Transfer of siRNA against CD73 suppresses human breast cancer cell line MB-MDA-231 adhesion to extracellular matrix
- VernacularTitle:siRNA干扰CD73表达抑制人乳腺癌细胞MB-MDA-231与细胞外基质的黏附
- Author:
Xiuling ZHI
;
Tingting ZHOU
;
Li WANG
;
Fengdi ZHAO
;
Lianhua YIN
;
Ping ZHOU
- Publication Type:Journal Article
- Keywords:
5'-nucleotidase;
Adenosine;
RNA interference;
Cell adhesion
- From:
Chinese Journal of Pathophysiology
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To study the effect of CD73 on breast cancer cell line MB-MDA-231 adhesion to extracellular matrix.METHODS: ① CD73 siRNA plasmid was constructed and transfected into MB-MDA-231 cells by lipofectamine 2000.② The transfection efficiency was analyzed by flow cytometry using eGFP as a marker gene.Stable transfected MB-MDA-231 cells were selected using G418.③ RT-PCR and Western blotting analysis of CD73 expression in MB-MDA-231 cells was performed.④ The effects of CD73 on MB-MDA-231 cells adhesion to extracellular matrix were assessed by cell adhesion assay.RESULTS: ① Transfection of MB-MDA-231 cells with CD73 siRNA plasmid significantly inhibited CD73 expression at mRNA level.The efficiency was up to 91%.② Transfection of MB-MDA-231 cells with CD73 siRNA plasmid significantly inhibited CD73 expression at a protein level.The efficiency was up to 79.3%.③ Treatment of MB-MDA-231 cells with CD73 siRNA resulted in diminished adhesion to extracellular matrix.CONCLUSION: This study demonstrates that CD73 siRNA effectively inhibits CD73 gene expression in MB-MDA-231 cells leading to adhesion to extracellular matrix suppression.
