Evaluation of Cisplatin and Cisplatin-loaded Magnetic Iron Oxide Nanoparticles on BCL2 and BAX genes in the Breast Cancer T47D Cell Line

Authors
M. J. Mokhtari 1
Z. Hoseineian 2
F. Koohpeima 3
A. Akbarzadeh 4
M. Hashemi 5
R. Mahdian 6
A. R. Kamyab 7
H. Mohammadi 8
1 Assistant Professor, Department of Biology, Zarghan Branch, Islamic Azad University, Zarghan, Iran
2 M.Sc., Department of Pilot Biotechnology, Pasteur Institute of Iran, Tehran, Iran
3 Assistant Professor, Department of Operative Dentistry, School of Dentistry, Shiraz University of Medical Science, Shiraz, Iran
4 Professor, Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran
5 Associated Professor, Department of Genetics, Tehran Medical Branch, Islamic Azad University, Tehran, Iran
6 Assistant Professor, Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
7 M. Sc, Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
8 M. Sc, Young Researchers Club, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran
Abstract
Objective: Breast cancer is the second leading cause of cancer death in women. Cisplatin is a traditional cancer drug commonly used in chemotherapy for killing cancer cells. Modulation at the mRNA levels of apoptotic related genes often correlate with the sensitivity of various types of cancer cells to chemotherapeutic agents. Nanoparticulate drug delivery systems are being developed to effectively deliver smaller doses of chemotherapeutic agents and control drug distribution in the body. In this study, we evaluate the expressions of BCL2 and BAX genes in T47D treated with cisplatin and cisplatin nanoparticles, which can result in a new approach to breast cancer therapy.
Methods: In this study, we treated T47D cells with different concentrations of cisplatin and cisplatin nanoparticles at 48 h. The IC50 was determined. We extracted RNA by using RNX solution, after which cDNA was synthesized. The precise primers for the BCL2, BAX and TBP genes were designed by specific software. The quantity of BCL2 and BAX gene expression compared to TBP gene (reference gene) was analyzed using real-time PCR.
Results: BCL2 and BAX gene expression levels in T47D cells treated by cisplatin were 0.7 (BCL2) and 1.48 (BAX); in T47D cells treated with cisplatin-loaded nanoparticles, the gene expressions were 0.03 (BCL2) and 2.41 (BAX).
Conclusion: In this study, the results have shown that cisplatin-loaded nanoparticles are effective anticancer agents. Cisplatin nanoparticles induce apoptosis in human breast cancer cell lines. We have shown that cisplatin nanoparticles strongly increased cytotoxicity in comparison to the free drug in the T47D cell line.

Keywords

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Pathobiology Reserach
Volume 15, Issue 4
February 2013
Pages 75-87