Dan Zhang1, Dawei He1, Dian Li1, Bo Tang1, Dong Hu1, Wenhao Guo1, Zhang Wang1, Lianju Shen1, Guanghui Wei1. 1. ZHANG Dan, HE Dawei, LI Dian, TANG Bo, HU Dong, GUO Wenhao, WANG Zhang, SHEN Lianju, WEI Guanghui Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders; Chongqing Key Laboratory of Pediatrics Chongqing, 400014, China.
Abstract
OBJECTIVE: To investigate the effect of Piwil2-induced cancer stem-like cell (Piwil2-iCSC)-derived exosomes on the proliferation,migration and invasion of human umbilical cord blood-derived mesenchymal stem cells (hucMSCs). METHODS: Piwil2-iCSC-derived exosomes were isolated by ultracentrifugation and identified using transmission electron microscopy,nanoparticle tracking analysis and Western blotting.Exosome uptake assay was used to identify the pathway that Piwil2-iCSCderived exosomes utilized.HucMSCs were divided into control group,PBS intervention group and exosome intervention group,and CCK-8 assay,wound healing assay,Transwell assay,Western blotting and cell karyotype analysis were used to observe the proliferation,migration,invasion,expression levels of MMP2 and MMP9 proteins,and chromosome structure of hucMSCs. RESULTS: The diameter of Piwil2-iCSC-derived exosomes ranged from 50 nm to 100 nm,and most of them were oval or spherical capsules rich in CD9,CD63 and Piwil2 proteins.Exosomal uptake assay showed that the exosomes executed theirs functions after entering the cells.Compared with the control cells and PBS-treated cells,hucMSCs treated with the exosomes showed significantly increased number of proliferating cells (P<0.05) with accelerated healing rate (P<0.05 at 24 h;P<0.01 at 48 h),increased invasive cells (P<0.01),enhanced protein expressions of MMP2(P<0.05 vs PBS group;P<0.01 vs control group) and MMP9(P<0.05),but their karyotype still remained 46XY without any abnormalities. CONCLUSIONS: Piwil2-iCSC-derived exosomes can promote the proliferation,migration and invasion but does not cause cancer-like heterogeneity changes in hucMSCs.
OBJECTIVE: To investigate the effect of Piwil2-induced cancer stem-like cell (Piwil2-iCSC)-derived exosomes on the proliferation,migration and invasion of human umbilical cord blood-derived mesenchymal stem cells (hucMSCs). METHODS:Piwil2-iCSC-derived exosomes were isolated by ultracentrifugation and identified using transmission electron microscopy,nanoparticle tracking analysis and Western blotting.Exosome uptake assay was used to identify the pathway that Piwil2-iCSCderived exosomes utilized.HucMSCs were divided into control group,PBS intervention group and exosome intervention group,and CCK-8 assay,wound healing assay,Transwell assay,Western blotting and cell karyotype analysis were used to observe the proliferation,migration,invasion,expression levels of MMP2 and MMP9 proteins,and chromosome structure of hucMSCs. RESULTS: The diameter of Piwil2-iCSC-derived exosomes ranged from 50 nm to 100 nm,and most of them were oval or spherical capsules rich in CD9,CD63 and Piwil2 proteins.Exosomal uptake assay showed that the exosomes executed theirs functions after entering the cells.Compared with the control cells and PBS-treated cells,hucMSCs treated with the exosomes showed significantly increased number of proliferating cells (P<0.05) with accelerated healing rate (P<0.05 at 24 h;P<0.01 at 48 h),increased invasive cells (P<0.01),enhanced protein expressions of MMP2(P<0.05 vs PBS group;P<0.01 vs control group) and MMP9(P<0.05),but their karyotype still remained 46XY without any abnormalities. CONCLUSIONS:Piwil2-iCSC-derived exosomes can promote the proliferation,migration and invasion but does not cause cancer-like heterogeneity changes in hucMSCs.
Entities:
Keywords:
cancer stem cells; exosomes; human umbilical cord blood-derived mesenchymal stem cells; invasion; migration; proliferation
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