:

Dulbecco’s modified Eagle’s medium

DUSP1:

Dual-specificity phosphatase 1

Exo:

Exosomes

FBS:

Fetal bovine serum

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GEPIA:

Gene expression profiling interactive analysis

HCC:

Hepatocellular carcinoma

HUVECs:

Human umbilical vein endothelial cells

IgG:

Immunoglobulin G

IHC:

Immunohistochemical staining

LDA:

Limited-dilution assay

lncRNA:

Long noncoding RNA

mean ± SD:

Mean ± standard deviation

MT:

Mutant type

MTT:

3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

NC:

Negative control

NIH:

National Institutes of Health

NTA:

Nanoparticle tracking analysis

OD:

Optical density

PBS:

Phosphate-buffered saline

PFA:

Paraformaldehyde

RIPA:

Radio-immunoprecipitation assay

RT-qPCR:

Reverse transcription quantitative polymerase chain reaction

TEM:

Transmission electron microscope

WT:

Wild type

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Funding

This work was supported by Natural Science Foundation of Xinjiang Uygur Autonomous Region (No. 2017D01C306).

Author information

Author notes

  1. Hao Gu and Chao Yan contributed equally to this work.

Authors and Affiliations

  1. Department of Liver·Laparoscopic Surgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, Xinjiang, People’s Republic of China

    Hao Gu & Junjie Liu

  2. Department of Radiation Oncology, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University, Qingdao, 266035, Shandong, People’s Republic of China

    Chao Yan

  3. Department of Gastroenterology and Hepatology, Jinling Hospital Affiliated to Nanjing University School of Medicine, No. 305, East Zhongshan Road, Nanjing, 210002, Jiangsu, People’s Republic of China

    Haijun Wan & Lin Wu

  4. Department of Anorectal Surgery, Yantai Affiliated Hospital, Binzhou Medical College, Yantai, 264117, Shandong, People’s Republic of China

    Zhiqiang Zhu

  5. Department of Radiology Intervention, Jinling Hospital Affiliated to Nanjing University School of Medicine, No. 305, East Zhongshan Road, Nanjing, 210002, Jiangsu, People’s Republic of China

    Dazhi Gao

Authors
  1. Hao Gu
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Contributions

HG and CY: conceptualization, methodology, software, and writing—original draft preparation; LW: data curation; JJL: visualization and investigation; ZQZ: software and validation; DZG and HJW: conceptualization, writing—reviewing & editing, and writing—original draft preparation.

Corresponding author

Correspondence to Haijun Wan.

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The authors declare no competing interests.

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Signed informed consent was acquired from each participant. The research was approved by the Ethical Committee of the First Affiliated Hospital of Xinjiang Medical University (Approval number: K201807-05) and performed according to the Declaration of Helsinki.

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Gu, H., Yan, C., Wan, H. et al. Mesenchymal stem cell-derived exosomes block malignant behaviors of hepatocellular carcinoma stem cells through a lncRNA C5orf66-AS1/microRNA-127-3p/DUSP1/ERK axis. Human Cell 34, 1812–1829 (2021). https://doi.org/10.1007/s13577-021-00599-9

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