West China Journal of Stomatology ›› 2021, Vol. 39 ›› Issue (3): 313-319.doi: 10.7518/hxkq.2021.03.011

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Biological effects of exosome derived from Cal27 on normal human gingival fibroblasts

Jia Mei’e1(), Li Zhiyong2, Xu Kai1, Wang Yiheng1, Yu Fei1, He Xiangyi1()   

  1. 1.Dept. of Prosthodontics, School of Stomatology, Lanzhou University, Lanzhou 730000, China
    2.Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, China
  • Received:2020-04-15 Revised:2021-02-19 Online:2021-06-01 Published:2021-05-26
  • Contact: He Xiangyi E-mail:jiame12@lzu.edu.cn;hexy@lzu.edu.cn
  • Supported by:
    The National Natural Science Foundation of China(31572522);Scientific Research Foundation of School of Stomatology, Lanzhou University(lzukqky-2019-t01)

Abstract: Objective

The proliferation, migration capacity, and expression of activation-related proteins of NHGFs+Cal27-exo were determined by coculturing Cal27 exosome (Cal27-exo) with normal human gingival fibroblasts (NHGFs) to explore the effects of Cal27-exo on the activation and biological behavior of NHGFs.

Methods

Cal27-exo was extracted using supercentrifugation, and exosomes were identified using Western blot, transmission electron microscopy (TEM), and particle size detection. Cal27-exo was cocultured with NHGFs to detect the uptake of Cal27-exo by NHGFs, and the proliferation and migration capacity of NHGFs+Cal27-exo were detected using CCK8 and wound healing tests, respectively. The expression levels of NHGF activation-related proteins, i.e., matrix metalloproteinase-9 (MMP-9), fibroblast-activating protein (FAP), alpha smooth muscle actin (αSMA), and transforming growth factor-β (TGF-β), were detected using real-time quantitative polymerase chain reaction (qRT-PCR).

Results

Cal27-exo was extracted u-sing supercentrifugation, and Western blot showed the positive expression levels of Alix and CD63. TEM showed that Cal27-exo had a circular double-layer vesicle. The particle size was between 30 and 150 nm. Cal27-exo labeled with PKH67 entered NHGFs after the coculture method. The wound healing test showed that the migration capacity of NHGFs+Cal27-exo was stronger after the scratch compared with that of NHGFs. CCK8 results showed that the proliferation activity of NHGFs+Cal27-exo was enhanced. qRT-PCR results showed that the MMP-9 levels of NHGFs+Cal27-exo were upregulated, whereas the TGF-β and αSMA mRNA levels of NHGFs+Cal27-exo were downregulated (P<0.05).

Conclusion

The proliferation and migration ability of NHGFs+Cal27-exo are enhanced, and the mRNA expression of related proteins is changed. Cal27-exo can activate NHGFs, which suggests that Cal27-exo has potential significance in tumor invasion and metastasis.

Key words: oral cancer, exosome, cell proliferation, cell migration, fibroblast

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