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Extracellular vesicles-encapsulated microRNA-10a-5p shed from cancer-associated fibroblast facilitates cervical squamous cell carcinoma cell angiogenesis and tumorigenicity via Hedgehog signaling pathway

Cancer Gene Therapy volume 28, pages 529–542 (2021)Cite this article

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Abstract

Cancer-associated fibroblast (CAF) secretes extracellular vesicle (EV)-encapsulated microRNAs (miRNAs) which have been underlined great promise for therapeutic target for diseases and cancers. Our study aimed to explore the role of EV-encapsulated miR-10a-5p from CAFs in angiogenesis in cervical cancer. Expression of miR-10a-5p in clinical samples of cervical cancer and cancer cells was detected by in situ hybridization and RT-qPCR. Results demonstrated that miR-10a-5p expression was upregulated in both cancer tissues and cells. CAFs and normal fibroblasts (NFs) from cervical cancer patient tissues were characterized under transmission electron microscopy, followed by EV isolation from CAFs. The EVs labeled with PKH67 were cultured with cervical squamous cell carcinoma (CSCC) cell line (SiHa) and HUVECs. Data indicated that CAF-EVs were internalized by cancer cells and promoted cell proliferation and tube formation. CAF-EVs then were transfected with miR-10a-5p inhibitor and then injected into nude mice. While injection of CAF-EVs promoted tumor growth and increased VEGFR and CD31 expression level, miR-10a-5p inhibitor-treated CAF-EVs resulted in decreased tumor volume and amount of vessel around tumor. Of note, dual-luciferase reporter gene assay and bioinformatic analysis indicated TBX5 as a target gene of miR-10a-5p. Moreover, EV-derived miR-10a-5p promoted angiogenesis in vivo and in vitro through activation of Hedgehog signaling via downregulation of TBX5. Taken altogether, miR-10a-5p in CAF-EVs promoted CSCC cell angiogenesis and tumorigenicity via activation of Hh signaling by inhibition of TBX5, providing insight into novel treatment based on miR-10a-5p against CSCC.

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Fig. 1: CAFs and EVs were successfully isolated.
Fig. 2: CAF-EVs facilitates tube formation of CSCC cells.
Fig. 3: High miR-10a-5p expression in CAF-EVs.
Fig. 4: CAF-EV-derived miR-10a-5p promotes angiogenesis.
Fig. 5: TBX5 is the direct target of miR-10a-5p.
Fig. 6: CAF-EV-miR-10a-5p promotes angiogenesis via targeting TBX5.
Fig. 7: CAF-EV-miR-10a-5p catalyzes angiogenesis through TBX5-mediated Hh signaling.
Fig. 8: Schematic diagram depicts that CAFs secreted EVs enriching miR-10a-5p and HUVECs internalized EVs. miR-10a-5p inhibited TBX5 expression and thus reduced inhibitory effect of TBX5 on Hh signaling.

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Funding

This study was supported by Sichuan Science and Technology Program of China (No. 2018SZ0248/2019YFS0404).

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Author notes

  1. The authors contributed equally: Xun Zhang, Yujue Wang

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Chengdu, 610072, China

    Xun Zhang, Yujue Wang, Bingyu Zou, Jie Mei & Zhao Wu

  2. Department of Obstetrics and Gynecology, Sichuan Provincial Hospital for Women and Children, Chengdu, 610045, China

    Xue Wang

  3. Department of Obstetrics and Gynecology, West China Second University Hospital, Chengdu, 610041, China

    Xue Peng

  4. Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Chengdu, 610041, China

    Xue Peng

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Contributions

Conceptualization: Xun Zhang and Yujue Wang; methodology: Xue Peng and Zhao Wu; software: Xue Wang; validation: Xue Peng; formal analysis: Xun Zhang; investigation: Bingyu Zou and Jie Mei; resources: Xun Zhang and Yujue Wang; data curation: Yujue Wang; writing—original draft: Xun Zhang and Yujue Wang; writing—review & editing: Xue Wang, Bingyu Zou, Jie Mei, Xue Peng, and Zhao Wu; visualization: Xue Peng; supervision: Zhao Wu; project administration: Zhao Wu.

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Correspondence to Xue Peng or Zhao Wu.

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The experiment was approved by the Ethics Committee of Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital and conducted in compliance with the Declaration of Helsinki. All individuals signed informed written consent documents. The experiments involving animals followed the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

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Zhang, X., Wang, Y., Wang, X. et al. Extracellular vesicles-encapsulated microRNA-10a-5p shed from cancer-associated fibroblast facilitates cervical squamous cell carcinoma cell angiogenesis and tumorigenicity via Hedgehog signaling pathway. Cancer Gene Ther 28, 529–542 (2021). https://doi.org/10.1038/s41417-020-00238-9

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