Mobile website
Archive > Volume 20 Issue 2 > 2013,20(2):182-186. DOI:10.3872/j.issn.1007-385X.2013.02.009 Prev Next
Basic Research
Silencing HuR expression increases sensitivity of multidrug-resistant human breast cancer MCF-7/Adr cells to doxorubicin
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To investigate the effect of interference of human antigen R (HuR) expression on sensitivity of human multidrug-resistant human breast cancer MCF-7/Adr cell line to Doxorubicn. Methods: The shRNA expression vector targeting HuR gene (pGenesil-siHuR) has been constructed and stably transfected into human breast cancer MCF-7/Adr cell line. The expression level of MDR1 mRNA in MCF-7/Adr cells was assayed by real-time PCR. The P-gp protein (encoded by the MDR1 gene) expression were determined by Western blotting. The survival rate and IC50 of MCF-7/Adr cells to doxorubicin after pGenesil-siHuR transfection were evaluated by MTT method. The apoptosis rate of MCF-7/Adr cells was detected by flow cytometry. Results: Compared with untransfected MCF-7/Adr cells, the MDR1 mRNA（\[0.184±0.029\] vs \[1.203±0.026\]，P<0.01）and P-gp protein expressions（\[0.314±0.011\] vs \[0.796±0007\], P<0.01）were significantly reduced in pGenesil-siHuR transfected MCF-7/Adr cells (P<0.01). The IC50 of MCF-7/Adr cells to doxorubicin decreased from (148.2±2.3) nmol/L to (42.9±0.4) nmol/L after pGenesil-siHuR transfection. Compared with untransfected MCF-7/Adr cells, the ratio of cell apoptosis was significantly increased in pGenesil-siHuR transfected MCF-7/Adr cells （\[34.6±1.1\]% vs \[1.1±0.2\]%, P<0.01）after the treatment with doxorubicin. Conclusion: RNA interference of HuR can inhibit the expression of MDR1 gene and increase the sensitivity of multidrug-resistant breast cancer cells to doxorubicin.
Keywords:
Project Supported:
Project supported by the National Natural Science Foundation of China (No. 30901788, No. 81272619) , and the Natural Science Foundation of Shandong Province (No. ZR2010HQ038, No. ZR2010HM059)
Clc Number:
