Study on the adhesion of E-cadherin between endothelial progenitor cells and bone marrow mesenchymal stem cells

doi:10.3877/cma.j.issn.2095-1221.2021.06.004

Abstract

Abstract:

Objective

To explore the adhesion of E-cadherin (E-cad) between endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) .

Methods

EPCs and MSCs were isolated and cultured from the bone marrow cavity of C57BL/6Jmice. Use Mito Tracker Red to label EPCs and DAPI to label MSCs. Immunofluorescence was used to examine the E-cad expression of the two cell lines. Cell adhesion assay was used to observe the adhesion of EPCs and MSCs in vitro. E-cad expression in EPCs and MSCs was silenced using E-cad siRNA, and siRNA transfection efficiency was determined by RT qPCR and Western blot. After successful transfection, the adhesion of EPCs and MSCs on the angiogenic Matrigel was observed. One way ANOVA was used for comparison between multiple groups, and LSD-t test was used for pairwise comparison between groups.

Results

(1) Compared with MSCs or EPCs, the expression of E-cad in the supernatant of the co-cultured of MSCs and EPCs in vitro (378.26±34.47, 564.72±41.58 vs 1087.28±101.92) was increased, the difference was statistically significant (P < 0.05) . (2) Compared with Control-siRNA, Mock and Negative, the expression of E-cad mRNA (0.97±0.07, 0.93±0.06, 1.00±0.03 vs 0.30±0.05) and E-cad protein (252.19 ±11.62, 223.70±13.66, 257.50±12.08 vs 31.74±4.08) of MSCs transfected with siRNA for 48 hours were decreased, the difference was statistically significant (P < 0.05) ; Compared with Control-siRNA, Mock and Negative, the expression of E-cad mRNA (0.93 ±0.07、0.88±0.08、1.00±0.02 vs 0.36±0.07) and E-cad protein (429.46±31.87、409.13±26.97、436.80±34.69 vs 54.03±15.05) of EPCs transfected with siRNA for 48 hours were decreased, the difference was statistically significant (P < 0.05) . (3) The adhesion of MSCs and EPCs were significantly inhibited after successful transfection of E-cad siRNA. (4) Adhesion and tube formation were significantly inhibited in the angiogenic Matrigel after E-cad of EPCs and MSCs was silenced.

Conclusions

E-cad could mediate the adhesion between EPCs and MSCs.

Key words: E-cadherin, Endothelial progenitor cells, Mesenchymal stem cells, Cell adhesion

Jie Xia, Zimeng Tang, Xiangwei Wu. Study on the adhesion of E-cadherin between endothelial progenitor cells and bone marrow mesenchymal stem cells[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2021, 11(06): 343-350.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2021.06.004

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Figures/Tables 6

References 29

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基因	引物序列（5'-3'）	预期扩增长度（bp）
E-cad	上游GCTGGACCGAGAGAGTTACC	160
	下游AGGCACTTGACCCTGATACG
β-actin（内参）	上游GTGCTATGTTGCTCTAGACTTCG	174
	下游ATGCCACAGGATTCCATACC

基因	引物序列（5'-3'）	预期扩增长度（bp）
E-cad	上游GCTGGACCGAGAGAGTTACC	160
	下游AGGCACTTGACCCTGATACG
β-actin（内参）	上游GTGCTATGTTGCTCTAGACTTCG	174
	下游ATGCCACAGGATTCCATACC

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