The effect of mitotic death prostate cancer extracellular vesicles on PC3 cells
10.3760/cma.j.cn112330-20240203-00065
- VernacularTitle:裂亡前列腺癌细胞外囊泡对PC3细胞活性的影响
- Author:
Jian SHI
1
;
Yiming ZOU
;
Yangyang SUN
;
Xiaotong WU
;
Yuwei SHEN
;
Min FAN
Author Information
1. 苏州大学附属常州市第一人民医院泌尿外科，常州 213000
- Publication Type:Journal Article
- Keywords:
PC3 cell line;
Mitotic cell death;
Extracellular vesicles;
Therapy combined with adriamycin
- From:
Chinese Journal of Urology
2024;45(10):776-782
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of extracellular vesicles derived from mitotic cell death PC3 cells(MEV) on prostate cancer cells.Methods:After the phenotype of mitotic death was continuously induced by 1 μmol/L cisplatin for 5 days, nuclear damage was detected by immunofluorescence, cell stiffness was measured by Atomic Force Microscope, cell senescence marker(SA-β-Gal), proliferation, cycling, mitochondrial membrane potential, mitochondrial number by flow cytometry, and expression of senescence-secreting phenotype-associated factors including CDKNIA, CDKN2A, IL-6, IL-8, TNF-α, IFN-α/β/γ by qRT-PCR. Mitotic-death PC3 cells derived extracellular vesicles (MEV) were extracted, and the morphology and size of MEV were detected by electron microscopy and nanoparticle tracking analysis, and their stiffness and substance (β-actin and mtDNA) were separately detected by atomic force microscope and qRT-PCR. The effects of MEV on PC3 cells were further detected by immunofluorescence to detect phagocytosis, flow cytometry to detect proliferation and apoptosis, and qRT-PCR to detect the level of IFNα/β/γ. Finally, 50μg MEV were treated to PC3 cells combined with 15μmol/L adriamycin (Dox), and apoptosis was detected by flow assay.Results:Mitotic cell death PC3 cells and MEV were both obtained successfully.In comparison with normal PC3's, the number of MEV group was significantly increased[(4 530.9±353.6)×10 6(particle)/1×10 6 cell and (33.7±5.4)×10 6 particle/1×10 6 cell, P<0.01]. Additionally, the particle size of the extracellular vesicles were significantly smaller[(122.0±2.6)nm and (163.6±2.6)nm, P<0.01], along with significantly increased content of nuclear DNA[(111.0±20.7)/1×10 6 cell, P<0.01] and mtDNA[(26.2±3.8)/1×10 6 cell, P<0.01]. MEV were also easier to absorb by PC3 for their softness[(0.11±0.01)MPa and(0.18±0.01)MPa, P<0.01]. MEV could significantly induce PC3 cell apoptosis[(641.0±42.5)MFI and(351.7±37.0)MFI, P<0.01] and inhibit their proliferation[(1 523.0±64.9)MFI and(1 336.3±94.1)MFI, P<0.05].Besides, they did not affect the level of IFNβ but down-regulated IFNα/γmRNA level[(0.6±0.1)and(0.8±0.1), P<0.01].The combination of MEV and 15 μmol/L Dox can significantly promote PC3 cell apoptosis[(14 290.3±1315.9)MFI and(2 669.3±241.5)MFI, P<0.01]. Conclusions:Mitotic cell death PC3 cells can efficiently secrete DNA-rich flexible extracellular vesicles.The MEV were more easily taken up by PC3 and significantly inhibit its cellular activity and promote the anticancer effect of Dox.
