miR-34a mediates the multidrug resistance of breast cancer by regulating the expression of downstream gene
10.13602/j.cnki.jcls.2019.08.13
- VernacularTitle:miR-34a调控下游基因 FUT8 表达介导乳腺癌多药耐药的实验验证
- Author:
Xin WANG
1
;
Xiaolu MA
2
Author Information
1. Department of Clinical Laboratory, Dalian Sixth People's Hospital
2. Department of Clinical Laboratory, the First Affiliated Hospital of Dalian Medical University
- Publication Type:Journal Article
- Keywords:
miR-34a;
FUT8;
breast cancer;
multidrug resistance
- From:
Chinese Journal of Clinical Laboratory Science
2019;37(8):617-621
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To identify the therapeutic target of multidrug resistance of breast cancer by investigating the regulatory mechanism of the abnormal expression of miR-34a and FUT8 on the multidrug resistance of breast cancer.
Methods:The expression levels of miR-34a and FUT8 in MCF-7 and MCF-7/ADR cells were detected by the real-time PCR and western blot. The transfection efficiency of miR-34a in breast cancer cells was determined by the real-time PCR. The targeting relationship between miR-34a and FUT8 was predicted by the bioinformatics method, and further verified by the dual-luciferase reporter assay. After the expression of miR-34a was specifically regulated, the changes of FUT8 mRNA and protein levels in transfected cells were detected by the real-time PCR, Western blot and immunofluorescence staining, respectively. After the expression of miR-34a was specifically regulated, the proliferation and multidrug resistance of MCF-7 and MCF-7/ADR cells were determined by the CCK8 and immunofluorescence assays.
Results:The expression levels of miR-34a in MCF-7 cells were significantly higher than that in MCF-7/ADR cells ( P =0.002 6). The expression levels of FUT8 gene in MCF-7/ADR cells were significantly higher than that in MCF-7 cells ( P =0.001 6). FUT8 was the target of miR-34a regulating the drug resistance of breast cancer ( P =0.001 9). The up-regulation of miR-34a in MCF-7/ADR cells could significantly inhibit the expression of FUT8 , and the multidrug resistance and proliferation of MCF-7/ADR cells. While, the down-regulation of miR-34a increased the expression of FUT8 , and enhanced the multidrug resistance and proliferation of MCF-7 cells.
Conclusion:MiR-34a mediates the multidrug resistance of breast cancer by regulating the expression of downstream gene FUT8 .
